Research activity information

• Nov. 2022 兵庫県科学賞

  内匠透


• Oct. 2022 神戸大学学長表彰

  内匠透


• Oct. 2021 神戸大学学長表彰

  内匠透


• Oct. 2020 神戸大学学長表彰

  内匠透


• Mar. 2017 Terumo Foundatuon Prize

  Toru Takumi


• 2012 HFSP Research Award

  Toru Takumi


• Nov. 2009 ベルツ賞

  内匠透


• Jan. 2000 神戸大学医学部優秀論文賞

  内匠透


• 1991 HFSP, Long-term fellowship

  Toru Takumi

• Reorganization of Social Representations in the Insular Cortex During Conspecific Familiarization and Discrimination.

  Masaaki Sato, Eric T N Overton, Shuhei Fujima, Toru Takumi

  The familiarity of socially interacting peers markedly impacts behavior. However, the neuronal representations that distinguish familiar from novel conspecifics within the social brain network are not fully understood. Following our previous findings that neurons in the agranular insular cortex represent ongoing social interactions, we monitored the activity of neurons in the agranular insular cortex using microendoscopic calcium imaging in mice during social recognition memory and linear chamber social discrimination tasks. In the social recognition memory task, repeated interactions with the same target activated largely nonoverlapping cells during each session. The fraction of cells associated with social investigation (hereafter called social cells) decreased as the subject repeatedly interacted with the same target, whereas the substitution of a second novel target and subsequent exchange with the first familiar target recruited more new social cells. In the linear chamber social discrimination task, adding a novel target transiently increased the number of cells responding to both targets, followed by an eventual increase in the number of cells responding to the novel target. These results demonstrate that social cell ensembles in the agranular insular cortex decrease in size while changing their participating neurons during conspecific familiarization. They also rapidly reorganize at the single-cell level to represent interactions with novel peers rather than familiar peers during conspecific discrimination.

  Jul. 2025, The European journal of neuroscience, 62(2) (2), e70190, English, International magazine

  Scientific journal


• Editorial overview: Mechanisms underlying neurodevelopmental disorders.

  M Chiara Manzini, Toru Takumi

  Jun. 2025, Current opinion in neurobiology, 93, 103043 - 103043, English, International magazine


• ESC models of autism with copy-number variations reveal cell-type-specific translational vulnerability.

  Jun Nomura, Amila Zuko, Keiko Kishimoto, Hiroaki Mutsumine, Hiroko Maegawa, Kazumi Fukatsu, Yoshiko Nomura, Xiaoxi Liu, Nobuhiro Nakai, Eiki Takahashi, Tsukasa Kouno, Jay W Shin, Toru Takumi

  Human genetics has identified numerous copy-number variations (CNVs) associated with autism spectrum disorders (ASDs). However, the lack of standardized biological resources impedes understanding of the cell-type-specific common features of ASD. Here, we establish a biological resource including 63 genetically modified mouse embryonic stem cell (ESC) lines as genetic models of ASD. We perform neural differentiation using 12 representative cell lines, and their comprehensive analyses, including single-cell RNA sequencing, uncover cell-type-specific susceptible pathways. Moreover, we find that a common phenotype in glutamatergic and GABAergic neurons is reduced expression of Upf3b, a core member of the translational termination and nonsense-mediated decay (NMD). This finding emphasizes that the dysfunction of translational machinery in the developing neurons can be a possible target of early intervention for ASD. This ESC model bank becomes an invaluable resource for studies in vitro and in vivo of ASD or other neuropsychiatric disorders.

  Jun. 2025, Cell genomics, 5(6) (6), 100877 - 100877, English, International magazine

  Scientific journal


• Duplication of the autism-related gene Chd8 leads to behavioral hyperactivity and neurodevelopmental defects in mice.

  Atsuki Kawamura, Kazuki Fujii, Kota Tamada, Yoshifumi Abe, Kenta Nitahara, Tomoya Iwasaki, Sho Yagishita, Kenji F Tanaka, Toru Takumi, Keizo Takao, Masaaki Nishiyama

  Mutations in the gene encoding chromodomain helicase DNA-binding protein 8 (CHD8) are strongly associated with autism spectrum disorder (ASD). Although duplications of the chromosomal locus including CHD8 have also been detected in individuals with neurodevelopmental disorders, the contribution of CHD8 duplication to clinical phenotypes and the underlying mechanisms have remained unknown. Here we show that Chd8 knock-in (KI) mice that overexpress CHD8 as a model of human CHD8 duplication manifest growth retardation, microcephaly, impaired neuronal differentiation, and behavioral abnormalities including hyperactivity and reduced anxiety-like behavior. Chd8 overexpression affects the transcription and chromatin accessibility of genes related to neurogenesis, with these changes being associated with aberrant binding of CHD8 to enhancer regions. Furthermore, pharmacological intervention partially ameliorates the hyperactivity of Chd8 KI mice. Our results thus indicate that Chd8 KI mice recapitulate key features of CHD8 duplication syndrome in humans, providing insight into pathogenic mechanisms underlying neurodevelopmental disorders.

  May 2025, Nature communications, 16(1) (1), 4641 - 4641, English, International magazine

  Scientific journal


• Neurodevelopmental impact of CNV models in ASD: Recent advances and future directions.

  Kota Tamada, Toru Takumi

  Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by social communication impairments and restricted, repetitive behaviors. ASD exhibits a strong genetic basis, with rare and common genetic variants contributing to its etiology. Copy number variations (CNVs), deletions or duplications of chromosomal segments, have emerged as key contributors to ASD risk. Rare CNVs often demonstrate large effect sizes and can directly cause ASD, while common variants collectively exert subtle influences. Recent advances have identified numerous ASD-associated CNVs, including recurrent loci such as 1q21.1, 2p16.3, 7q11.23, 15q11.2, 15q11-q13, 16p11.2 and 22q11.2. Mouse models carrying these CNVs have provided profound insights into the underlying neurobiological mechanisms. Recent studies integrating transcriptomic, proteomic, and functional imaging approaches have revealed alterations in synaptic function, neuronal differentiation, myelination, metabolic pathways, and circuit connectivity. Notably, investigations leveraging conditional knockout models, high magnetic field MRI, and single-cell analyses highlight disruptions in excitatory-inhibitory balance, white matter integrity, and dynamic gene regulatory networks. Parallel human-based approaches, including iPSC-derived neurons, cerebral organoids, and large-scale single-nucleus sequencing, are combined with animal model data. These integrative strategies promise to refine our understanding of ASD's genetic architecture, bridging the gap between fundamental discoveries in model organisms and clinically relevant biomarkers, subtypes, and therapeutic targets in humans. This review summarizes key findings from recent CNV mouse model studies and highlights emerging technologies applied to human ASD samples. Finally, we outline prospects for translating findings from mouse studies to humans. By illuminating both unique and convergent genetic mechanisms, these advances offer a critical framework for unraveling etiological complexity in ASD.

  Mar. 2025, Current opinion in neurobiology, 92, 103001 - 103001, English, International magazine

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Correlations of brain structure with the social behavior of 15q11-13 duplication mice, an animal model of autism.

  Zhilei Zhao, Naohiro Okada, Sho Yagishita, Noriaki Yahata, Nobuhiro Nitta, Sayaka Shibata, Yoshifumi Abe, Susumu Morita, Eureka Kumagai, Kenji F Tanaka, Tetsuya Suhara, Toru Takumi, Kiyoto Kasai, Seiichiro Jinde

  Duplication of chromosome 15q11-13 has been reported to be one of the most frequent cytogenetic copy number variations in autism spectrum disorder (ASD), and a mouse model of paternal 15q11-13 duplication was generated, termed 15q dup mice. While previous studies have replicated some of the behavioral and brain structural phenotypes of ASD separately, the relationship between brain structure and behavior has rarely been examined. In this study, we performed behavioral experiments related to anxiety and social behaviors and magnetic resonance imaging (MRI) using the same set of 15q dup and wild-type mice. 15q dup mice showed increased anxiety and a tendency toward alterations in social behaviors, as reported previously, as well as variability in terms of sociability. MRI analysis revealed that a lower sociability index was correlated with a smaller gray matter volume in the right medial entorhinal cortex. These results may help to understand how variability in behavioral phenotypes of ASD arises even in individuals with the same genetic background and to determine the individual differences in neurodevelopmental trajectory correlated with specific brain structures that underlie these phenotypes.

  Aug. 2024, Neuroscience research, 209, 42 - 49, English, International magazine

  Scientific journal


• Sleep EEG signatures in mouse models of 15q11.2-13.1 duplication (Dup15q) syndrome.

  Vidya Saravanapandian, Melika Madani, India Nichols, Scott Vincent, Mary Dover, Dante Dikeman, Benjamin D Philpot, Toru Takumi, Christopher S Colwell, Shafali Jeste, Ketema N Paul, Peyman Golshani

  BACKGROUND: Sleep disturbances are a prevalent and complex comorbidity in neurodevelopmental disorders (NDDs). Dup15q syndrome (duplications of 15q11.2-13.1) is a genetic disorder highly penetrant for NDDs such as autism and intellectual disability and it is frequently accompanied by significant disruptions in sleep patterns. The 15q critical region harbors genes crucial for brain development, notably UBE3A and a cluster of gamma-aminobutyric acid type A receptor (GABAAR) genes. We previously described an electrophysiological biomarker of the syndrome, marked by heightened beta oscillations (12-30 Hz) in individuals with Dup15q syndrome, akin to electroencephalogram (EEG) alterations induced by allosteric modulation of GABAARs. Those with Dup15q syndrome exhibited increased beta oscillations during the awake resting state and during sleep, and they showed profoundly abnormal NREM sleep. This study aims to assess the translational validity of these EEG signatures and to delve into their neurobiological underpinnings by quantifying sleep physiology in chromosome-engineered mice with maternal (matDp/ + mice) or paternal (patDp/ + mice) inheritance of the full 15q11.2-13.1-equivalent duplication, and mice with duplication of just the UBE3A gene (Ube3a overexpression mice; Ube3a OE mice) and comparing the sleep metrics with their respective wildtype (WT) littermate controls. METHODS: We collected 48-h EEG/EMG recordings from 35 (23 male, 12 female) 12-24-week-old matDp/ + , patDp/ + , Ube3a OE mice, and their WT littermate controls. We quantified baseline sleep, sleep fragmentation, spectral power dynamics during sleep states, and recovery following sleep deprivation. Within each group, distinctions between Dup15q mutant mice and WT littermate controls were evaluated using analysis of variance (ANOVA) and student's t-test. The impact of genotype and time was discerned through repeated measures ANOVA, and significance was established at p < 0.05. RESULTS: Our study revealed that across brain states, matDp/ + mice mirrored the elevated beta oscillation phenotype observed in clinical EEGs from individuals with Dup15q syndrome. Time to sleep onset after light onset was significantly reduced in matDp/ + and Ube3a OE mice. However, NREM sleep between Dup15q mutant and WT littermate mice remained unaltered, suggesting a divergence from the clinical presentation in humans. Additionally, while increased beta oscillations persisted in matDp/ + mice after 6-h of sleep deprivation, recovery NREM sleep remained unaltered in all groups, thus suggesting that these mice exhibit resilience in the fundamental processes governing sleep-wake regulation. CONCLUSIONS: Quantification of mechanistic and translatable EEG biomarkers is essential for advancing our understanding of NDDs and their underlying pathophysiology. Our study of sleep physiology in the Dup15q mice underscores that the beta EEG biomarker has strong translational validity, thus opening the door for pre-clinical studies of putative drug targets, using the biomarker as a translational measure of drug-target engagement. The unaltered NREM sleep may be due to inherent differences in neurobiology between mice and humans. These nuanced distinctions highlight the complexity of sleep disruptions in Dup15q syndrome and emphasize the need for a comprehensive understanding that encompasses both shared and distinct features between murine models and clinical populations.

  Jul. 2024, Journal of neurodevelopmental disorders, 16(1) (1), 39 - 39, English, International magazine

  Scientific journal


• Alteration of synaptic protein composition during developmental synapse maturation.

  Takeshi Kaizuka, Toru Takumi

  The postsynaptic density (PSD) is a collection of specialized proteins assembled beneath the postsynaptic membrane of dendritic spines. The PSD proteome comprises ~1000 proteins, including neurotransmitter receptors, scaffolding proteins and signalling enzymes. Many of these proteins have essential roles in synaptic function and plasticity. During brain development, changes are observed in synapse density and in the stability and shape of spines, reflecting the underlying molecular maturation of synapses. Synaptic protein composition changes in terms of protein abundance and the assembly of protein complexes, supercomplexes and the physical organization of the PSD. Here, we summarize the developmental alterations of postsynaptic protein composition during synapse maturation. We describe major PSD proteins involved in postsynaptic signalling that regulates synaptic plasticity and discuss the effect of altered expression of these proteins during development. We consider the abnormality of synaptic profiles and synaptic protein composition in the brain in neurodevelopmental disorders such as autism spectrum disorders. We also explain differences in synapse development between rodents and primates in terms of synaptic profiles and protein composition. Finally, we introduce recent findings related to synaptic diversity and nanoarchitecture and discuss their impact on future research. Synaptic protein composition can be considered a major determinant and marker of synapse maturation in normality and disease.

  Jun. 2024, The European journal of neuroscience, 59(11) (11), 2894 - 2914, English, International magazine

  Scientific journal


• Remodeling of the postsynaptic proteome in male mice and marmosets during synapse development

  Takeshi Kaizuka, Takehiro Suzuki, Noriyuki Kishi, Kota Tamada, Manfred W. Kilimann, Takehiko Ueyama, Masahiko Watanabe, Tomomi Shimogori, Hideyuki Okano, Naoshi Dohmae, Toru Takumi

  Abstract Postsynaptic proteins play crucial roles in synaptic function and plasticity. During brain development, alterations in synaptic number, shape, and stability occur, known as synapse maturation. However, the postsynaptic protein composition changes during development are not fully understood. Here, we show the trajectory of the postsynaptic proteome in developing male mice and common marmosets. Proteomic analysis of mice at 2, 3, 6, and 12 weeks of age shows that proteins involved in synaptogenesis are differentially expressed during this period. Analysis of published transcriptome datasets shows that the changes in postsynaptic protein composition in the mouse brain after 2 weeks of age correlate with gene expression changes. Proteomic analysis of marmosets at 0, 2, 3, 6, and 24 months of age show that the changes in the marmoset brain can be categorized into two parts: the first 2 months and after that. The changes observed in the first 2 months are similar to those in the mouse brain between 2 and 12 weeks of age. The changes observed in marmoset after 2 months old include differential expression of synaptogenesis-related molecules, which hardly overlap with that in mice. Our results provide a comprehensive proteomic resource that underlies developmental synapse maturation in rodents and primates.

  Springer Science and Business Media LLC, Mar. 2024, Nature Communications, 15(1) (1)

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Large-scale animal model study uncovers altered brain pH and lactate levels as a transdiagnostic endophenotype of neuropsychiatric disorders involving cognitive impairment.

  Hideo Hagihara, Hirotaka Shoji, Satoko Hattori, Giovanni Sala, Yoshihiro Takamiya, Mika Tanaka, Masafumi Ihara, Mihiro Shibutani, Izuho Hatada, Kei Hori, Mikio Hoshino, Akito Nakao, Yasuo Mori, Shigeo Okabe, Masayuki Matsushita, Anja Urbach, Yuta Katayama, Akinobu Matsumoto, Keiichi I Nakayama, Shota Katori, Takuya Sato, Takuji Iwasato, Haruko Nakamura, Yoshio Goshima, Matthieu Raveau, Tetsuya Tatsukawa, Kazuhiro Yamakawa, Noriko Takahashi, Haruo Kasai, Johji Inazawa, Ikuo Nobuhisa, Tetsushi Kagawa, Tetsuya Taga, Mohamed Darwish, Hirofumi Nishizono, Keizo Takao, Kiran Sapkota, Kazutoshi Nakazawa, Tsuyoshi Takagi, Haruki Fujisawa, Yoshihisa Sugimura, Kyosuke Yamanishi, Lakshmi Rajagopal, Nanette Deneen Hannah, Herbert Y Meltzer, Tohru Yamamoto, Shuji Wakatsuki, Toshiyuki Araki, Katsuhiko Tabuchi, Tadahiro Numakawa, Hiroshi Kunugi, Freesia L Huang, Atsuko Hayata-Takano, Hitoshi Hashimoto, Kota Tamada, Toru Takumi, Takaoki Kasahara, Tadafumi Kato, Isabella A Graef, Gerald R Crabtree, Nozomi Asaoka, Hikari Hatakama, Shuji Kaneko, Takao Kohno, Mitsuharu Hattori, Yoshio Hoshiba, Ryuhei Miyake, Kisho Obi-Nagata, Akiko Hayashi-Takagi, Léa J Becker, Ipek Yalcin, Yoko Hagino, Hiroko Kotajima-Murakami, Yuki Moriya, Kazutaka Ikeda, Hyopil Kim, Bong-Kiun Kaang, Hikari Otabi, Yuta Yoshida, Atsushi Toyoda, Noboru H Komiyama, Seth G N Grant, Michiru Ida-Eto, Masaaki Narita, Ken-Ichi Matsumoto, Emiko Okuda-Ashitaka, Iori Ohmori, Tadayuki Shimada, Kanato Yamagata, Hiroshi Ageta, Kunihiro Tsuchida, Kaoru Inokuchi, Takayuki Sassa, Akio Kihara, Motoaki Fukasawa, Nobuteru Usuda, Tayo Katano, Teruyuki Tanaka, Yoshihiro Yoshihara, Michihiro Igarashi, Takashi Hayashi, Kaori Ishikawa, Satoshi Yamamoto, Naoya Nishimura, Kazuto Nakada, Shinji Hirotsune, Kiyoshi Egawa, Kazuma Higashisaka, Yasuo Tsutsumi, Shoko Nishihara, Noriyuki Sugo, Takeshi Yagi, Naoto Ueno, Tomomi Yamamoto, Yoshihiro Kubo, Rie Ohashi, Nobuyuki Shiina, Kimiko Shimizu, Sayaka Higo-Yamamoto, Katsutaka Oishi, Hisashi Mori, Tamio Furuse, Masaru Tamura, Hisashi Shirakawa, Daiki X Sato, Yukiko U Inoue, Takayoshi Inoue, Yuriko Komine, Tetsuo Yamamori, Kenji Sakimura, Tsuyoshi Miyakawa

  Increased levels of lactate, an end-product of glycolysis, have been proposed as a potential surrogate marker for metabolic changes during neuronal excitation. These changes in lactate levels can result in decreased brain pH, which has been implicated in patients with various neuropsychiatric disorders. We previously demonstrated that such alterations are commonly observed in five mouse models of schizophrenia, bipolar disorder, and autism, suggesting a shared endophenotype among these disorders rather than mere artifacts due to medications or agonal state. However, there is still limited research on this phenomenon in animal models, leaving its generality across other disease animal models uncertain. Moreover, the association between changes in brain lactate levels and specific behavioral abnormalities remains unclear. To address these gaps, the International Brain pH Project Consortium investigated brain pH and lactate levels in 109 strains/conditions of 2294 animals with genetic and other experimental manipulations relevant to neuropsychiatric disorders. Systematic analysis revealed that decreased brain pH and increased lactate levels were common features observed in multiple models of depression, epilepsy, Alzheimer's disease, and some additional schizophrenia models. While certain autism models also exhibited decreased pH and increased lactate levels, others showed the opposite pattern, potentially reflecting subpopulations within the autism spectrum. Furthermore, utilizing large-scale behavioral test battery, a multivariate cross-validated prediction analysis demonstrated that poor working memory performance was predominantly associated with increased brain lactate levels. Importantly, this association was confirmed in an independent cohort of animal models. Collectively, these findings suggest that altered brain pH and lactate levels, which could be attributed to dysregulated excitation/inhibition balance, may serve as transdiagnostic endophenotypes of debilitating neuropsychiatric disorders characterized by cognitive impairment, irrespective of their beneficial or detrimental nature.

  Mar. 2024, eLife, 12, English, International magazine

  Scientific journal


• Transcriptomic dysregulation and autistic-like behaviors in Kmt2c haploinsufficient mice rescued by an LSD1 inhibitor.

  Takumi Nakamura, Toru Yoshihara, Chiharu Tanegashima, Mitsutaka Kadota, Yuki Kobayashi, Kurara Honda, Mizuho Ishiwata, Junko Ueda, Tomonori Hara, Moe Nakanishi, Toru Takumi, Shigeyoshi Itohara, Shigehiro Kuraku, Masahide Asano, Takaoki Kasahara, Kazuo Nakajima, Takashi Tsuboi, Atsushi Takata, Tadafumi Kato

  Recent studies have consistently demonstrated that the regulation of chromatin and gene transcription plays a pivotal role in the pathogenesis of neurodevelopmental disorders. Among many genes involved in these pathways, KMT2C, encoding one of the six known histone H3 lysine 4 (H3K4) methyltransferases in humans and rodents, was identified as a gene whose heterozygous loss-of-function variants are causally associated with autism spectrum disorder (ASD) and the Kleefstra syndrome phenotypic spectrum. However, little is known about how KMT2C haploinsufficiency causes neurodevelopmental deficits and how these conditions can be treated. To address this, we developed and analyzed genetically engineered mice with a heterozygous frameshift mutation of Kmt2c (Kmt2c+/fs mice) as a disease model with high etiological validity. In a series of behavioral analyses, the mutant mice exhibit autistic-like behaviors such as impairments in sociality, flexibility, and working memory, demonstrating their face validity as an ASD model. To investigate the molecular basis of the observed abnormalities, we performed a transcriptomic analysis of their bulk adult brains and found that ASD risk genes were specifically enriched in the upregulated differentially expressed genes (DEGs), whereas KMT2C peaks detected by ChIP-seq were significantly co-localized with the downregulated genes, suggesting an important role of putative indirect effects of Kmt2c haploinsufficiency. We further performed single-cell RNA sequencing of newborn mouse brains to obtain cell type-resolved insights at an earlier stage. By integrating findings from ASD exome sequencing, genome-wide association, and postmortem brain studies to characterize DEGs in each cell cluster, we found strong ASD-associated transcriptomic changes in radial glia and immature neurons with no obvious bias toward upregulated or downregulated DEGs. On the other hand, there was no significant gross change in the cellular composition. Lastly, we explored potential therapeutic agents and demonstrate that vafidemstat, a lysine-specific histone demethylase 1 (LSD1) inhibitor that was effective in other models of neuropsychiatric/neurodevelopmental disorders, ameliorates impairments in sociality but not working memory in adult Kmt2c+/fs mice. Intriguingly, the administration of vafidemstat was shown to alter the vast majority of DEGs in the direction to normalize the transcriptomic abnormalities in the mutant mice (94.3 and 82.5% of the significant upregulated and downregulated DEGs, respectively, P < 2.2 × 10-16, binomial test), which could be the molecular mechanism underlying the behavioral rescuing. In summary, our study expands the repertoire of ASD models with high etiological and face validity, elucidates the cell-type resolved molecular alterations due to Kmt2c haploinsufficiency, and demonstrates the efficacy of an LSD1 inhibitor that might be generalizable to multiple categories of psychiatric disorders along with a better understanding of its presumed mechanisms of action.

  Mar. 2024, Molecular psychiatry, English, International magazine

  Scientific journal


• End-to-end deep learning approach to mouse behavior classification from cortex-wide calcium imaging.

  Takehiro Ajioka, Nobuhiro Nakai, Okito Yamashita, Toru Takumi

  Deep learning is a powerful tool for neural decoding, broadly applied to systems neuroscience and clinical studies. Interpretable and transparent models that can explain neural decoding for intended behaviors are crucial to identifying essential features of deep learning decoders in brain activity. In this study, we examine the performance of deep learning to classify mouse behavioral states from mesoscopic cortex-wide calcium imaging data. Our convolutional neural network (CNN)-based end-to-end decoder combined with recurrent neural network (RNN) classifies the behavioral states with high accuracy and robustness to individual differences on temporal scales of sub-seconds. Using the CNN-RNN decoder, we identify that the forelimb and hindlimb areas in the somatosensory cortex significantly contribute to behavioral classification. Our findings imply that the end-to-end approach has the potential to be an interpretable deep learning method with unbiased visualization of critical brain regions.

  Mar. 2024, PLoS computational biology, 20(3) (3), e1011074, English, International magazine

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• FAM81A is a postsynaptic protein that regulates the condensation of postsynaptic proteins via liquid-liquid phase separation.

  Takeshi Kaizuka, Taisei Hirouchi, Takeo Saneyoshi, Toshihiko Shirafuji, Mark O Collins, Seth G N Grant, Yasunori Hayashi, Toru Takumi

  Proteome analyses of the postsynaptic density (PSD), a proteinaceous specialization beneath the postsynaptic membrane of excitatory synapses, have identified several thousands of proteins. While proteins with predictable functions have been well studied, functionally uncharacterized proteins are mostly overlooked. In this study, we conducted a comprehensive meta-analysis of 35 PSD proteome datasets, encompassing a total of 5,869 proteins. Employing a ranking methodology, we identified 97 proteins that remain inadequately characterized. From this selection, we focused our detailed analysis on the highest-ranked protein, FAM81A. FAM81A interacts with PSD proteins, including PSD-95, SynGAP, and NMDA receptors, and promotes liquid-liquid phase separation of those proteins in cultured cells or in vitro. Down-regulation of FAM81A in cultured neurons causes a decrease in the size of PSD-95 puncta and the frequency of neuronal firing. Our findings suggest that FAM81A plays a crucial role in facilitating the interaction and assembly of proteins within the PSD, and its presence is important for maintaining normal synaptic function. Additionally, our methodology underscores the necessity for further characterization of numerous synaptic proteins that still lack comprehensive understanding.

  Mar. 2024, PLoS biology, 22(3) (3), e3002006, English, International magazine

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• The blues and rhythm.

  Kiyomichi Imamura, Ayaka Bota, Toshihiko Shirafuji, Toru Takumi

  Most organisms, including humans, show daily rhythms in many aspects of physiology and behavior, and abnormalities in the rhythms are potential risk factors for various diseases. Mood disorders such as depression are no exception. Accumulating evidence suggests strong associations between circadian disturbances and the development of depression. Numerous studies have shown that interventions to circadian rhythms trigger depression-like phenotypes in human cases and animal models. Conversely, mood changes can affect circadian rhythms as symptoms of depression. Our preliminary data suggest that the phosphorylation signal pathway of the clock protein may act as a common pathway for mood and clock regulation. We hypothesize that mood regulation and circadian rhythms may influence each other and may share a common regulatory mechanism. This review provides an overview of circadian disturbances in animal models and human patients with depression.

  Nov. 2023, Neuroscience research, English, International magazine

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• The East Asian–Specific Risk Genes in Autism Spectrum Disorder

  Tamada, K., Takumi, T.

  Nov. 2023, Biological Psychiatry, 94(10) (10), 762 - 764, English

  [Invited]

  Scientific journal


• Social circuits and their dysfunction in autism spectrum disorder.

  Masaaki Sato, Nobuhiro Nakai, Shuhei Fujima, Katrina Y Choe, Toru Takumi

  Social behaviors, how individuals act cooperatively and competitively with conspecifics, are widely seen across species. Rodents display various social behaviors, and many different behavioral paradigms have been used for investigating their neural circuit bases. Social behavior is highly vulnerable to brain network dysfunction caused by neurological and neuropsychiatric conditions such as autism spectrum disorders (ASDs). Studying mouse models of ASD provides a promising avenue toward elucidating mechanisms of abnormal social behavior and potential therapeutic targets for treatment. In this review, we outline recent progress and key findings on neural circuit mechanisms underlying social behavior, with particular emphasis on rodent studies that monitor and manipulate the activity of specific circuits using modern systems neuroscience approaches. Social behavior is mediated by a distributed brain-wide network among major cortical (e.g., medial prefrontal cortex (mPFC), anterior cingulate cortex, and insular cortex (IC)) and subcortical (e.g., nucleus accumbens, basolateral amygdala (BLA), and ventral tegmental area) structures, influenced by multiple neuromodulatory systems (e.g., oxytocin, dopamine, and serotonin). We particularly draw special attention to IC as a unique cortical area that mediates multisensory integration, encoding of ongoing social interaction, social decision-making, emotion, and empathy. Additionally, a synthesis of studies investigating ASD mouse models demonstrates that dysfunctions in mPFC-BLA circuitry and neuromodulation are prominent. Pharmacological rescues by local or systemic (e.g., oral) administration of various drugs have provided valuable clues for developing new therapeutic agents for ASD. Future efforts and technological advances will push forward the next frontiers in this field, such as the elucidation of brain-wide network activity and inter-brain neural dynamics during real and virtual social interactions, and the establishment of circuit-based therapy for disorders affecting social functions.

  Aug. 2023, Molecular psychiatry, English, International magazine

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Germ-cell-specific transcriptome analysis illuminates the chromatin and ubiquitin pathway in autism spectrum disorders.

  Sawako Furukawa, Jun Nomura, Hiroaki Hanafusa, Hiroko Maegawa, Toru Takumi

  Accumulating epidemiological studies have suggested a positive association between advanced paternal age at conception and the increased risk of neurodevelopmental outcomes such as autism spectrum disorder (ASD) in their children. Recent biological studies using human sperm have identified increased de novo mutations in aged fathers, and hyper- or hypomethylation has been identified in the sperm from aged rodents. Dysregulation of DNA methylation in sperm may explain the transgenerational effects on the pathogenesis of ASD. However, compared to these epigenetic changes in the sperm of aged males, the effects of inherited predisposition from germ cells are largely unknown. Here, we use single-cell transcriptome data sets from 13 cell lines, including 12 ASD-associated CNVs models and control, that are performed neural differentiation from mouse embryonic stem cells. This study performed comprehensive bioinformatic analyses such as gene ontology (GO), network, pathway, and upstream regulator analyses. Through these analyses, we identify several susceptible pathways, such as chromatin and ubiquitin, in addition to translational and oxidative phosphorylation. Our results suggest that dysregulation of epigenetic chromosome remodeling and ubiquitin-proteasome pathway in the germ cell is a possible modulator for subsequent differentiated cells, sperm, and egg, as a risk factor for the neurodevelopmental disorder.

  Jun. 2023, Autism research : official journal of the International Society for Autism Research, 16(6) (6), 1101 - 1110, English, International magazine

  Scientific journal


• Virtual reality-based real-time imaging reveals abnormal cortical dynamics during behavioral transitions in a mouse model of autism.

  Nobuhiro Nakai, Masaaki Sato, Okito Yamashita, Yukiko Sekine, Xiaochen Fu, Junichi Nakai, Andrew Zalesky, Toru Takumi

  Functional connectivity (FC) can provide insight into cortical circuit dysfunction in neuropsychiatric disorders. However, dynamic changes in FC related to locomotion with sensory feedback remain to be elucidated. To investigate FC dynamics in locomoting mice, we develop mesoscopic Ca2+ imaging with a virtual reality (VR) environment. We find rapid reorganization of cortical FC in response to changing behavioral states. By using machine learning classification, behavioral states are accurately decoded. We then use our VR-based imaging system to study cortical FC in a mouse model of autism and find that locomotion states are associated with altered FC dynamics. Furthermore, we identify FC patterns involving the motor area as the most distinguishing features of the autism mice from wild-type mice during behavioral transitions, which might correlate with motor clumsiness in individuals with autism. Our VR-based real-time imaging system provides crucial information to understand FC dynamics linked to behavioral abnormality of neuropsychiatric disorders.

  Mar. 2023, Cell reports, 112258 - 112258, English, International magazine

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• An old model with new insights: endogenous retroviruses drive the evolvement toward ASD susceptibility and hijack transcription machinery during development.

  Chia-Wen Lin, Jacob Ellegood, Kota Tamada, Ikuo Miura, Mikiko Konda, Kozue Takeshita, Koji Atarashi, Jason P Lerch, Shigeharu Wakana, Thomas J McHugh, Toru Takumi

  The BTBR T+Itpr3tf/J (BTBR/J) strain is one of the most valid models of idiopathic autism, serving as a potent forward genetics tool to dissect the complexity of autism. We found that a sister strain with an intact corpus callosum, BTBR TF/ArtRbrc (BTBR/R), showed more prominent autism core symptoms but moderate ultrasonic communication/normal hippocampus-dependent memory, which may mimic autism in the high functioning spectrum. Intriguingly, disturbed epigenetic silencing mechanism leads to hyperactive endogenous retrovirus (ERV), a mobile genetic element of ancient retroviral infection, which increases de novo copy number variation (CNV) formation in the two BTBR strains. This feature makes the BTBR strain a still evolving multiple-loci model toward higher ASD susceptibility. Furthermore, active ERV, analogous to virus infection, evades the integrated stress response (ISR) of host defense and hijacks the transcriptional machinery during embryonic development in the BTBR strains. These results suggest dual roles of ERV in the pathogenesis of ASD, driving host genome evolution at a long-term scale and managing cellular pathways in response to viral infection, which has immediate effects on embryonic development. The wild-type Draxin expression in BTBR/R also makes this substrain a more precise model to investigate the core etiology of autism without the interference of impaired forebrain bundles as in BTBR/J.

  Mar. 2023, Molecular psychiatry, English, International magazine

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Analysis of cortical network dynamics in the behavioral state of a mouse model of autism

  Nakai, N., Takumi, T.

  2023, Folia Pharmacologica Japonica, 158(2) (2)

  Scientific journal


• Increased self-triggered vocalizations in an epidermal growth factor-induced rat model for schizophrenia

  Itaru Narihara, Hanako Yokoyama, Hisaaki Namba, Hidekazu Sotoyama, Hiroyoshi Inaba, Eiko Kitayama, Kota Tamada, Toru Takumi, Hiroyuki Nawa

  Abstract Rats elicit two types of ultrasonic vocalizations (USVs), positive (30–80 kHz; high pitch) and negative (10–30 kHz; low pitch) voices. As patients with schizophrenia often exhibit soliloquy-like symptoms, we explored whether an animal model for schizophrenia is similarly characterized by such self-triggered vocalizations. We prepared the animal model by administering an inflammatory cytokine, epidermal growth factor (EGF), to rat neonates, which later develop behavioral and electroencephalographic deficits relevant to schizophrenia. EGF model rats and controls at young (8–10 weeks old) and mature (12–14 weeks old) adult stages were subjected to acclimation, female pairing, and vocalization sessions. In acclimation sessions, low pitch USVs at the mature adult stage were more frequent in EGF model rats than in controls. In the vocalization session, the occurrences of low pitch self-triggered USVs were higher in EGF model rats in both age groups, although this group difference was eliminated by their risperidone treatment. Unlike conventional negative USVs of rats, however, the present low pitch self-triggered USVs had short durations of 10–30 ms. These results suggest the potential that self-triggered vocalization might serve as a translatable pathological trait of schizophrenia to animal models.

  Springer Science and Business Media LLC, Dec. 2022, Scientific Reports, 12(1) (1)

  Scientific journal


• Mood phenotypes in rodent models with circadian disturbances.

  Kiyomichi Imamura, Toru Takumi

  Many physiological functions with approximately 24-h rhythmicity (circadian rhythms) are generated by an internal time-measuring system of the circadian clock. While sleep/wake cycles, feeding patterns, and body temperature are the most widely known physiological functions under the regulation of the circadian clock, physiological regulation by the circadian clock extends to higher brain functions. Accumulating evidence suggests strong associations between the circadian clock and mood disorders such as depression, but the underlying mechanisms of the functional relationship between them are obscure. This review overviews rodent models with disrupted circadian rhythms on depression-related responses. The animal models with circadian disturbances (by clock gene mutations and artifactual interventions) will help understand the causal link between the circadian clock and depression.

  Nov. 2022, Neurobiology of sleep and circadian rhythms, 13, 100083 - 100083, English, International magazine

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Species-specific formation of paraspeckles in intestinal epithelium revealed by characterization of NEAT1 in naked mole-rat.

  Akihiro Yamada, Hikaru Toya, Mayuko Tanahashi, Misuzu Kurihara, Mari Mito, Shintaro Iwasaki, Satoshi Kurosaka, Toru Takumi, Archa Fox, Yoshimi Kawamura, Kyoko Miura, Shinichi Nakagawa

  Abstract Paraspeckles are mammalian-specific nuclear bodies built on the long noncoding RNA NEAT1_2. The molecular mechanisms of paraspeckle formation have been mainly studied using human or mouse cells, and it is not known if the same molecular components are involved in the formation of paraspeckles in other mammalian species. We thus investigated the expression pattern of NEAT1_2 in naked mole-rats (nNEAT1_2), which exhibit extreme longevity and lower susceptibility to cancer. In the intestine, nNEAT1_2 is widely expressed along the entire intestinal epithelium, which is different from the expression of mNeat1_2, that is restricted to the cells of the distal tip in mice. Notably, the expression of FUS, a FET family RNA binding protein, essential for the formation of paraspeckles both in humans and mice, was absent in the distal part of the intestinal epithelium in naked mole-rats. Instead, mRNAs of other FET family proteins EWSR1 and TAF15 were expressed in the distal region. Exogenous expression of these proteins in Fus-deficient murine embryonic fibroblast cells rescued the formation of paraspeckles. These observations suggest that nNEAT1_2 recruits different set of RNA binding proteins in a cell type-specific manner during the formation of paraspeckles in different organisms.

  Cold Spring Harbor Laboratory, Aug. 2022, RNA (New York, N.Y.), 28(8) (8), 1128 - 1143, English, International magazine

  Scientific journal


• A common epigenetic mechanism across different cellular origins underlies systemic immune dysregulation in an idiopathic autism mouse model.

  Chia-Wen Lin, Dian E Septyaningtrias, Hsu-Wen Chao, Mikiko Konda, Koji Atarashi, Kozue Takeshita, Kota Tamada, Jun Nomura, Yohei Sasagawa, Kaori Tanaka, Itoshi Nikaido, Kenya Honda, Thomas J McHugh, Toru Takumi

  Immune dysregulation plays a key role in the pathogenesis of autism. Changes occurring at the systemic level, from brain inflammation to disturbed innate/adaptive immune in the periphery, are frequently observed in patients with autism; however, the intrinsic mechanisms behind them remain elusive. We hypothesize a common etiology may lie in progenitors of different types underlying widespread immune dysregulation. By single-cell RNA sequencing (sc-RNA seq), we trace the developmental origins of immune dysregulation in a mouse model of idiopathic autism. It is found that both in aorta-gonad-mesonephros (AGM) and yolk sac (YS) progenitors, the dysregulation of HDAC1-mediated epigenetic machinery alters definitive hematopoiesis during embryogenesis and downregulates the expression of the AP-1 complex for microglia development. Subsequently, these changes result in the dysregulation of the immune system, leading to gut dysbiosis and hyperactive microglia in the brain. We further confirm that dysregulated immune profiles are associated with specific microbiota composition, which may serve as a biomarker to identify autism of immune-dysregulated subtypes. Our findings elucidate a shared mechanism for the origin of immune dysregulation from the brain to the gut in autism and provide new insight to dissecting the heterogeneity of autism, as well as the therapeutic potential of targeting immune-dysregulated autism subtypes.

  May 2022, Molecular psychiatry, 27(8) (8), 3343 - 3354, English, International magazine

  Scientific journal


• mTOR-AKT Signaling in Cellular Clock Resetting Triggered by Osmotic Stress.

  Hikari Yoshitane, Kiyomichi Imamura, Takenori Okubo, Yuta Otobe, Satoshi Kawakami, Shunsuke Ito, Toru Takumi, Kazuki Hattori, Isao Naguro, Hidenori Ichijo, Yoshitaka Fukada

  Aims: The circadian clock oscillates in a cell-autonomous manner with a period of ∼24 h, and the phase is regulated by various time cues such as light and temperature through multiple clock input pathways. We previously found that osmotic and oxidative stress strongly affected the circadian period and phase of cellular rhythms, and triple knockout of apoptosis signal-regulating kinase (ASK) family members, Ask1, Ask2, and Ask3, abolished the phase shift (clock resetting) induced by hyperosmotic pulse treatment. We aimed at exploring a key molecule(s) and signaling events in the clock input pathway dependent on ASK kinases. Results: The phase shift of the cellular clock induced by the hyperosmotic pulse treatment was significantly reduced by combined deficiencies of the clock(-related) genes, Dec1, Dec2, and E4 promoter-binding protein 4 (also known as Nfil3) (E4bp4). In addition, liquid chromatography mass/mass spectrometry (LC-MS/MS)-based proteomic analysis identified hyperosmotic pulse-induced phosphorylation of circadian locomotor output cycles caput (CLOCK) Ser845 in an AKT-dependent manner. We found that AKT kinase was phosphorylated at Ser473 (i.e., activated) in response to the hyperosmotic pulse experiments. Inhibition of mechanistic target of rapamycin (mTOR) kinase by Torin 1 treatment completely abolished the AKT activation, suppressed the phosphorylation of CLOCK Ser845, and blocked the clock resetting induced by the hyperosmotic pulse treatment. Innovation and Conclusions: We conclude that mTOR-AKT signaling is indispensable for the CLOCK Ser845 phosphorylation, which correlates with the clock resetting induced by the hyperosmotic pulse treatment. Immediate early induction of the clock(-related) genes and CLOCK carboxyl-terminal (C-terminal) region containing Ser845 also play important roles in the clock input pathway through redox-sensitive ASK kinases.

  Mar. 2022, Antioxidants & redox signaling, English, International magazine

  Scientific journal


• Sensing the Sounds of Silence: A Pilot Study on the Detection of Model Mice of Autism Spectrum Disorder from Ultrasonic Vocalisations.

  Kun Qian, Tomoya Koike, Kota Tamada, Toru Takumi, Bjorn W Schuller, Yoshiharu Yamamoto

  Studying the animal models of human neuropsychiatric disorders can facilitate the understanding of mechanisms of symptoms both physiologically and genetically. Previous studies have shown that ultrasonic vocalisations (USVs) of mice might be efficient markers to distinguish the wild type group and the model of autism spectrum disorder (mASD). Nevertheless, in-depth analysis of these 'silence' sounds by leveraging the power of advanced computer audition technologies (e. g., deep learning) is limited. To this end, we propose a pilot study on using a large-scale pre-trained audio neural network to extract high-level representations from the USVs of mice for the task on detection of mASD. Experiments have shown a best result reaching an unweighted average recall of 79.2 % for the binary classification task in a rigorous subject-independent scenario. To the best of our knowledge, this is the first time to analyse the sounds that cannot be heard by human beings for the detection of mASD mice. The novel findings can be significant to motivate future works with according means on studying animal models of human patients.

  Nov. 2021, Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Annual International Conference, 2021, 68 - 71, English, International magazine

  Scientific journal


• CHRONO and DEC1/DEC2 compensate for lack of CRY1/CRY2 in expression of coherent circadian rhythm but not in generation of circadian oscillation in the neonatal mouse SCN.

  Daisuke Ono, Ken-Ichi Honma, Christoph Schmal, Toru Takumi, Takeshi Kawamoto, Katsumi Fujimoto, Yukio Kato, Sato Honma

  Clock genes Cry1 and Cry2, inhibitory components of core molecular feedback loop, are regarded as critical molecules for the circadian rhythm generation in mammals. A double knockout of Cry1 and Cry2 abolishes the circadian behavioral rhythm in adult mice under constant darkness. However, robust circadian rhythms in PER2::LUC expression are detected in the cultured suprachiasmatic nucleus (SCN) of Cry1/Cry2 deficient neonatal mice and restored in adult SCN by co-culture with wild-type neonatal SCN. These findings led us to postulate the compensatory molecule(s) for Cry1/Cry2 deficiency in circadian rhythm generation. We examined the roles of Chrono and Dec1/Dec2 proteins, the suppressors of Per(s) transcription similar to CRY(s). Unexpectedly, knockout of Chrono or Dec1/Dec2 in the Cry1/Cry2 deficient mice did not abolish but decoupled the coherent circadian rhythm into three different periodicities or significantly shortened the circadian period in neonatal SCN. DNA microarray analysis for the SCN of Cry1/Cry2 deficient mice revealed substantial increases in Per(s), Chrono and Dec(s) expression, indicating disinhibition of the transactivation by BMAL1/CLOCK. Here, we conclude that Chrono and Dec1/Dec2 do not compensate for absence of CRY1/CRY2 in the circadian rhythm generation but contribute to the coherent circadian rhythm expression in the neonatal mouse SCN most likely through integration of cellular circadian rhythms.

  Sep. 2021, Scientific reports, 11(1) (1), 19240 - 19240, English, International magazine

  Scientific journal


• Molecular signatures from multi‐omics of autism spectrum disorders and schizophrenia

  Jun Nomura, Matthew Mardo, Toru Takumi

  Wiley, Sep. 2021, Journal of Neurochemistry

  Scientific journal


• Reciprocal differentiation via GABAergic components and ASD-related phenotypes in hES with 1q21.1 CNV

  Yoshiko Nomura, Jun Nomura, Toru Nishikawa, Toru Takumi

  Copy number variations (CNVs) in the distal 1q21.1 region, both deletion (1q del) and duplication (1q dup), are associated with autism spectrum disorder, epilepsy and schizophrenia. Besides common phenotypes, 1q del and 1q dup manifest opposite clinical phenotypes—e.g., microcephaly in 1q del and macrocephaly in 1q dup. However, molecular and cellular mechanisms are still elusive. We generate isogenic human ES (hES) cell lines with reciprocal 1q21.1 CNVs using CRISPR/Cas9 system and differentiate them into 2-dimensional (2-D) neurons and 3-D cortical organoids. Our study recapitulates opposite organoid size and shows dosage-dependent differentiation changes i.e., more mature and GABAergic components in 1q del and more proliferative state in 1q dup. In contrast, both CNVs show hyperexcitability and altered expressions of glutamate system as common features. These results demonstrate that 1q21.1 CNVs dramatically affect cell fate in the early neurodevelopmental periods. This is the first isogenic model of hES CNVs and our findings provide new insights into the underlying mechanisms of neurodevelopmental disorders.

  Cold Spring Harbor Laboratory, Sep. 2021


• Genetic dissection identifies Necdin as a driver gene in a mouse model of paternal 15q duplications.

  Kota Tamada, Keita Fukumoto, Tsuyoshi Toya, Nobuhiro Nakai, Janak R Awasthi, Shinji Tanaka, Shigeo Okabe, François Spitz, Fumihito Saitow, Hidenori Suzuki, Toru Takumi

  Maternally inherited duplication of chromosome 15q11-q13 (Dup15q) is a pathogenic copy number variation (CNV) associated with autism spectrum disorder (ASD). Recently, paternally derived duplication has also been shown to contribute to the development of ASD. The molecular mechanism underlying paternal Dup15q remains unclear. Here, we conduct genetic and overexpression-based screening and identify Necdin (Ndn) as a driver gene for paternal Dup15q resulting in the development of ASD-like phenotypes in mice. An excess amount of Ndn results in enhanced spine formation and density as well as hyperexcitability of cortical pyramidal neurons. We generate 15q dupΔNdn mice with a normalized copy number of Ndn by excising its one copy from Dup15q mice using a CRISPR-Cas9 system. 15q dupΔNdn mice do not show ASD-like phenotypes and show dendritic spine dynamics and cortical excitatory-inhibitory balance similar to wild type animals. Our study provides an insight into the role of Ndn in paternal 15q duplication and a mouse model of paternal Dup15q syndrome.

  Jul. 2021, Nature communications, 12(1) (1), 4056 - 4056, English, International magazine

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Morphological Classification of the Medial Frontal Cortex Based on Cadaver Dissections: A Guide for Interhemispheric Approach.

  Yasutaka Imada, Toru Takumi, Hirohiko Aoyama, Takashi Sadatomo, Kaoru Kurisu

  The medial frontal cortex (MFC) is a part of the medial surface of the frontal lobe situated in the rostral portion of the corpus callosum (CC). In a surgical interhemispheric approach (IHA), the MFC covers the anterior communicating artery (Aco) complex until the final stage of dissection. To clarify the anatomical relationship between the MFC and the Aco complex, and to facilitate orientation in IHA, we analyzed the morphological features of the MFC in number, size, and pattern of gyri from the medial surface of the hemisphere in the subcallosal portion using 53 adult cadaveric hemispheres. The mean width of the MFC excluding cingulate gyrus (MFCexcg) was 20.6 ± as mm in the subcallosal portion. MFCexcg consisting of 2, 3, 4, or 5 gyri were observed in 7.5%, 56.6%, 32.1%, or 3.8% of the hemispheres, respectively. Bilateral MFCexcg consisting of >2 gyri were observed in approximately 85% of the hemispheres. Therefore, in many cases, the dissection performed at 2 cm upward from the base of the straight gyrus (SG) or 3-4 gyri of the MFC is sufficient to safely reach the upper portion of the cistern of lamina terminalis located distal to the Aco complex in IHA. The MFC is a good landmark for intraoperative orientation in IHA.

  May 2021, Neurologia medico-chirurgica, 61(5) (5), 302 - 311, English, Domestic magazine

  Scientific journal


• Transcriptome analysis of human neural cells derived from isogenic embryonic stem cells with 16p11.2 deletion

  Yoshiko Nomura, Jun Nomura, Hiroyuki Kamiguchi, Toru Nishikawa, Toru Takumi

  Elsevier BV, Mar. 2021, Neuroscience Research

  Scientific journal


• Cranioplastic Surgery and Acclimation Training for Awake Mouse fMRI

  Tomokazu Tsurugizawa, Kota Tamada, Clement Debacker, Andrew Zalesky, Toru Takumi

  Bio-Protocol, LLC, 2021, BIO-PROTOCOL, 11(7) (7)

  Scientific journal


• Optogenetic Approaches to Understand the Neural Circuit Mechanism of Social Deficits Seen in Autism Spectrum Disorders.

  Nobuhiro Nakai, Eric T N Overton, Toru Takumi

  Individuals with neurodevelopmental disorders, such as autism spectrum disorders (ASDs), are diagnosed based on nonquantitative objective parameters such as behavioral phenotypes. It is still unclear how any neural mechanism affects such behavioral phenotypes in these patients. In human genetics, a large number of genetic abnormalities including single nucleotide variation (SNV) and copy number variation (CNV) have been found in individuals with ASDs. It is thought that influence of such variations converges on dysfunction of neural circuit resulting in common behavioral phenotypes of ASDs such as deficits in social communication and interaction. Recent studies suggest that an excitatory/inhibitory (E/I) imbalanced state, which induces disruption of neural circuit activities, is one of the pathophysiological abnormalities in ASD brains. To assess the causal relationship between brain abnormalities and behavioral deficits, we can take advantage of optogenetics with animal models of ASDs that recapitulate human genetic mutations. Here, we review optogenetics studies being utilized to dissect neural circuit mechanisms associated with social deficits in model mice of ASD. Optogenetic manipulation of disrupted neural activities would help us understand how neural circuits affect behavioral deficits observed in ASDs.

  2021, Advances in experimental medicine and biology, 1293, 523 - 533, English, International magazine

  Scientific journal


• Development of serotonergic projections to the suprachiasmatic nucleus in the mouse brain.

  Janak R Awasthi, Kota Tamada, Eric T N Overton, Toru Takumi

  Serotonin (5-HT) and its innervation have been implicated in various neural functions including circadian systems. Although classical studies have examined the 5-HT innervation pattern in the adult suprachiasmatic nucleus (SCN), the fine-grained morphological study of the development of pathway and terminal projections to the SCN remains scarce. Here, we utilize transgenic mice expressing GFP under the serotonin transporter (SERT) promoter to subserve our developmental mapping study. We demonstrate that the morphology of 5-HT pathway fibers decussating over the supraoptic commissure that projects to the SCN exhibits two distinct developmental patterns. The punctate fibers at the fetal stage gradually become smooth and filamentous, especially during postnatal one week and remain constant thereafter. The innervation field in the SCN develops properly only during postnatal two weeks. Its ventromedial area remains one of the highest 5-HT innervated areas in the adult brain, whereas the dorsolateral area is less innervated. Thus, we provide novel and specific insights on the developmental map of 5-HT system into the SCN using transgenic mouse.

  Nov. 2020, Neuroscience letters, 739, 135438 - 135438, English, International magazine

  Scientific journal


• Imaging the Neural Circuit Basis of Social Behavior: Insights from Mouse and Human Studies.

  Isamu Miura, Eric T N Overton, Nobuhiro Nakai, Takakazu Kawamata, Masaaki Sato, Toru Takumi

  Social behavior includes a variety of behaviors that are expressed between two or more individuals. In humans, impairment of social function (i.e., social behavior and social cognition) is seen in neurodevelopmental and neurological disorders including autism spectrum disorders (ASDs) and stroke, respectively. In basic neuroscience research, fluorescence monitoring of neural activity, such as immediate early gene (IEG)-mediated whole-brain mapping, fiber photometry, and calcium imaging using a miniaturized head-mounted microscope or a two-photon microscope, and non-fluorescence imaging such as functional magnetic resonance imaging (fMRI) are increasingly used to measure the activity of many neurons and multiple brain areas in animals during social behavior. In this review, we overview recent rodent studies that have investigated the dynamics of brain activity during social behavior at the whole-brain and local circuit levels and studies that explored the neural basis of social function in healthy, in brain-injured, and in autistic human subjects. A synthesis of such findings will advance our understanding of brain mechanisms underlying social behavior and facilitate the development of pharmaceutical and functional neurosurgical interventions for brain disorders affecting social function.

  Sep. 2020, Neurologia medico-chirurgica, 60(9) (9), 429 - 438, English, Domestic magazine

  [Refereed]

  Scientific journal


• Comprehensive topographical map of the serotonergic fibers in the male mouse brain.

  Janak R Awasthi, Kota Tamada, Eric T N Overton, Toru Takumi

  It is well established that serotonergic fibers distribute throughout the brain. Abnormal densities or patterns of serotonergic fibers have been implicated in neuropsychiatric disorders. Although many classical studies have examined the distribution pattern of serotonergic fibers, most of them were either limited to specific brain areas or had limitations in demonstrating the fine axonal morphology. In this study, we utilize male mice expressing green fluorescence protein under the serotonin transporter (SERT) promoter to map the topography of serotonergic fibers across the rostro-caudal extent of each brain area. We demonstrate previously unreported regional density and fine-grained anatomy of serotonergic fibers. Our findings include: (a) SERT fibers distribute abundantly in the thalamic nuclei close to the midline and dorsolateral areas, in most of the hypothalamic nuclei with few exceptions such as the median eminence and arcuate nuclei, and within the basal amygdaloid complex and lateral septal nuclei, (b) the source fibers of innervation of the hippocampus traverse through the septal nuclei before reaching its destination, (c) unique, filamentous type of straight terminal fibers within the nucleus accumbens, (d) laminar pattern of innervation in the hippocampus, olfactory bulb and cortex with heterogenicity in innervation density among the layers, (e) cortical labeling density gradually decreases rostro-caudally, (f) fibers traverse and distribute mostly within the gray matter, leaving the white fiber bundles uninnervated, and (g) most of the highly labeled nuclei and cortical areas have predominant anatomical connection to limbic structures. In conclusion, we provide novel, regionally specific insights on the distribution map of serotonergic fibers using transgenic mouse.

  Wiley, Sep. 2020, The Journal of comparative neurology, 529(7) (7), 1391 - 1429, English, International magazine

  Scientific journal


• Encoding of social exploration by neural ensembles in the insular cortex.

  Isamu Miura, Masaaki Sato, Eric T N Overton, Nobuo Kunori, Junichi Nakai, Takakazu Kawamata, Nobuhiro Nakai, Toru Takumi

  The insular cortex (IC) participates in diverse complex brain functions, including social function, yet their cellular bases remain to be fully understood. Using microendoscopic calcium imaging of the agranular insular cortex (AI) in mice interacting with freely moving and restrained social targets, we identified 2 subsets of AI neurons-a larger fraction of "Social-ON" cells and a smaller fraction of "Social-OFF" cells-that change their activity in opposite directions during social exploration. Social-ON cells included those that represented social investigation independent of location and consisted of multiple subsets, each of which was preferentially active during exploration under a particular behavioral state or with a particular target of physical contact. These results uncover a previously unknown function of AI neurons that may act to monitor the ongoing status of social exploration while an animal interacts with unfamiliar conspecifics.

  Sep. 2020, PLoS biology, 18(9) (9), e3000584, English, International magazine

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Upregulated 5-HT1A receptor-mediated currents in the prefrontal cortex layer 5 neurons in the 15q11-13 duplication mouse model of autism.

  Fumihito Saitow, Toru Takumi, Hidenori Suzuki

  Serotonin (5-HT) is a well-known modulator of behavioral, physiological, and emotional functions of the forebrain region. We recently discovered alterations of serotonergic synaptic modulations in both, the prefrontal cortex (PFC) and the somatosensory cortex, in the 15q dup mouse model of autism spectrum disorder (ASD). To further understand the roles of the 5-HT system implicated in developmental disorders such as ASD, comparison with model animals exhibiting different phenotypes may be useful. In this study, we investigated the relationship between sociability and the magnitude of 5-HT1A receptor (5-HT1AR) activation-induced outward currents from layer 5 pyramidal neurons in the PFC, because a mouse model of Williams-Beuren syndrome (WBS; another developmental disorder exhibiting low innate anxiety and high sociability) reportedly showed larger 5-HT-induced currents. To investigate whether the 5-HT1AR activation-induced outward currents are involved in the endophenotype determination of social behavior, we examined 15q dup mice with a phenotype opposite to WBS. We found 5-HT elicited significantly larger outward currents in 15q dup mice than in WT controls, regardless of sociability. In contrast, baclofen-induced GABAB receptor-mediated outward currents were not significantly different between genotypes, although GABAB receptor was coupled to Gi/o as well as 5-HT1A. Further, we found the larger 5-HT1AR-mediated currents in 15q dup mice did not affect the magnitude of inhibitory action of NMDA receptor functions. Taken together, our results provide a potential physiological hallmark for developmental disorders that may involve the imbalance of the neuronal circuity in the PFC.

  Springer Science and Business Media LLC, Aug. 2020, Molecular brain, 13(1) (1), 115 - 115, English, International magazine

  Scientific journal


• Imaging the neural circuit basis of social behavior: insights from mouse and human studies

  Miura I, Overton ETN, Nakai N, Kawamata T, Sato M, Takumi T

  Jun. 2020, Neurol. Med. Chir (Tokyo), in press

  [Refereed]


• Behavioral neuroscience of autism.

  Takumi T, Tamada K, Hatanaka F, Nakai N, Bolton PF

  Autism spectrum disorder (ASD) is a neurodevelopmental disorder. Several genetic causes of ASD have been identified and this has enabled researchers to construct mouse models. Mouse behavioral tests reveal impaired social interaction and communication, as well as increased repetitive behavior and behavioral inflexibility in these mice, which correspond to core behavioral deficits observed in individuals with ASD. However, the connection between these behavioral abnormalities and the underlying dysregulation in neuronal circuits and synaptic function is poorly understood. Moreover, different components of the ASD phenotype may be linked to dysfunction in different brain regions, making it even more challenging to chart the pathophysiological mechanisms involved in ASD. Here we summarize the research on mouse models of ASD and their contribution to understanding pathophysiological mechanisms. Specifically, we emphasize abnormal serotonin production and regulation, as well as the disruption in circadian rhythms and sleep that are observed in a subset of ASD, and propose that spatiotemporal disturbances in brainstem development may be a primary cause of ASD that propagates towards the cerebral cortex.

  Mar. 2020, Neuroscience and biobehavioral reviews, 110, 60 - 76, English, International magazine, Co-authored internationally

  [Refereed]

  Scientific journal


• Awake functional MRI detects neural circuit dysfunction in a mouse model of autism

  Tomokazu Tsurugizawa, Kota Tamada, Nobukazu Ono, Sachise Karakawa, Yuko Kodama, Clement Debacker, Junichi Hata, Hideyuki Okano, Akihiko Kitamura, Andrew Zalesky, Toru Takumi

  MRI has potential as a translational approach from rodents to humans. However, given that mouse functional MRI (fMRI) uses anesthetics for suppression of motion, it has been difficult to directly compare the result of fMRI in “unconsciousness” disease model mice with that in “consciousness” patients. We develop awake fMRI to investigate brain function in 15q dup mice, a copy number variation model of autism. Compared to wild-type mice, we find that 15q dup is associated with whole-brain functional hypoconnectivity and diminished fMRI responses to odors of stranger mice. Ex vivo diffusion MRI reveals widespread anomalies in white matter ultrastructure in 15q dup mice, suggesting a putative anatomical substrate for these functional hypoconnectivity. We show that d-cycloserine (DCS) treatment partially normalizes these anormalies in the frontal cortex of 15q dup mice and rescues some social behaviors. Our results demonstrate the utility of awake rodent fMRI and provide a rationale for further investigation of DCS therapy.

  American Association for the Advancement of Science (AAAS), Feb. 2020, Science Advances, 6(6) (6), eaav4520 - eaav4520, English, International magazine, Co-authored internationally

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Change in serotonergic modulation contributes to the synaptic imbalance of neuronal circuit at the prefrontal cortex in the 15q11-13 duplication mouse model of autism

  Saitow, F., Takumi, T., Suzuki, H.

  2020, Neuropharmacology, 165, English

  [Refereed]

  Scientific journal


• Altered microbiota composition reflects enhanced communication in 15q11-13 CNV mice

  Septyaningtrias DE, Lin CW, Ouchida R, Nakai N, Suda W, Hattori M, Morita H, Honda K, Tamada K, Takumi T

  Autism spectrum disorder (ASD) is a complex and heterogeneous neurodevelopmental disorder. In addition to the core symptoms of ASD, many patients with ASD also show comorbid gut dysbiosis, which may lead to various gastrointestinal (GI) problems. Intriguingly, there is evidence that gut microbiota communicate with the central nervous system to modulate behavioral output through the gut-brain axis. To investigate how the microbiota composition is changed in ASD and to identify which microbes are involved in autistic behaviors, we performed a 16S rRNA gene-based metagenomics analysis in an ASD mouse model. Here, we focused on a model with human 15q11-13 duplication (15q dup), the most frequent chromosomal aberration or copy number variation found in ASD. Species diversity of the microbiome was significantly decreased in 15q dup mice. A combination of antibiotics treatment and behavioral analysis showed that neomycin improved social communication in 15q dup mice. Furthermore, comparison of the microbiota composition of mice treated with different antibiotics enabled us to identify beneficial operational taxonomic units (OTUs) for ultrasonic vocalization.

  2020, Neurosci Res, 161, 59 - 67, English, International magazine

  [Refereed]

  Scientific journal


• Behavioral analysis in mice deficient for GAREM2 (Grb2-associated regulator of Erk/MAPK subtype2) that is a subtype of highly expressing in the brain.

  Nishino T, Tamada K, Maeda A, Abe T, Kiyonari H, Funahashi Y, Kaibuchi K, Takumi T, Konishi H

  Grb2-associated regulator of Erk/MAPK (GAREM), is an adaptor protein related to the several cell growth factor receptor-signaling. The GAREM family has two subtypes, GAREM1 and GAREM2, both encoded in the human and mouse genome. Recent genome-wide research identified GAREM2 as a candidate of neurodegenerative diseases. Here, we use knockout (KO) mice to show the role of GAREM2, that is highly expressed in the brain. According to the comprehensive behavioral battery, they exhibited less anxiety both in elevated plus maze and open field tests, mildly increased social approaching behavior in the reciprocal social interaction test, and longer latency to immobility in the tail suspension test as compared to wild-type (WT). Additionally, the extension of neurites in the primary cultured neurons was suppressed in ones derived from GAREM2 KO mice. Furthermore, we also identified Intersectin, as a binding partner of GAREM2 in this study. Intersectin is also a multi-domain adaptor protein that regulates endocytosis and cell signaling, which can potentially alter the subcellular localization of GAREM2. The important molecules, such as the neurotrophin receptor and Erk family, that are involved in the signaling pathway of the neural cell growth in the mouse brain, have been reported to participate in emotional behavior. As GAREM plays a role in the cellular growth factor receptor signaling pathway, GAREM2 may have a common role related to the transduction of Erk signaling in the higher brain functions.

  Nov. 2019, Molecular brain, 12(1) (1), 94 - 94, English, International magazine

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Recent genetic and functional insights in autism spectrum disorder

  Nakanishi, M., Anderson, M.P., Takumi, T.

  Aug. 2019, Current Opinion in Neurology, 32(4) (4), 627 - 634, Co-authored internationally

  [Refereed]

  Scientific journal


• Morphological pattern and classification of the superficial middle cerebral vein by cadaver dissections: An embryological viewpoint

  Imada, Y., Kurisu, K., Takumi, T., Aoyama, H., Sadatomo, T., Migita, K., Yuki, K.

  Jul. 2019, Neurologia Medico Chirurgica, 59(7) (7), 264 - 270

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• UBE3A regulates the transcription of IRF, an antiviral immunity.

  Furumai R, Tamada K, Liu X, Takumi T

  UBE3A is a gene responsible for the pathogenesis of Angelman syndrome (AS), a neurodevelopmental disorder characterized by symptoms such as intellectual disability, delayed development and severe speech impairment. UBE3A encodes an E3 ubiquitin ligase, for which several targets have been identified, including synaptic molecules. Although proteolysis mainly occurs in the cytoplasm, UBE3A is localized to the cytoplasm and the nucleus. In fact, UBE3A is also known as a transcriptional regulator of the family of nuclear receptors. However, the function of UBE3A in the nucleus remains unclear. Therefore, we examined the involvement of UBE3A in transcription in the nuclei of neurons. Genome-wide transcriptome analysis revealed an enrichment of genes downstream of interferon regulatory factor (IRF) in a UBE3A-deficient AS mouse model. In vitro biochemical analyses further demonstrated that UBE3A interacted with IRF and, more importantly, that UBE3A enhanced IRF-dependent transcription. These results suggest a function for UBE3A as a transcriptional regulator of the immune system in the brain. These findings also provide informative molecular insights into the function of UBE3A in the brain and in AS pathogenesis.

  Jun. 2019, Human molecular genetics, 28(12) (12), 1947 - 1958, English, International magazine

  [Refereed]

  Scientific journal


• Gene expression profile data of the developing small intestine of Id2-deficient mice.

  Kentaro Mori, Kota Tamada, Hisanori Kurooka, Makoto Matsui, Toru Takumi, Yoshifumi Yokota

  This article contains data related to the research article entitled "Id2 determines intestinal identity through repression of the foregut transcription factor, Irx5" [1]. Id2 deficient (Id2-/-) mice developed gastric tumors and heterotopic squamous epithelium in the small intestine. These tumors and heterotopic tissues were derived from ectopic gastric cells and squamous cells formed in the small intestine respectively during development. In this study, microarray data of the developing small intestine of Id2-/- mice was analyzed.

  Jun. 2019, Data in brief, 24, 103717 - 103717, English, International magazine

  [Refereed]

  Scientific journal


• UBE3A-mediated PTPA ubiquitination and degradation regulate PP2A activity and dendritic spine morphology

  Wang, J., Lou, S.-S., Wang, T., Wu, R.-J., Li, G., Zhao, M., Lu, B., Li, Y.-Y., Zhang, J., Cheng, X., Shen, Y., Wang, X., Zhu, Z.-C., Li, M.-J., Takumi, T., Yang, H., Yu, X., Liao, L., Xiong, Z.-Q.

  Jun. 2019, Proceedings of the National Academy of Sciences of the United States of America, 116(25) (25), 12500 - 12505, Co-authored internationally

  [Refereed]

  Scientific journal


• Fetal neural stem cells from a mouse model of 15q11-13 duplication syndrome exhibit altered differentiation into neurons and astrocytes

  Choi, Y., Kim, H., Choi, M., Yang, E.-J., Takumi, T., Kim, H.-S.

  Mar. 2019, Journal of Pharmacological Sciences, 139(3) (3), 249 - 253, Co-authored internationally

  [Refereed]

  Scientific journal


• Author Correction: The choroid plexus is an important circadian clock component (Nature Communications, (2018), 9, 1, (1062), 10.1038/s41467-018-03507-2)

  Myung, J., Schmal, C., Hong, S., Tsukizawa, Y., Rose, P., Zhang, Y., Holtzman, M.J., De Schutter, E., Herzel, H., Bordyugov, G., Takumi, T.

  2019, Nature Communications, 10(1) (1)

  Scientific journal


• The choroid plexus is an important circadian clock component

  Jihwan Myung, Christoph Schmal, Sungho Hong, Yoshiaki Tsukizawa, Pia Rose, Yong Zhang, Michael J. Holtzman, Erik De Schutter, Hanspeter Herzel, Grigory Bordyugov, Toru Takumi

  Nature Publishing Group, Dec. 2018, Nature Communications, 9(1) (1), 1062, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Glutamate and GABA in autism spectrum disorder-a translational magnetic resonance spectroscopy study in man and rodent models

  Jamie Horder, Marija M. Petrinovic, Maria A. Mendez, Andreas Bruns, Toru Takumi, Will Spooren, Gareth J. Barker, Basil Künnecke, Declan G. Murphy

  Nature Publishing Group, Dec. 2018, Translational Psychiatry, 8(1) (1), 106, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Quantitative evaluation of incomplete preweaning lethality in mice by using the CRISPR/Cas9 system

  Takumi Nakamura, Kazuo Nakajima, Tetsuo Ohnishi, Takeo Yoshikawa, Moe Nakanishi, Toru Takumi, Takashi Tsuboi, Tadafumi Kato

  Dec. 2018, Scientific Reports, 8(1) (1), 16025

  [Refereed]

  Scientific journal


• Critical roles of serotonin-oxytocin interaction during the neonatal period in social behavior in 15q dup mice with autistic traits

  Nagano, M., Takumi, T., Suzuki, H.

  Sep. 2018, Scientific Reports, 8(1) (1), 13675

  [Refereed]

  Scientific journal


• Common defects of spine dynamics and circuit function in neurodevelopmental disorders: A systematic review of findings from in vivo optical imaging of mouse models

  Nobuhiro Nakai, Toru Takumi, Junichi Nakai, Masaaki Sato

  Frontiers Media S.A., Jun. 2018, Frontiers in Neuroscience, 12, 412, English

  [Refereed]


• Postsynaptic density proteins and their involvement in neurodevelopmental disorders

  Takeshi Kaizuka, Toru Takumi

  Oxford University Press, Jun. 2018, Journal of Biochemistry, 163(6) (6), 447 - 455, English

  [Refereed]

  Scientific journal


• Id2 Determines Intestinal Identity through Repression of the Foregut Transcription Factor Irx5.

  Kentaro Mori, Harumi Nakamura, Hisanori Kurooka, Hitoshi Miyachi, Kota Tamada, Manabu Sugai, Toru Takumi, Yoshifumi Yokota

  The cellular components and function of the gastrointestinal epithelium exhibit distinct characteristics depending on the region, e.g., stomach or intestine. How these region-specific epithelial characteristics are generated during development remains poorly understood. Here, we report on the involvement of the helix-loop-helix inhibitor Id2 in establishing the specific characteristics of the intestinal epithelium. Id2-/- mice developed tumors in the small intestine. Histological analysis indicated that the intestinal tumors were derived from gastric metaplasia formed in the small intestine during development. Heterotopic Id2 expression in developing gastric epithelium induced a fate change to intestinal epithelium. Gene expression analysis revealed that foregut-enriched genes encoding Irx3 and Irx5 were highly induced in the midgut of Id2-/- embryos, and transgenic mice expressing Irx5 in the midgut endoderm developed tumors recapitulating the characteristics of Id2-/- mice. Altogether, our results demonstrate that Id2 plays a crucial role in the development of regional specificity in the gastrointestinal epithelium.

  May 2018, Molecular and cellular biology, 38(9) (9), English, International magazine

  [Refereed]

  Scientific journal


• A novel role of the antitumor agent tricyclodecan-9-yl-xanthogenate as an open channel blocker of KCNQ1/KCNE1

  Meikui Wu, Makoto Takemoto, Huan Luo, Jian-Jun Xu, Mei-Hong Lu, Masaki Kameyama, Toru Takumi, Wen-Jie Song

  Elsevier B.V., Apr. 2018, European Journal of Pharmacology, 824, 99 - 107, English

  [Refereed]

  Scientific journal


• CNV biology in neurodevelopmental disorders

  Toru Takumi, Kota Tamada

  Elsevier Ltd, Feb. 2018, Current Opinion in Neurobiology, 48, 183 - 192, English

  [Refereed]


• Identification of genes regulating GABAergic interneuron maturation.

  Fukumoto K, Tamada K, Toya T, Nishino T, Yanagawa Y, Takumi T

  During embryonic development, GABAergic interneurons, a main inhibitory component in the cerebral cortex, migrate tangentially from the ganglionic eminence (GE) to cerebral cortex. After reaching the cerebral cortex, they start to extend their neurites for constructing local neuronal circuits around the neonatal stage. Aberrations in migration or neurite outgrowth are implicated in neurological and psychiatric disorders such as epilepsy, schizophrenia and autism. Previous studies revealed that in the early phase of cortical development the neural population migrates tangentially from the GE in the telencephalon and several genes have been characterized as regulators of migration and specification of GABAergic interneurons. However, much less is known about the molecular mechanisms of GABAergic interneurons-specific maturation at later stages of development. Here, we performed genome-wide screening to identify genes related to the later stage by flow cytometry based-microarray (FACS-array) and identified 247 genes expressed in cortical GABAergic interneurons. Among them, Dgkg, a member of diacylglycerol kinase family, was further analyzed. Correlational analysis revealed that Dgkg is dominantly expressed in somatostatin (SST)-expressing GABAergic interneurons. The functional study of Dgkg using GE neurons indicated alteration in neurite outgrowth of GABAergic neurons. This study shows a new functional role for Dgkg in GABAergic interneurons as well as the identification of other candidate genes for their maturation.

  Dec. 2017, Neuroscience research, 134, 18 - 29, English, International magazine

  [Refereed]

  Scientific journal


• Rodent models of genetic and chromosomal variations in psychiatric disorders

  Jun Nomura, Geetha Kannan, Toru Takumi

  Aug. 2017, PSYCHIATRY AND CLINICAL NEUROSCIENCES, 71(8) (8), 508 - 517, English, Co-authored internationally

  [Refereed]


• Potential contribution of tandem circadian enhancers to nonlinear oscillations in clock gene expression

  Isao T. Tokuda, Akihiko Okamoto, Ritsuko Matsumura, Toru Takumi, Makoto Akashi

  Aug. 2017, MOLECULAR BIOLOGY OF THE CELL, 28(17) (17), 2333 - 2342, English

  [Refereed]

  Scientific journal


• Functional significance of rare neuroligin 1 variants found in autism

  Moe Nakanishi, Jun Nomura, Xiao Ji, Kota Tamada, Takashi Arai, Eiki Takahashi, Maja Bućan, Toru Takumi

  Public Library of Science, Aug. 2017, PLoS Genetics, 13(8) (8), e1006940, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Skeletal Site-specific Changes in Bone Mass in a Genetic Mouse Model for Human 15q11-13 Duplication Seen in Autism

  Kirsty E. Lewis, Kunal Sharan, Toru Takumi, Vijay K. Yadav

  Aug. 2017, SCIENTIFIC REPORTS, 7(1) (1), 9902, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Serotonin rebalances cortical tuning and behavior linked to autism symptoms in 15q11-13 CNV mice

  Nobuhiro Nakai, Masatoshi Nagano, Fumihito Saitow, Yasuhito Watanabe, Yoshinobu Kawamura, Akiko Kawamoto, Kota Tamada, Hiroshi Mizuma, Hirotaka Onoe, Yasuyoshi Watanabe, Hiromu Monai, Hajime Hirase, Jin Nakatani, Hirofumi Inagaki, Tomoyuki Kawada, Taisuke Miyazaki, Masahiko Watanabe, Yuka Sato, Shigeo Okabe, Kazuo Kitamura, Masanobu Kano, Kouichi Hashimoto, Hidenori Suzuki, Toru Takumi

  Jun. 2017, SCIENCE ADVANCES, 3(6) (6), e1603001, English

  [Refereed]

  Scientific journal


• Behavioral and neuroanatomical analyses in a genetic mouse model of 2q13 duplication

  Keiko Kishimoto, Jun Nomura, Jacob Ellegood, Keita Fukumoto, Jason P. Lerch, Daniel Moreno-De-Luca, Thomas Bourgeron, Kota Tamada, Toru Takumi

  May 2017, GENES TO CELLS, 22(5) (5), 436 - 451, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Unusual semi-extractability as a hallmark of nuclear body-associated architectural noncoding RNAs

  Takeshi Chujo, Tomohiro Yamazaki, Tetsuya Kawaguchi, Satoshi Kurosaka, Toru Takumi, Shinichi Nakagawa, Tetsuro Hirose

  May 2017, EMBO JOURNAL, 36(10) (10), 1447 - 1462, English

  [Refereed]

  Scientific journal


• Distinct Defects in Spine Formation or Pruning in Two Gene Duplication Mouse Models of Autism

  Miao Wang, Huiping Li, Toru Takumi, Zilong Qiu, Xiu Xu, Xiang Yu, Wen-Jie Bian

  Apr. 2017, NEUROSCIENCE BULLETIN, 33(2) (2), 143 - 152, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Ror2 signaling regulates Golgi structure and transport through IFT20 for tumor invasiveness.

  Michiru Nishita, Seung-Yeol Park, Tadashi Nishio, Koki Kamizaki, ZhiChao Wang, Kota Tamada, Toru Takumi, Ryuju Hashimoto, Hiroki Otani, Gregory J Pazour, Victor W Hsu, Yasuhiro Minami

  Signaling through the Ror2 receptor tyrosine kinase promotes invadopodia formation for tumor invasion. Here, we identify intraflagellar transport 20 (IFT20) as a new target of this signaling in tumors that lack primary cilia, and find that IFT20 mediates the ability of Ror2 signaling to induce the invasiveness of these tumors. We also find that IFT20 regulates the nucleation of Golgi-derived microtubules by affecting the GM130-AKAP450 complex, which promotes Golgi ribbon formation in achieving polarized secretion for cell migration and invasion. Furthermore, IFT20 promotes the efficiency of transport through the Golgi complex. These findings shed new insights into how Ror2 signaling promotes tumor invasiveness, and also advance the understanding of how Golgi structure and transport can be regulated.

  Jan. 2017, Scientific reports, 7(1) (1), 1 - 1, English, International magazine, Co-authored internationally

  [Refereed]

  Scientific journal


• CHRONO integrates behavioral stress and epigenetic control of metabolism

  Fumiyuki Hatanaka, Toru Takumi

  2017, ANNALS OF MEDICINE, 49(4) (4), 352 - 356, English

  [Refereed]


• Network Dynamics Mediate Circadian Clock Plasticity

  Abdelhalim Azzi, Jennifer A. Evans, Tanya Leise, Jihwan Myung, Toru Takumi, Alec J. Davidson, Steven A. Brown

  Jan. 2017, NEURON, 93(2) (2), 441 - 450, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Exome sequencing in the knockin mice generated using the CRISPR/Cas system

  Kazuo Nakajima, An-a Kazuno, John Kelsoe, Moe Nakanishi, Toru Takumi, Tadafumi Kato

  Oct. 2016, SCIENTIFIC REPORTS, 6, 34703, English

  [Refereed]

  Scientific journal


• FUS/TLS acts as an aggregation-dependent modifier of polyglutamine disease model mice

  Yoshihiro Kino, Chika Washizu, Masaru Kurosawa, Mizuki Yamada, Hiroshi Doi, Toru Takumi, Hiroaki Adachi, Masahisa Katsuno, Gen Sobue, Geoffrey G. Hicks, Nobutaka Hattori, Tomomi Shimogori, Nobuyuki Nukina

  Oct. 2016, SCIENTIFIC REPORTS, 6, 35236, English, Co-authored internationally

  [Refereed]

  Scientific journal


• CHD8 haploinsufficiency results in autistic-like phenotypes in mice

  Yuta Katayama, Masaaki Nishiyama, Hirotaka Shoji, Yasuyuki Ohkawa, Atsuki Kawamura, Tetsuya Sato, Mikita Suyama, Toru Takumi, Tsuyoshi Miyakawa, Keiichi I. Nakayama

  Sep. 2016, NATURE, 537(7622) (7622), 675 - +, English

  [Refereed]

  Scientific journal


• Structural, super-resolution microscopy analysis of paraspeckle nuclear body organization

  Jason A. West, Mari Mito, Satoshi Kurosaka, Toru Takumi, Chiharu Tanegashima, Takeshi Chujo, Kaori Yanaka, Robert E. Kingston, Tetsuro Hirose, Charles Bond, Archa Fox, Shinichi Nakagawa

  Sep. 2016, JOURNAL OF CELL BIOLOGY, 214(7) (7), 817 - 830, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Regulation of membrane KCNQ1/KCNE1 channel density by sphingomyelin synthase 1

  Meikui Wu, Makoto Takemoto, Makoto Taniguchi, Toru Takumi, Toshiro Okazaki, Wen-Jie Song

  Jul. 2016, AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY, 311(1) (1), C15 - C23, English

  [Refereed]

  Scientific journal


• Translocated in liposarcoma regulates the distribution and function of mammalian enabled, a modulator of actin dynamics

  Tomohito Sugiura, Shuji Matsuda, Satoshi Kurosaka, Nobuhiro Nakai, Keita Fukumoto, Tetsuya Takahashi, Hirofumi Maruyama, Kazunori Imaizumi, Masayasu Matsumoto, Toru Takumi

  Apr. 2016, FEBS JOURNAL, 283(8) (8), 1475 - 1487, English

  [Refereed]

  Scientific journal


• Role for neonatal D-serine signaling: prevention of physiological and behavioral deficits in adult Pickl knockout mice

  J. Nomura, H. Jaaro-Peled, E. Lewis, P. Nunez-Abades, F. Huppe-Gourgues, T. Cash-Padgett, F. Emiliani, M. A. Kondo, A. Furuya, M. A. Landek-Salgado, Y. Ayhan, A. Kamiya, T. Takumi, R. Huganir, M. Pletnikov, P. O'Donnell, A. Sawa

  Mar. 2016, MOLECULAR PSYCHIATRY, 21(3) (3), 386 - 393, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Sequence features associated with the cleavage efficiency of CRISPR/Cas9 system

  Xiaoxi Liu, Ayaka Homma, Jamasb Sayadi, Shu Yang, Jun Ohashi, Toru Takumi

  Jan. 2016, SCIENTIFIC REPORTS, 6, 19675, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Neuroanatomical Phenotypes Are Consistent With Autism-Like Behavioral Phenotypes in the 15q11-13 Duplication Mouse Model

  Jacob Ellegood, Nobuhiro Nakai, Jin Nakatani, Mark Henkelman, Toru Takumi, Jason Lerch

  Oct. 2015, AUTISM RESEARCH, 8(5) (5), 545 - 555, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Autism spectrum disorder model mice: Focus on copy number variation and epigenetics

  Nobuhiro Nakai, Susumu Otsuka, Jihwan Myung, Toru Takumi

  Oct. 2015, SCIENCE CHINA-LIFE SCIENCES, 58(10) (10), 976 - 984, English

  [Refereed]


• Constant light enhances synchrony among circadian clock cells and promotes behavioral rhythms in VPAC(2)-signaling deficient mice

  Alun T. L. Hughes, Cara. L. Croft, Rayna E. Samuels, Jihwan Myung, Toru Takumi, Hugh D. Piggins

  Sep. 2015, SCIENTIFIC REPORTS, 5, 14044, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Transcriptome profiling of white adipose tissue in a mouse model for 15q duplication syndrome

  Xiaoxi Liu, Kota Tamada, Rui Kishimoto, Hiroko Okubo, Satoko Ise, Hisashi Ohta, Sandra Ruf, Jin Nakatani, Nobuoki Kohno, François Spitz, Toru Takumi

  Elsevier Inc, Sep. 2015, Genomics Data, 5, 394 - 396, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Model mice for 15q11-13 duplication syndrome exhibit late-onset obesity and altered lipid metabolism

  Rui Kishimoto, Kota Tamada, Xiaoxi Liu, Hiroko Okubo, Satoko Ise, Hisashi Ohta, Sandra Ruf, Jin Nakatani, Nobuoki Kohno, Francois Spitz, Toru Takumi

  Aug. 2015, HUMAN MOLECULAR GENETICS, 24(16) (16), 4559 - 4572, English, Co-authored internationally

  [Refereed]

  Scientific journal


• GABA-mediated repulsive coupling between circadian clock neurons in the SCN encodes seasonal time

  Jihwan Myung, Sungho Hong, Daniel DeWoskin, Erik De Schutter, Daniel B. Forger, Toru Takumi

  Jul. 2015, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 112(29) (29), E3920 - E3929, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Distinct roles for GABA across multiple timescales in mammalian circadian timekeeping

  Daniel DeWoskin, Jihwan Myung, Mino D. C. Belle, Hugh D. Piggins, Toru Takumi, Daniel B. Forger

  Jul. 2015, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 112(29) (29), E3911 - E3919, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Cerebellar associative sensory learning defects in five mouse autism models

  Alexander D. Kloth, Aleksandra Badura, Amy Li, Adriana Cherskov, Sara G. Connolly, Andrea Giovannucci, M. Ali Bangash, Giorgio Grasselli, Olga Penagarikano, Claire Piochon, Peter T. Tsai, Daniel H. Geschwind, Christian Hansel, Mustafa Sahin, Toru Takumi, Paul F. Worley, Samuel S-H Wang

  Jul. 2015, ELIFE, 4, e06085, English, Co-authored internationally

  [Refereed]

  Scientific journal


• DEC2-E4BP4 heterodimer represses the transcriptional enhancer activity of the EE element in the Per2 promoter

  Shintaro Tanoue, Katsumi Fujimoto, Jihwan Myung, Fumiyuki Hatanaka, Yukio Kato, Toru Takumi

  Jul. 2015, FRONTIERS IN NEUROLOGY, 6, 166, English

  [Refereed]

  Scientific journal


• FUS/TLS deficiency causes behavioral and pathological abnormalities distinct from amyotrophic lateral sclerosis

  Yoshihiro Kino, Chika Washizu, Masaru Kurosawa, Mizuki Yamada, Haruko Miyazaki, Takumi Akagi, Tsutomu Hashikawa, Hiroshi Doi, Toru Takumi, Geoffrey G. Hicks, Nobutaka Hattori, Tomomi Shimogori, Nobuyuki Nukina

  Apr. 2015, ACTA NEUROPATHOLOGICA COMMUNICATIONS, 3, 24, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Altered Microglia in the Amygdala Are Involved in Anxiety-related Behaviors of a Copy Number Variation Mouse Model of Autism

  Tomoko Shigemori, Atsushi Sakai, Toru Takumi, Yasuhiko Itoh, Hidenori Suzuki

  Apr. 2015, JOURNAL OF NIPPON MEDICAL SCHOOL, 82(2) (2), 92 - 99, English

  [Refereed]

  Scientific journal


• 3D visualization of the regional differences

  Ellegood, J., Anagnostou, E., Babineau, B.A., Crawley, J.N., Lin, L., Genestine, M., DiCicco-Bloom, E., Lai, J.K.Y., Foster, J.A., Pe{\~n}agarikano, O., Geschwind, D.H., Pacey, L.K., Hampson, D.R., Lalibert{\'e}, C.L., Mills, A.A., Tam, E., Osborne, L.R., Kouser, M., Espinosa-Becerra, F., Xuan, Z., Powell, C.M., Raznahan, A., Robins, D.M., Nakai, N., Nakatani, J., Takumi, T., van Eede, M.C., Kerr, T.M., Muller, C., Blakely, R.D., Veenstra-VanderWeele, J., Henkelman, R.M., Lerch, J.P.

  Feb. 2015, Molecular Psychiatry, 20(1) (1), 1, Co-authored internationally

  [Refereed]

  Scientific journal


• Clustering autism: Using neuroanatomical differences in 26 mouse models to gain insight into the heterogeneity

  Ellegood, J., Anagnostou, E., Babineau, B.A., Crawley, J.N., Lin, L., Genestine, M., Dicicco-Bloom, E., Lai, J.K.Y., Foster, J.A., Pe{\~n}agarikano, O., Geschwind, D.H., Pacey, L.K., Hampson, D.R., Lalibert{\'e}, C.L., Mills, A.A., Tam, E., Osborne, L.R., Kouser, M., Espinosa-Becerra, F., Xuan, Z., Powell, C.M., Raznahan, A., Robins, D.M., Nakai, N., Nakatani, J., Takumi, T., Van Eede, M.C., Kerr, T.M., Muller, C., Blakely, R.D., Veenstra-Vander Weele, J., Henkelman, R.M., Lerch, J.P.

  Feb. 2015, Molecular Psychiatry, 20(1) (1), 118 - 125, Co-authored internationally

  [Refereed]

  Scientific journal


• Towards understanding the neural mechanism of behavioral phenotypes seen in psychiatric disorders

  Nobuhiro Nakai, Ofer Yizhar, Toru Takumi

  Springer Japan, Jan. 2015, Optogenetics: Light-Sensing Proteins and their Applications, 331 - 340, English, Co-authored internationally

  [Refereed]

  In book


• Cerebellar plasticity and motor learning deficits in a copy-number variation mouse model of autism

  Claire Piochon, Alexander D. Kloth, Giorgio Grasselli, Heather K. Titley, Hisako Nakayama, Kouichi Hashimoto, Vivian Wan, Dana H. Simmons, Tahra Eissa, Jin Nakatani, Adriana Cherskov, Taisuke Miyazaki, Masahiko Watanabe, Toru Takumi, Masanobu Kano, Samuel S. -H. Wang, Christian Hansel

  Nov. 2014, NATURE COMMUNICATIONS, 5, 5586, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Genomic and genetic aspects of autism spectrum disorder

  Xiaoxi Liu, Toru Takumi

  Sep. 2014, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 452(2) (2), 244 - 253, English

  [Refereed]

  Scientific journal


• Enhanced synapse remodelling as a common phenotype in mouse models of autism

  Masaaki Isshiki, Shinji Tanaka, Toshihiko Kuriu, Katsuhiko Tabuchi, Toru Takumi, Shigeo Okabe

  Aug. 2014, NATURE COMMUNICATIONS, 5, 4742, English

  [Refereed]

  Scientific journal


• Developmental psychiatric disorders : Understanding genes, circuitries, and neurobiology using mouse models

  Takumi Toru, Sakurai Takesi

  It has been hypothesized that in neurodevelopmental psychiatric disorders such as schizophrenia and autism, biological processes in brain development may be affected by genetic and environmental causes, leading to alterations in brain wiring and/or synaptic formation and function, which manifest as characteristic phenotypes of these disorders. In fact, genetic studies have identified several genetic changes associated with schizophrenia and autism that would alter molecular pathways involved in brain development. It is also noted that the same genetic changes would lead to different clinical entities, such as autism and schizophrenia, emphasizing the importance of understanding basic neurobiology of brain wiring and synapse formation/function, in order to understand how functional alterations (and distinct phenotypes among psychiatric disorders) appear when those processes go away in these psychiatric disorders. To this end, we could take advantage of genetic findings in humans to develop mouse models with construct validity that have alterations in genes associated with these psychiatric disorders in humans and should gain insights into neurobiology underlying these psychiatric disorders by characterizing them.

  Japanese Society of Biological Psychiatry, 2014, Japanese Journal of Biological Psychiatry, 25(4) (4), 187 - 189, Japanese


• A Novel Protein, CHRONO, Functions as a Core Component of the Mammalian Circadian Clock

  Akihiro Goriki, Fumiyuki Hatanaka, Jihwan Myung, Jae Kyoung Kim, Takashi Yoritaka, Shintaro Tanoue, Takaya Abe, Hiroshi Kiyonari, Katsumi Fujimoto, Yukio Kato, Takashi Todo, Akio Matsubara, Daniel Forger, Toru Takumi

  Public Library of Science, 2014, PLoS Biology, 12(4) (4), e1001839, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Nuclear receptor-mediated cell-Autonomous oscillatory expression of the circadian transcription factor, neuronal pas domain protein 2 (NPAS2)

  Ritsuko Matsumura, Chiaki Matsubara, Koichi Node, Toru Takumi, Makoto Akashi

  Dec. 2013, Journal of Biological Chemistry, 288(51) (51), 36548 - 36553, English

  [Refereed]

  Scientific journal


• Helix-loop-helix protein Id2 stabilizes mammalian circadian oscillation under constant light conditions.

  Akihito A Adachi, Atsuko Fujioka, Mamoru Nagano, Koh-hei Masumoto, Toru Takumi, Takashi Yoshimura, Shizufumi Ebihara, Kentaro Mori, Yoshifumi Yokota, Yasufumi Shigeyoshi

  The mammalian circadian oscillator is composed of interacting positive and negative transcription events. The clock proteins PER1 and PER2 play essential roles in a negative limb of the feedback loop that generates the circadian rhythm in mammals. In addition, the proteins CLOCK and BMAL1 (also known as ARNTL) form a heterodimer that drives the Per genes via the E-box consensus sequences within their promoter regions. In the present study, we demonstrate that Id2 is involved in stabilization of the amplitudes of the circadian oscillations by suppressing transcriptional activation of clock genes Clock and Bmal1. Id2 shows dynamic oscillation in the SCN, with a peak in the late subjective night. Under constant dark conditions (DD), Id2(-/-) mice showed no apparent difference in locomotor activity, however, under constant light conditions (LL), Id2(-/-) mice exhibit aberrant locomotor activity, with lower circadian oscillation amplitudes, although the free running periods in Id2(-/-) mice show no differences from those in either wild type or heterozygous mice. Id2(-/-) animals also exhibit upregulation of Per1 in constant light, during both the subjective night and day. In wild type mice, Id2 is upregulated by constant light exposure during the subjective night. We propose that Id2 expression in the SCN contributes to maintenance of dynamic circadian oscillations.

  Dec. 2013, Zoological science, 30(12) (12), 1011 - 8, English, Domestic magazine

  [Refereed]

  Scientific journal


• Characterization and Modeling of Intermittent Locomotor Dynamics in Clock Gene-Deficient Mice

  Toru Nakamura, Toru Takumi, Atsuko Takano, Fumiyuki Hatanaka, Yoshiharu Yamamoto

  Mar. 2013, PLOS ONE, 8(3) (3), e58884, English

  [Refereed]

  Scientific journal


• A 15q11-q13 Duplication Mouse Model of Autism Spectrum Disorders

  Toru Takumi, Keita Fukumoto, Jun Nomura

  Elsevier Inc., 2013, The Neuroscience of Autism Spectrum Disorders, 401 - 408, English

  [Refereed]

  In book


• Regulation of Zipcode Binding Protein 1 Transport Dynamics in Axons by Myosin Va

  Vijayalaxmi C. Nalavadi, Laura E. Griffin, Philip Picard-Fraser, Andrew M. Swanson, Toru Takumi, Gary J. Bassell

  Oct. 2012, JOURNAL OF NEUROSCIENCE, 32(43) (43), 15133 - 15141, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Altered Serotonin, Dopamine and Norepinepherine Levels in 15q Duplication and Angelman Syndrome Mouse Models

  M. Febin Farook, Michael DeCuypere, Keith Hyland, Toru Takumi, Mark S. LeDoux, Lawrence T. Reiter

  Aug. 2012, PLOS ONE, 7(8) (8), e43030, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Widespread binding of FUS along nascent RNA regulates alternative splicing in the brain

  Boris Rogelj, Laura E. Easton, Gireesh K. Bogu, Lawrence W. Stanton, Gregor Rot, Tomaz Curk, Blaz Zupan, Yoichiro Sugimoto, Miha Modic, Nejc Haberman, James Tollervey, Ritsuko Fujii, Toru Takumi, Christopher E. Shaw, Jernej Ule

  Aug. 2012, SCIENTIFIC REPORTS, 2, 603, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Period Coding of Bmal1 Oscillators in the Suprachiasmatic Nucleus

  Jihwan Myung, Sungho Hong, Fumiyuki Hatanaka, Yoshihiro Nakajima, Erik De Schutter, Toru Takumi

  Jun. 2012, JOURNAL OF NEUROSCIENCE, 32(26) (26), 8900 - 8918, English

  [Refereed]

  Scientific journal


• REV-ERBα and the clock gene machinery in mouse peripheral tissues: A possible role as a synchronizing hinge

  Gianluigi Mazzoccoli, Y. Cai, S. Liu, M. Francavilla, F. Giuliani, A. Piepoli, V. Pazienza, M. Vinciguerra, T. Yamamoto, T. Takumi

  Apr. 2012, Journal of Biological Regulators and Homeostatic Agents, 26(2) (2), 265 - 276, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Mapping genes related to early onset major depressive disorder in dagestan genetic isolates

  Bulayeva, K., Lencz, T., Glatt, S., Takumi, T., Gurgenova, F., Kawakami, H., Bulayev, O.

  2012, Turk Psikiyatri Dergisi, 23(3) (3)

  Scientific journal


• Animal models of psychiatric disorDers that reflect human copy number variation

  Jun Nomura, Toru Takumi

  Hindawi Publishing Corporation, 2012, Neural Plasticity, 2012, 589524, English

  [Refereed]


• Differential Patterns in the Periodicity and Dynamics of Clock Gene Expression in Mouse Liver and Stomach

  Gianluigi Mazzoccoli, Massimo Francavilla, Valerio Pazienza, Giorgia Benegiamo, Ada Piepoli, Manlio Vinciguerra, Francesco Giuliani, Takuro Yamamoto, Toru Takumi

  2012, CHRONOBIOLOGY INTERNATIONAL, 29(10) (10), 1300 - 1311, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Genome-wide linkage scan of major depressive disorder in two dagestan genetic isolates

  Kazima B. Bulayeva, Todd Lencz, Stephen Glatt, Toru Takumi, Farida R. Gurgenova, Oleg A. Bulayev

  Oct. 2011, Central European Journal of Medicine, 6(5) (5), 616 - 624, English, Co-authored internationally

  [Refereed]

  Scientific journal


• The neurobiology of mouse models syntenic to human chromosome 15q

  Toru Takumi

  Sep. 2011, JOURNAL OF NEURODEVELOPMENTAL DISORDERS, 3(3) (3), 270 - 281, English

  [Refereed]

  Scientific journal


• The period of the somite segmentation clock is sensitive to Notch activity

  Woong Kim, Takaaki Matsui, Masataka Yamao, Makoto Ishibashi, Kota Tamada, Toru Takumi, Kenji Kohno, Shigeyuki Oba, Shin Ishii, Yuichi Sakumura, Yasumasa Bessho

  Sep. 2011, MOLECULAR BIOLOGY OF THE CELL, 22(18) (18), 3541 - 3549, English

  [Refereed]

  Scientific journal


• Genome-Wide Profiling of the Core Clock Protein BMAL1 Targets Reveals a Strict Relationship with Metabolism

  Fumiyuki Hatanaka, Chiaki Matsubara, Jihwan Myung, Takashi Yoritaka, Naoko Kamimura, Shuichi Tsutsumi, Akinori Kanai, Yutaka Suzuki, Paolo Sassone-Corsi, Hiroyuki Aburatani, Sumio Sugano, Toru Takumi

  Dec. 2010, MOLECULAR AND CELLULAR BIOLOGY, 30(24) (24), 5636 - 5648, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Decreased Exploratory Activity in a Mouse Model of 15q Duplication Syndrome; Implications for Disturbance of Serotonin Signaling

  Kota Tamada, Shozo Tomonaga, Fumiyuki Hatanaka, Nobuhiro Nakai, Keizo Takao, Tsuyoshi Miyakawa, Jin Nakatani, Toru Takumi

  Dec. 2010, PLOS ONE, 5(12) (12), e15126, English

  [Refereed]

  Scientific journal


• A humanoid mouse model of autism

  Toru Takumi

  Oct. 2010, BRAIN & DEVELOPMENT, 32(9) (9), 753 - 758, English

  [Refereed]

  Scientific journal


• Dual-Color Luciferase Mouse Directly Demonstrates Coupled Expression of Two Clock Genes

  Takako Noguchi, Tomoko Michihata, Wataru Nakamura, Toru Takumi, Ritsuko Shimizu, Masayuki Yamamoto, Masaaki Ikeda, Yoshihiro Ohmiya, Yoshihiro Nakajima

  Sep. 2010, BIOCHEMISTRY, 49(37) (37), 8053 - 8061, English

  [Refereed]

  Scientific journal


• Mutations of optineurin in amyotrophic lateral sclerosis.

  Hirofumi Maruyama, Hiroyuki Morino, Hidefumi Ito, Yuishin Izumi, Hidemasa Kato, Yasuhito Watanabe, Yoshimi Kinoshita, Masaki Kamada, Hiroyuki Nodera, Hidenori Suzuki, Osamu Komure, Shinya Matsuura, Keitaro Kobatake, Nobutoshi Morimoto, Koji Abe, Naoki Suzuki, Masashi Aoki, Akihiro Kawata, Takeshi Hirai, Takeo Kato, Kazumasa Ogasawara, Asao Hirano, Toru Takumi, Hirofumi Kusaka, Koichi Hagiwara, Ryuji Kaji, Hideshi Kawakami

  Amyotrophic lateral sclerosis (ALS) has its onset in middle age and is a progressive disorder characterized by degeneration of motor neurons of the primary motor cortex, brainstem and spinal cord. Most cases of ALS are sporadic, but about 10% are familial. Genes known to cause classic familial ALS (FALS) are superoxide dismutase 1 (SOD1), ANG encoding angiogenin, TARDP encoding transactive response (TAR) DNA-binding protein TDP-43 (ref. 4) and fused in sarcoma/translated in liposarcoma (FUS, also known as TLS). However, these genetic defects occur in only about 20-30% of cases of FALS, and most genes causing FALS are unknown. Here we show that there are mutations in the gene encoding optineurin (OPTN), earlier reported to be a causative gene of primary open-angle glaucoma (POAG), in patients with ALS. We found three types of mutation of OPTN: a homozygous deletion of exon 5, a homozygous Q398X nonsense mutation and a heterozygous E478G missense mutation within its ubiquitin-binding domain. Analysis of cell transfection showed that the nonsense and missense mutations of OPTN abolished the inhibition of activation of nuclear factor kappa B (NF-kappaB), and the E478G mutation revealed a cytoplasmic distribution different from that of the wild type or a POAG mutation. A case with the E478G mutation showed OPTN-immunoreactive cytoplasmic inclusions. Furthermore, TDP-43- or SOD1-positive inclusions of sporadic and SOD1 cases of ALS were also noticeably immunolabelled by anti-OPTN antibodies. Our findings strongly suggest that OPTN is involved in the pathogenesis of ALS. They also indicate that NF-kappaB inhibitors could be used to treat ALS and that transgenic mice bearing various mutations of OPTN will be relevant in developing new drugs for this disorder.

  7295, May 2010, Nature, 465(7295) (7295), 223 - 6, English, International magazine

  [Refereed]

  Scientific journal


• A protein-protein interaction of stress-responsive myosin VI endowed to inhibit neural progenitor self-replication with RNA binding protein, TLS, in murine hippocampus

  Takeshi Takarada, Keisuke Tamaki, Toru Takumi, Masato Ogura, Yuma Ito, Noritaka Nakamichi, Yukio Yoneda

  Sep. 2009, JOURNAL OF NEUROCHEMISTRY, 110(5) (5), 1457 - 1468, English

  [Refereed]

  Scientific journal


• Autonomous regulation of osteosarcoma cell invasiveness by Wnt5a/Ror2 signaling

  M. Enomoto, S. Hayakawa, S. Itsukushima, D. Y. Ren, M. Matsuo, K. Tamada, C. Oneyama, M. Okada, T. Takumi, M. Nishita, Y. Minami

  Sep. 2009, ONCOGENE, 28(36) (36), 3197 - 3208, English

  [Refereed]

  Scientific journal


• Fezf1 Is Required for Penetration of the Basal Lamina by Olfactory Axons to Promote Olfactory Development

  Yasuhito Watanabe, Kiyoshi Inoue, Ayako Okuyama-Yamamoto, Nobuhiro Nakai, Jin Nakatani, Ken-Ichi Nibu, Naoko Sato, Yasuhiko Iiboshi, Kosuke Yusa, Gen Kondoh, Junji Takeda, Toshio Terashima, Toru Takumi

  Aug. 2009, JOURNAL OF COMPARATIVE NEUROLOGY, 515(5) (5), 565 - 584, English

  [Refereed]

  Scientific journal


• Abnormal Behavior in a Chromosome-Engineered Mouse Model for Human 15q11-13 Duplication Seen in Autism

  Jin Nakatani, Kota Tamada, Fumiyuki Hatanaka, Satoko Ise, Hisashi Ohta, Kiyoshi Inoue, Shozo Tomonaga, Yasuhito Watanabe, Yeun Jun Chung, Ruby Banerjee, Kazuya Iwamoto, Tadafumi Kato, Makoto Okazawa, Kenta Yamauchi, Koichi Tanda, Keizo Takao, Tsuyoshi Miyakawa, Allan Bradley, Toru Takumi

  Jun. 2009, CELL, 137(7) (7), 1235 - 1246, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Robust Food Anticipatory Activity in BMAL1-Deficient Mice

  Julie S. Pendergast, Wataru Nakamura, Rio C. Friday, Fumiyuki Hatanaka, Toru Takumi, Shin Yamazaki

  Mar. 2009, PLOS ONE, 4(3) (3), e4860, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Time microscopy of circadian expression of Cardiac Clock Gene mRNA Transcription: Chronodiagnostic and Chronotherapeutic implications

  R. B. Sothern, G. Cornelissen, T. Yamamoto, T. Takumi, F. Halberg

  Mar. 2009, CLINICA TERAPEUTICA, 160(2) (2), E25 - E34, English, Co-authored internationally

  [Refereed]


• TLS-GFP cannot rescue mRNP formation near spines and spine phenotype in TLS-KO

  Ritsuko Fujii, Olga Grossenbacher-Zinchuk, Ildasolha Jamari, Yu Wang, Vadim Zinchuk, Toru Takumi

  Jan. 2009, NEUROREPORT, 20(1) (1), 57 - 61, English, Co-authored internationally

  [Refereed]

  Scientific journal


• TLS interaction with NMDA R1 splice variant in retinal ganglion cell line RGC-5

  Widyawilis Selamat, Ildasolha Jamari, Yu Wang, Toru Takumi, Fulton Wong, Ristuko Fujii

  Jan. 2009, NEUROSCIENCE LETTERS, 450(2) (2), 163 - 166, English, Co-authored internationally

  [Refereed]

  Scientific journal


• The resetting of the circadian rhythm by Prostaglandin J(2) is distinctly phase-dependent

  Satoshi Koinuma, Kazuhiro Yagita, Atsuko Fujioka, Naoyuki Takashima, Toru Takumi, Yasufumi Shigeyoshi

  Jan. 2009, FEBS LETTERS, 583(2) (2), 413 - 418, English

  [Refereed]

  Scientific journal


• Central and peripheral circadian clock genes, their statistical analysis for rhythms, and relationship to health and disease

  Sothern, R.B., Yamamoto, T., Corn{\'e}lissen, G., Takumi, T., Halberg, F.

  2009, Scripta Medica Facultatis Medicae Universitatis Brunensis Masarykianae, 82(3) (3)

  Scientific journal


• Comment on "Differential rescue of light- and food-entrainable circadian rhythms".

  Mistlberger RE, Yamazaki S, Pendergast JS, Landry GJ, Takumi T, Nakamura W

  Oct. 2008, Science (New York, N.Y.), 322(5902) (5902), 675; author reply 675, English, Co-authored internationally

  [Refereed]


• Of Mice and Men - Universality and Breakdown of Behavioral Organization

  Toru Nakamura, Toru Takumi, Atsuko Takano, Naoko Aoyagi, Kazuhiro Yoshiuchi, Zbigniew R. Struzik, Yoshiharu Yamamoto

  Apr. 2008, PLOS ONE, 3(4) (4), e2050, English

  [Refereed]

  Scientific journal


• In vivo monitoring of circadian timing in freely moving mice

  Wataru Nakamura, Shin Yamazaki, Takahiro J. Nakamura, Tetsuo Shirakawa, Gene D. Block, Toru Takumi

  Mar. 2008, CURRENT BIOLOGY, 18(5) (5), 381 - 385, English, Co-authored internationally

  [Refereed]

  Scientific journal


• A direct repeat of E-box-like elements is required for cell-autonomous circadian rhythm of clock genes

  Yasukazu Nakahata, Mayumi Yoshida, Atsuko Takano, Haruhiko Soma, Takuro Yamamoto, Akio Yasuda, Toru Nakatsu, Toru Takumi

  Jan. 2008, BMC MOLECULAR BIOLOGY, 9, 1, English

  [Refereed]

  Scientific journal


• Myosin-Va facilitates the accumulation of mRNA/protein complex in dendritic spines

  Atsushi Yoshimura, Ritsuko Fujii, Yasuhito Watanabe, Shigeo Okabe, Kenji Fukui, Toru Takumi

  Dec. 2006, CURRENT BIOLOGY, 16(23) (23), 2345 - 2351, English

  [Refereed]

  Scientific journal


• Molecular mechanism of cell-autonomous circadian gene expression of Period2, a crucial regulator of the mammalian circadian clock

  M Akashi, T Ichise, T Mamine, T Takumi

  Feb. 2006, MOLECULAR BIOLOGY OF THE CELL, 17(2) (2), 555 - 565, English

  [Refereed]

  Scientific journal


• The in vitro real-time oscillation monitoring system identifies potential entrainment factors for circadian clocks

  Y Nakahata, M Akashi, D Trcka, A Yasuda, T Takumi

  Feb. 2006, BMC MOLECULAR BIOLOGY, 7, 5, English

  [Refereed]

  Scientific journal


• Transcriptional oscillatory mechanism of molecular clocks

  Takumi, T.

  2006, Seikagaku the Journal of Japanese Biochemical Society, 78(5) (5)

  Scientific journal


• Biological anomalies of mental diseases are located in spines?

  Takumi, T.

  2006, Tanpakushitsu Kakusan Koso Protein Nucleic Acid Enzyme, 51(15) (15)

  Scientific journal


• Acute physical stress elevates mouse Period1 mRNA expression in mouse peripheral tissues via a glucocorticoid-responsive element

  T Yamamoto, Y Nakahata, M Tanaka, M Yoshida, H Soma, K Shinohara, A Yasuda, T Mamine, T Takumi

  Dec. 2005, JOURNAL OF BIOLOGICAL CHEMISTRY, 280(51) (51), 42036 - 42043, English

  [Refereed]

  Scientific journal


• TLS facilitates transport of mRNA encoding an actin-stabilizing protein to dendritic spines

  R Fujii, T Takumi

  Dec. 2005, JOURNAL OF CELL SCIENCE, 118(24) (24), 5755 - 5765, English

  [Refereed]

  Scientific journal


• The orphan nuclear receptor ROR alpha regulates circadian transcription of the mammalian core-clock Bmal1

  M Akashi, T Takumi

  May 2005, NATURE STRUCTURAL & MOLECULAR BIOLOGY, 12(5) (5), 441 - 448, English

  [Refereed]

  Scientific journal


• Importin alpha/beta mediates nuclear transport of a mammalian circadian clock component, mCRY2, together with mPER2, through a bipartite nuclear localization signal

  Y Sakakida, Y Miyamoto, E Nagoshi, M Akashi, TJ Nakamura, T Mamine, M Kasahara, Y Minami, Y Yoneda, T Takumi

  Apr. 2005, JOURNAL OF BIOLOGICAL CHEMISTRY, 280(14) (14), 13272 - 13278, English

  [Refereed]

  Scientific journal


• The RNA binding protein TLS is translocated to dendritic spines by mGluR5 activation and regulates spine morphology

  R Fujii, S Okabe, T Urushido, K Inoue, A Yoshimura, T Tachibana, T Nishikawa, GG Hicks, T Takumi

  Mar. 2005, CURRENT BIOLOGY, 15(6) (6), 587 - 593, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Fez1 is layer-specifically expressed in the adult mouse neocortex

  K Inoue, T Terashima, T Nishikawa, T Takumi

  Dec. 2004, EUROPEAN JOURNAL OF NEUROSCIENCE, 20(11) (11), 2909 - 2916, English

  [Refereed]

  Scientific journal


• The receptor tyrosine kinase Ror2 associates with and is activated by casein kinase Iepsilon.

  Shuichi Kani, Isao Oishi, Hiroyuki Yamamoto, Akinori Yoda, Hiroaki Suzuki, Akira Nomachi, Kengo Iozumi, Michiru Nishita, Akira Kikuchi, Toru Takumi, Yasuhiro Minami

  Ror2, a member of the mammalian Ror family of receptor tyrosine kinases, plays important roles in developmental morphogenesis, although the mechanism underlying activation of Ror2 remains largely elusive. We show that when expressed in mammalian cells, Ror2 associates with casein kinase Iepsilon (CKIepsilon), a crucial regulator of Wnt signaling. This association occurs primarily via the cytoplasmic C-terminal proline-rich domain of Ror2. We also show that Ror2 is phosphorylated by CKIepsilon on serine/threonine residues, in its C-terminal serine/threonine-rich 2 domain, resulting in autophosphorylation of Ror2 on tyrosine residues. Furthermore, it was found that association of Ror2 with CKIepsilon is required for its serine/threonine phosphorylation by CKIepsilon. Site-directed mutagenesis of tyrosine residues in Ror2 reveals that the sites of phosphorylation are contained among the five tyrosine residues in the proline-rich domain but not among the four tyrosine residues in the tyrosine kinase domain. Moreover, we show that in mammalian cells, CKIepsilon-mediated phosphorylation of Ror2 on serine/threonine and tyrosine residues is followed by the tyrosine phosphorylation of G protein-coupled receptor kinase 2, a kinase with a developmental expression pattern that is remarkably similar to that of Ror2. Intriguingly, a mutant of Ror2 lacking five tyrosine residues, including the autophosphorylation sites, fails to tyrosine phosphorylate G protein-coupled receptor kinase 2. This indicates that autophosphorylation of Ror2 is required for full activation of its tyrosine kinase activity. These findings demonstrate a novel role for CKIepsilon in the regulation of Ror2 tyrosine kinase.

  48, Nov. 2004, The Journal of biological chemistry, 279(48) (48), 50102 - 9, English, International magazine

  [Refereed]

  Scientific journal


• Domain Architectures and characterization of an RNA-binding protein, TLS

  Y Iko, TS Kodama, N Kasai, T Oyama, EH Morita, T Muto, M Okumura, R Fujii, T Takumi, S Tate, K Morikawa

  Oct. 2004, JOURNAL OF BIOLOGICAL CHEMISTRY, 279(43) (43), 44834 - 44840, English

  [Refereed]

  Scientific journal


• Transcriptional oscillation of canonical clock genes in mouse peripheral tissues

  T Yamamoto, Y Nakahata, H Soma, M Akashi, T Mamine, T Takumi

  Oct. 2004, BMC MOLECULAR BIOLOGY, 5, 18, English

  [Refereed]

  Scientific journal


• Synapse-associated protein 90/postsynaptic density-95-associated protein (SAPAP) is expressed differentially in phencyclidine-treated rats and is increased in the nucleus accumbens of patients with schizophrenia

  Y Kajimoto, O Shirakawa, XH Lin, T Hashimoto, N Kitamura, N Murakami, T Takumi, K Maeda

  Oct. 2003, NEUROPSYCHOPHARMACOLOGY, 28(10) (10), 1831 - 1839, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Regulation by gonadal steroids of the mRNA encoding for a type I receptor for TGF-beta in the female rat hypothalamus

  S Bouret, Prevot, V, T Takumi, JC Beauvillain, Mitchell, V

  Jul. 2002, NEUROENDOCRINOLOGY, 76(1) (1), 1 - 7, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Restoration of circadian behavioural rhythms in a period null Drosophila mutant (per(01)) by mammalian period homologues mPer1 and mPer2

  Y Shigeyoshi, E Meyer-Bernstein, K Yagita, WL Fu, YF Chen, T Takumi, P Schotland, A Sehgal, H Okamura

  Feb. 2002, GENES TO CELLS, 7(2) (2), 163 - 171, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Rapid cDNA cloning by PCR screening (RC-PCR).

  Takumi, T.

  2002, Methods in Molecular Biology Clifton N J, 192, 385 - 389

  [Refereed]

  Scientific journal


• Evidence that TGF beta may directly modulate POMC mRNA expression in the female rat arcuate nucleus

  S Bouret, MT Van Chuoi-Mariot, Prevot, V, D Croix, T Takumi, S Jegou, H Vaudry, JC Beauvillain, Mitchell, V

  Sep. 2001, ENDOCRINOLOGY, 142(9) (9), 4055 - 4065, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Inhibition of cardiac delayed rectifier K+ currents by an antisense oligodeoxynucleotide against IsK (minK) and over-expression of IsK mutant D77N in neonatal mouse hearts

  H Ohyama, H Kajita, K Omori, T Takumi, N Hiramoto, T Iwasaka, H Matsuda

  Jun. 2001, PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY, 442(3) (3), 329 - 335, English

  [Refereed]

  Scientific journal


• Novel nonsense mutation in the Na+/HCO3- cotransporter gene (SLC4A4) in a patient with permanent isolated proximal renal tubular acidosis and bilateral glaucoma

  Igarashi, T., Inatomi, J., Sekine, T., Seki, G., Shimadzu, M., Tozawa, F., Takeshima, Y., Takumi, T., Takahashi, T., Yoshikawa, N., Nakamura, H., Endou, H.

  2001, Journal of the American Society of Nephrology, 12(4) (4)

  Scientific journal


• Evidence that members of the TGF beta superfamily play a role in regulation of the GnRH neuroendocrine axis: Expression of a type I serine-threonine kinase receptor for TGR beta and activin in GnRH neurones and hypothalamic areas of the female rat

  Prevot, V, S Bouret, D Croix, T Takumi, L Jennes, Mitchell, V, JC Beauvillain

  Jul. 2000, JOURNAL OF NEUROENDOCRINOLOGY, 12(7) (7), 665 - 670, English, Co-authored internationally

  [Refereed]

  Scientific journal


• A putative transcription factor with seven zinc-finger motifs identified in the developing suprachiasmatic nucleus by the differential display PCR method

  Y Maebayashi, Y Shigeyoshi, T Takumi, H Okamura

  Nov. 1999, JOURNAL OF NEUROSCIENCE, 19(22) (22), 10176 - 10183, English

  [Refereed]

  Scientific journal


• A mammalian ortholog of Drosophila timeless, highly expressed in SCN and retina, forms a complex with mPER1

  T Takumi, Y Nagamine, S Miyake, C Matsubara, K Taguchi, S Takekida, Y Sakakida, K Nishikawa, T Kishimoto, S Niwa, K Okumura, H Okamura

  Jan. 1999, GENES TO CELLS, 4(1) (1), 67 - 75, English

  [Refereed]

  Scientific journal


• A light-independent oscillatory gene mPer3 in mouse SCN and OVLT

  T Takumi, K Taguchi, S Miyake, Y Sakakida, N Takashima, C Matsubara, Y Maebayashi, K Okumura, S Takekida, S Yamamoto, K Yagita, L Yan, MW Young, H Okamura

  Aug. 1998, EMBO JOURNAL, 17(16) (16), 4753 - 4759, English, Co-authored internationally

  [Refereed]

  Scientific journal


• New mammalian period gene predominantly expressed in the suprachiasmatic nucleus

  T Takumi, C Matsubara, Y Shigeyoshi, K Taguchi, K Yagita, Y Maebayashi, Y Sakakida, K Okumura, N Takashima, H Okamura

  Mar. 1998, GENES TO CELLS, 3(3) (3), 167 - 176, English

  [Refereed]

  Scientific journal


• Assignment of the glial inwardly rectifying potassium channel K-AB-2/Kir4.1 (Kcnj10) gene to the distal region of mouse chromosome 1

  Y Tada, Y Horio, T Takumi, M Terayama, L Tsuji, NG Copeland, NA Jenkins, Y Kurachi

  Nov. 1997, GENOMICS, 45(3) (3), 629 - 630, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Identification of novel homologues of mouse importin alpha, the alpha subunit of the nuclear pore-targeting complex, and their tissue-specific expression

  L Tsuji, T Takumi, N Imamoto, Y Yoneda

  Oct. 1997, FEBS LETTERS, 416(1) (1), 30 - 34, English

  [Refereed]

  Scientific journal


• A dye terminator method for automated DNA sequencing using four fluorescent dideoxynucleosides and thermal cycling

  T Takumi, H Fujiwake, Y Kurachi

  Oct. 1997, ANALYTICAL SCIENCES, 13(5) (5), 735 - 739, English

  [Refereed]

  Scientific journal


• Assignment of the murine inwardly rectifying potassium channel IRK3 gene (Kcnj4) to the mouse Chromosome 15

  K Morishige, T Takumi, N Takahashi, H Koyama, H Kurachi, A Miyake, Y Murata, NG Copeland, DJ Gilbert, NA Jenkins, Y Kurachi

  Sep. 1997, MAMMALIAN GENOME, 8(9) (9), 699 - 700, English, Co-authored internationally

  [Refereed]

  Scientific journal


• Use of PCR for cDNA library screening.

  Takumi, T.

  1997, Methods in Molecular Biology Clifton N J, 67, 339 - 344

  [Refereed]

  Scientific journal


• Distinct localization of two serine-threonine kinase receptors for activin and TGF-beta in the rat brain and down-regulation of type I activin receptor during peripheral nerve regeneration

  N Morita, T Takumi, H Kiyama

  Dec. 1996, MOLECULAR BRAIN RESEARCH, 42(2) (2), 263 - 271, English

  [Refereed]

  Scientific journal


• Assignment of the murine inward rectifier potassium channel Irk2 (Kir2.2) gene to the central region of mouse chromosome 11

  T Takumi, L Tsuji, C Kondo, N Takahashi, K Morishige, NG Copeland, DJ Gilbert, NA Jenkins, Y Kurachi

  Oct. 1996, GENOMICS, 37(2) (2), 270 - 272, English, Co-authored internationally

  [Refereed]

  Scientific journal


• A novel ubiquitously distributed isoform of GIRK2 (GIRK2B) enhances GIRK1 expression of the G-protein-gated K+ current in Xenopus oocytes

  S Isomoto, C Kondo, N Takahashi, S Matsumoto, M Yamada, T Takumi, Y Horio, Y Kurachi

  Jan. 1996, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 218(1) (1), 286 - 291, English

  [Refereed]

  Scientific journal


• A NOVEL ATP-DEPENDENT INWARD RECTIFIER POTASSIUM CHANNEL EXPRESSED PREDOMINANTLY IN GLIAL-CELLS

  T TAKUMI, T ISHII, Y HORIO, K MORISHIGE, N TAKAHASHI, M YAMADA, T YAMASHITA, H KIYAMA, K SOHMIYA, S NAKANISHI, Y KURACHI

  Jul. 1995, JOURNAL OF BIOLOGICAL CHEMISTRY, 270(27) (27), 16339 - 16346, English

  [Refereed]

  Scientific journal


• G(BETA-GAMMA) DIRECTLY BINDS TO THE CARBOXYL-TERMINUS OF THE G-PROTEIN-GATED MUSCARINIC K+ CHANNEL, GIRK1

  A INANOBE, KI MORISHIGE, N TAKAHASHI, H ITO, M YAMADA, T TAKUMI, H NISHINA, K TAKAHASHI, Y KANAHO, T KATADA, Y KURACHI

  Jul. 1995, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 212(3) (3), 1022 - 1028, English

  [Refereed]

  Scientific journal


• MOLECULAR-BASIS OF I-SK PROTEIN-REGULATION BY OXIDATION OR CHELATION

  AE BUSCH, S WALDEGGER, T HERZER, G RABER, E GULBINS, T TAKUMI, H MORIYOSHI, S NAKANISHI, F LANG

  Feb. 1995, JOURNAL OF BIOLOGICAL CHEMISTRY, 270(8) (8), 3638 - 3641, English, Co-authored internationally

  [Refereed]

  Scientific journal


• MOLECULAR CHARACTERIZATION OF A TYPE-I SERINE THREONINE KINASE RECEPTOR FOR TGF-BETA AND ACTIVIN IN THE RAT PITUITARY-TUMOR CELL-LINE GH3

  T TAKUMI, A MOUSTAKAS, HY LIN, HF LODISH

  Jan. 1995, EXPERIMENTAL CELL RESEARCH, 216(1) (1), 208 - 214, English, Co-authored internationally

  [Refereed]

  Scientific journal


• GH3 PITUITARY-TUMOR CELLS CONTAIN HETEROMERIC TYPE-I AND TYPE-II RECEPTOR COMPLEXES FOR TRANSFORMING GROWTH-FACTOR-BETA AND ACTIVIN-A

  A MOUSTAKAS, T TAKUMI, HY LIN, HF LODISH

  Jan. 1995, JOURNAL OF BIOLOGICAL CHEMISTRY, 270(2) (2), 765 - 769, English, Co-authored internationally

  [Refereed]

  Scientific journal


• MOLECULAR AND FUNCTIONAL-HETEROGENEITY OF INWARD RECTIFIER POTASSIUM CHANNELS IN BRAIN AND HEART

  N TAKAHASHI, KI MORISHIGE, M YAMADA, T TAKUMI, Y KURACHI

  1995, HEART AND VESSELS, 8 - 11, English

  [Refereed]

  Scientific journal


• BLOCKADE OF HUMAN I-SK CHANNELS EXPRESSED IN XENOPUS OOCYTES BY THE NOVEL CLASS-III ANTIARRHYTHMIC NE-10064

  AE BUSCH, T HERZER, T TAKUMI, P KRIPPEITDREWS, S WALDEGGER, F LANG

  Oct. 1994, EUROPEAN JOURNAL OF PHARMACOLOGY, 264(1) (1), 33 - 37, English, Co-authored internationally

  [Refereed]

  Scientific journal


• RAPID CDNA CLONING BY PCR SCREENING

  T TAKUMI, HF LODISH

  Sep. 1994, BIOTECHNIQUES, 17(3) (3), 443 - 444, English, Co-authored internationally

  [Refereed]


• ALTERATIONS OF GATING PARAMETERS BY NEUTRAL SUBSTITUTIONS OF TRANSMEMBRANE LEU(52) OF SLOW POTASSIUM CHANNEL

  S OIKI, T TAKUMI, Y OKADA, S NAKANISHI

  1993, MOLECULAR BASIS OF ION CHANNELS AND RECEPTORS INVOLVED IN NERVE EXCITATION, SYNAPTIC TRANSMISSION AND MUSCLE CONTRACTION, 707, 402 - 406, English

  [Refereed]

  Scientific journal


• Amplification of ten deletion-rich exons of the dystrophin gene by polymerase chain reaction shows deletions in 36 of 90 Japanese families with Duchenne muscular dystrophy

  Kitoh, Y., Matsuo, M., Nishio, H., Takumi, T., Nakajima, T., Masumura, T., Koga, J., Nakamura, H.

  1992, American Journal of Medical Genetics, 42(4) (4)

  Scientific journal


• CELLULAR-LOCALIZATION OF RAT ISK PROTEIN IN THE STRIA VASCULARIS BY IMMUNOHISTOCHEMICAL OBSERVATION

  M SAKAGAMI, K FUKAZAWA, T MATSUNAGA, H FUJITA, N MORI, T TAKUMI, H OHKUBO, S NAKANISHI

  Nov. 1991, HEARING RESEARCH, 56(1-2) (1-2), 168 - 172, English

  [Refereed]

  Scientific journal


• ALTERATION OF CHANNEL ACTIVITIES AND GATING BY MUTATIONS OF SLOW-ISK POTASSIUM CHANNEL

  T TAKUMI, K MORIYOSHI, ARAMORI, I, T ISHII, S OIKI, Y OKADA, H OHKUBO, S NAKANISHI

  Nov. 1991, JOURNAL OF BIOLOGICAL CHEMISTRY, 266(33) (33), 22192 - 22198, English

  [Refereed]

  Scientific journal


• Molecular characterization of mammalian tachykinin receptors and a possible epithelial potassium channel

  Shigetada Nakanishi, Hiroaki Ohkubo, Akira Kakizuka, Yoshifumi Yokota, Ryuichi Shigemoto, Yoshiki Sasai, Toru Takumi

  1991, Recent Progress in Hormone Research, 46(1) (1), 59 - 83, English

  [Refereed]

  Scientific journal


• Exon skipping during splicing of dystrophin mRNA precursor due to an intraexon deletion in the dystrophin gene of Duchenne muscular dystrophy Kobe

  Matsuo, M., Masumura, T., Nishio, H., Nakajima, T., Kitoh, Y., Takumi, T., Koga, J., Nakamura, H.

  1991, Journal of Clinical Investigation, 87(6) (6)

  Scientific journal


• GENERATION OF TRANSGENIC MICE WITH ELEVATED BLOOD-PRESSURE BY INTRODUCTION OF THE RAT RENIN AND ANGIOTENSINOGEN GENES

  H OHKUBO, H KAWAKAMI, Y KAKEHI, T TAKUMI, H ARAI, Y YOKOTA, M IWAI, Y TANABE, M MASU, J HATA, H IWAO, H OKAMOTO, M YOKOYAMA, T NOMURA, M KATSUKI, S NAKANISHI

  Jul. 1990, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 87(13) (13), 5153 - 5157, English

  [Refereed]

  Scientific journal


• IMMUNOHISTOCHEMICAL STUDY OF A RAT MEMBRANE-PROTEIN WHICH INDUCES A SELECTIVE POTASSIUM PERMEATION - ITS LOCALIZATION IN THE APICAL MEMBRANE PORTION OF EPITHELIAL-CELLS

  T SUGIMOTO, Y TANABE, R SHIGEMOTO, M IWAI, T TAKUMI, H OHKUBO, S NAKANISHI

  Jan. 1990, JOURNAL OF MEMBRANE BIOLOGY, 113(1) (1), 39 - 47, English

  [Refereed]

  Scientific journal


• A very small frame-shifting deletion within exon 19 of the Duchenne muscular dystrophy gene

  Matsuo, M., Masumura, T., Nakajima, T., Kitoh, Y., Takumi, T., Nishio, H., Koga, J., Nakamura, H.

  1990, Biochemical and Biophysical Research Communications, 170(2) (2)

  Scientific journal


• MOLECULAR-CLONING AND SEQUENCE-ANALYSIS OF HUMAN GENOMIC DNA ENCODING A NOVEL MEMBRANE-PROTEIN WHICH EXHIBITS A SLOWLY ACTIVATING POTASSIUM CHANNEL ACTIVITY

  T MURAI, A KAKIZUKA, T TAKUMI, H OHKUBO, S NAKANISHI

  May 1989, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 161(1) (1), 176 - 181, English

  [Refereed]

  Scientific journal


• CLONING OF A MEMBRANE-PROTEIN THAT INDUCES A SLOW VOLTAGE-GATED POTASSIUM CURRENT

  T TAKUMI, H OHKUBO, S NAKANISHI

  Nov. 1988, SCIENCE, 242(4881) (4881), 1042 - 1045, English

  [Refereed]

  Scientific journal

• PERIOD2(PER2)のリン酸化スイッチによる気分・概日リズムの制御機構の解明

  白藤俊彦, 早田敦子, 早田敦子, 山脇洋輔, 今村聖路, 今村聖路, 竹田浩之, 東山繁樹, 内匠透, 内匠透, 内匠透, 内匠透

  2023, 日本薬理学雑誌, 158(Supplement) (Supplement)


• Identification of cell type-specific autism risk factors by single-cell, organoid studies

  Nomura J, Takumi T

  Jun. 2022, Japanese Journal of Biological Psychiatry, 33(2) (2), 48 - 52


• 【ストレスと神経系】ストレス・レジリエンス(ストレスに対する感受性と抵抗性をめぐって) 概日リズムに関わる時計遺伝子とストレス感受性・抵抗性

  早田 敦子, 内匠 透

  (株)中外医学社, Jun. 2021, Clinical Neuroscience, 39(6) (6), 780 - 783, Japanese


• 自閉症学 細胞・マウスモデルから病態に迫る

  内匠 透

  (一社)日本小児神経学会, May 2021, 脳と発達, 53(Suppl.) (Suppl.), S77 - S77, Japanese


• 【脳の発生とその異常】遺伝子異常 ASDにみられる遺伝子異常とその特徴

  藤間 秀平, 内匠 透

  (株)中外医学社, Dec. 2020, Clinical Neuroscience, 38(12) (12), 1556 - 1560, Japanese


• 最新の自閉症学 自閉スペクトラム症の病態理解を目指して

  内匠 透

  (一社)日本児童青年精神医学会, Oct. 2020, 日本児童青年精神医学会総会抄録集, 61回, S1 - 1, Japanese


• 発達期におけるシナプス後肥厚のリモデリング

  貝塚 剛志, 鈴木 健裕, 岸 憲幸, 岡野 栄之, 堂前 直, 内匠 透

  (公社)日本生化学会, Sep. 2020, 日本生化学会大会プログラム・講演要旨集, 93回, [1Z12 - 617)], Japanese


• 【自閉症学の魅力】自閉症基礎研究を加速させる新技術とその展望

  野村 淳, 内匠 透

  日本生物学的精神医学会, Jun. 2020, 日本生物学的精神医学会誌, 31(2) (2), 71 - 75, Japanese


• 新規シナプスタンパク質Aの解析

  吉田 智史, 貝塚 剛志, 内匠 透, 高橋 健造

  琉球医学会, 2020, 琉球医学会誌, 39(1-4) (1-4), 90 - 90, Japanese


• 【働く細胞(メンタル編)】脈絡叢と精神疾患

  今村 聖路, 内匠 透

  (有)科学評論社, Oct. 2019, 精神科, 35(4) (4), 362 - 368, Japanese


• 発達期におけるシナプス後肥厚のリモデリング

  貝塚 剛志, 鈴木 健裕, 堂前 直, 内匠 透

  日本電気泳動学会, Jul. 2019, 電気泳動, 63(Suppl.) (Suppl.), 244 - 244, Japanese


• 【自閉症学-自閉症・発達障害の病態解明に向けて】自閉症の動物モデル

  仲西 萌絵, 内匠 透

  医歯薬出版(株), Jan. 2019, 医学のあゆみ, 268(3) (3), 190 - 194, Japanese


• 自閉症フェノタイピングのためのin vitro基礎研究のトレンド

  野村淳, 内匠透

  医歯薬出版(株), 2019, 医学のあゆみ, 268(3) (3), 184 - 190, Japanese


• 【脳神経回路と高次脳機能 スクラップ&ビルドによる心の発達と脳疾患の謎を解く】(第3章)脳発達・再編と病気・障害 発達障害 自閉症の病態とシナプス動態を中心に

  内匠 透

  (株)羊土社, Aug. 2018, 実験医学, 36(12) (12), 2072 - 2075, Japanese


• 脳神経回路と精神生物学 神経機能と心の根源を探る Caイメージングと精神神経機能

  三浦 勇, 中井 信裕, 戸谷 豪志, 川俣 貴一, 内匠 透

  (株)医薬ジャーナル社, Apr. 2018, Clinical Calcium, 28(5) (5), 702 - 708, Japanese


• 【遺伝子点変異による精神神経疾患の発症機構解明】新規NLGN1自閉症変異とヒト型モデルマウス

  仲西 萌絵, 内匠 透

  日本生物学的精神医学会, Mar. 2018, 日本生物学的精神医学会誌, 29(1) (1), 7 - 11, Japanese


• コピー数多型の異常を原因とする自閉症スペクトラム障害に共通したメカニズムの同定

  山崎 真実, 内匠 透, 野村 淳, 高橋 健造

  琉球医学会, Dec. 2017, 琉球医学会誌, 36(1-2) (1-2), 47 - 47, Japanese


• 病態モデル動物からアプローチする発達障害の分子病態理解 クロマチンリモデリングの異常によって発症するASDの分子病態

  片山 雄太, 西山 正章, 昌子 浩孝, 大川 恭行, 川村 淳生, 佐藤 哲也, 須山 幹太, 内匠 透, 宮川 剛, 中山 敬一

  生命科学系学会合同年次大会運営事務局, Dec. 2017, 生命科学系学会合同年次大会, 2017年度, [1AW21 - 2], Japanese


• 病態モデル動物からアプローチする発達障害の分子病態理解 自閉症責任領域15q11-q13モデルマウスにおける原因遺伝子の探索

  玉田 紘太, 福本 景太, 戸谷 豪志, Ruf Sandra, Spitz Francois, 内匠 透

  生命科学系学会合同年次大会運営事務局, Dec. 2017, 生命科学系学会合同年次大会, 2017年度, [1AW21 - 4], Japanese


• 自閉スペクトラム症のCNVに基づくモデル神経細胞のトランスクリプトーム解析

  六峰 弘晃, Zuko Amila, 河野 掌, Shin Jay, Carninci Pierro, 三澤 日出巳, 野村 淳, 内匠 透

  生命科学系学会合同年次大会運営事務局, Dec. 2017, 生命科学系学会合同年次大会, 2017年度, [2P - 1069], Japanese


• 遺伝子点変異による精神神経疾患の発症機構解明 新規NLGN1自閉症変異とヒト型モデルマウス

  内匠 透

  日本生物学的精神医学会・日本神経精神薬理学会, Sep. 2017, 日本生物学的精神医学会・日本神経精神薬理学会合同年会プログラム・抄録集, 39回・47回, 82 - 82, Japanese


• 自閉症研究の最前線 基礎から臨床までの多面的なアプローチ 自閉症のモデル研究

  内匠 透, 中井 信裕

  東北医学会, Jun. 2017, 東北医学雑誌, 129(1) (1), 26 - 27, Japanese


• 疾患モデル動物を用いた精神医学研究の展望

  戸谷豪志, 玉田紘太, 三澤日出巳, 内匠透

  科学評論社, 01 Mar. 2017, 精神科, 30(3) (3), 221 - 266, Japanese

  [Invited]

  Introduction scientific journal


• 【時間生物学の新展開】精神疾患の時間生物学的考察

  戸谷 豪志, 福本 景太, 玉田 紘太, 三澤 日出巳, 内匠 透

  (公財)金原一郎記念医学医療振興財団, Dec. 2016, 生体の科学, 67(6) (6), 584 - 588, Japanese


• Neonatal fluoxetine restores sociability in a mouse model of autism

  M. Nagano, F. Saitow, T. Takumi, H. Suzuki

  Oct. 2016, EUROPEAN NEUROPSYCHOPHARMACOLOGY, 26, S272 - S273, English

  Summary international conference


• 【時間医学-最新の話題-】時計遺伝子Clock gene

  内匠 透

  アニムス編集委員会, Jul. 2016, アニムス, 21(3) (3), 18 - 20, Japanese


• 脳科学から見た自閉スペクトラム症 自閉スペクトラム症の生物学的理解

  内匠 透

  (一社)日本神経精神薬理学会, Jul. 2016, 日本神経精神薬理学会年会プログラム・抄録集, 46回, 42 - 42, Japanese


• 発達障害と相関する新規CNV(15q25.2-25.3)動物モデルの開発と解析(Role of the 15q25.2-25.3 CNV in developmental disorders: genetic and behavioral analyses)

  野村 淳, 神田 暁史, 外丸 祐介, 内匠 透

  (一社)日本神経精神薬理学会, Jul. 2016, 日本神経精神薬理学会年会プログラム・抄録集, 46回, 225 - 225, English


• Neuroligin 1における自閉症関連変異の同定と機能解析

  仲西 萌絵, 野村 淳, Liu Xiaoxi, Bucan Maja, 高橋 英機, 阿部 学, Zhou Li, 崎村 建司, 内匠 透

  (一社)日本神経精神薬理学会, Jul. 2016, 日本神経精神薬理学会年会プログラム・抄録集, 46回, 226 - 226, Japanese


• A novel mutation associated autism in Neuroligin1

  Moe Nakanishi, Takashi Arai, Maja Bucan, Xiao Ji, Xiaoxi Liu, Jun Nomura, Eiki Takahashi, Toru Takumi

  Jun. 2016, INTERNATIONAL JOURNAL OF NEUROPSYCHOPHARMACOLOGY, 19, 294 - 294, English

  Summary international conference


• 発達障害医療をめぐる最新の話題 発達障害の生物学的理解

  内匠 透

  (一社)日本小児神経学会, May 2016, 脳と発達, 48(Suppl.) (Suppl.), S140 - S140, Japanese


• 自閉症研究の最新事情 (脳疾患の科学)

  福田 邦宏, 福本 景太, 玉田 紘太, 内匠 透

  東京化学同人, May 2016, 現代化学 = Chemistry today, (542) (542), 35 - 38, Japanese


• 【こころの動物モデル-サカナからサルまで-】マウスを用いた自閉症の研究

  石井 宏茂, 中井 信裕, 坪井 貴司, 内匠 透

  (株)先端医学社, Apr. 2016, 分子精神医学, 16(2) (2), 91 - 96, Japanese


• アンジェルマン症候群モデルマウスの大脳皮質におけるRNA-seq解析

  六峰弘晃, 六峰弘晃, 二階堂愛, 内匠透, 三澤日出巳

  2016, 日本分子生物学会年会プログラム・要旨集(Web), 39th


• マウスを用いた自閉症の研究

  石井宏茂, 中井信裕, 坪井貴司, 内匠透

  2016, 分子精神医学, (16) (16), 19 - 24, Japanese

  [Invited]

  Introduction scientific journal


• Wnt5a-Ror2シグナルは繊毛タンパク質IFT20の発現誘導を介してがん細胞の浸潤を制御する

  西田 満, 西尾 忠, 紙崎 孝基, 王 志超, 榎本 真宏, 玉田 紘太, 内匠 透, Hsu Victor W., Pazour Gregory J., 南 康博

  (公社)日本生化学会, Dec. 2015, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 88回・38回, [4T16L - 08(3P0029)], Japanese


• NEAT1lncRNAの難溶性を考慮したパラスペックル構造因子の再定量と構造構築原理の探究

  中條岳志, 中川真一, 川口哲哉, 内匠透, 廣瀬哲郎

  (公社)日本生化学会, Dec. 2015, 日本生化学会大会(Web), 88th, 2T25P-08(2P0765) (WEB ONLY) - 08(2P0765)], Japanese


• 発達期におけるD-セリンの役割 Pick1ノックアウトマウスをモデルとして(Role for neonatal D-serine signaling: prevention of physiological deficits in adult Pick1 KO mice)

  野村 淳, ジャロペレ・ハナ, ルイス・イーストマン, ヌネアバデ・ペドロ, フッペゴウグ・フレデリック, キャッシュパジェ・タイラ, エミリアーニ・フランシェスコ, コンドウ・マリ, 古屋 亜佐子, ランデクサルガド・メリッサ, ヤブズ・アイハン, 神谷 篤, 内匠 透, ヒュガナー・リチャード, プレトニコフ・ミカエル, パトリシオ・オドネル, 澤 明

  日本生物学的精神医学会・日本神経精神薬理学会, Sep. 2015, 日本生物学的精神医学会・日本神経精神薬理学会合同年会プログラム・抄録集, 37回・45回, 174 - 174, English


• 【自閉症の生物学】ニューラルサーキットジェネティクスによる自閉症研究

  中井 信裕, 九里 信夫, 内匠 透

  (株)学研メディカル秀潤社, Apr. 2015, 細胞工学, 34(5) (5), 476 - 482, Japanese


• 【自閉症の生物学】自閉症と腸内細菌

  大内田 理佳, 内匠 透

  (株)学研メディカル秀潤社, Apr. 2015, 細胞工学, 34(5) (5), 483 - 486, Japanese


• 15q重複モデルマウスにおける脳内ドーパミン神経系の変化

  黒宮 里紗, 菅谷 望, 北村 陽二, 三輪 大輔, 小阪 孝史, 小川 数馬, 内匠 透, 柴 和弘

  (公社)日本薬学会, Mar. 2015, 日本薬学会年会要旨集, 135年会(3) (3), 192 - 192, Japanese


• 注目の遺伝子(第28回) 新たな時計遺伝子Chronoの機能

  畠中 史幸, 中尾 みのり, 内匠 透

  (株)先端医学社, Jan. 2015, 分子精神医学, 15(1) (1), 48 - 50, Japanese


• 自閉症スペクトラム障害の動物モデル

  岸本恵子, 野村淳, 内匠透

  (株)中外医学社, 2015, 月刊臨床神経科学, 33(2) (2), 201 - 205, Japanese


• 自閉症のエクソーム解析

  仲西萌絵, 野村淳, 内匠透

  2015, 細胞工学, 34(5) (5), 458 - 459


• Animal models of Autism

  Mutsumine H, Nomura J, Takumi T

  (有)科学評論社, 2015, Psychiatry, 27(4) (4), 271 - 276, Japanese

  [Refereed]

  Introduction scientific journal


• Animal model of ASD

  Kishimoto K, Nomura J, Takumi T

  (株)学研メディカル秀潤社, 2015, Cell Engineering, 34(5) (5), 460 - 465, Japanese

  [Refereed]


• Trial to develop animal models of Autism

  Shinmoto K, Nomura J, Takumi T

  日本生物学的精神医学会, 2015, Japanese Society of Biological Psychiatry, 26(2) (2), 81 - 85, Japanese

  [Refereed]


• Serotonin disturbance in mouse models of autism spectrum disorders

  Tamada, K., Takumi, T.

  Neuromethods, 2015, Neuromethods, 100, 239 - 262, English

  Book review


• 環境適応と時計シグナル 環境・ストレスへの応答系としての概日システム ストレスと概日リズム

  中尾 みのり, 内匠 透

  (一社)日本生理学会, Nov. 2014, 日本生理学雑誌, 76(6) (6), 125 - 126, Japanese


• INTEGRATIVE ANALYSIS ON NEUROPHARMACOLOGICAL EFFECTS OF HONEY

  M. Akanmu, F. Hatanaka, K. Tamada, T. Takumi

  Jul. 2014, BASIC & CLINICAL PHARMACOLOGY & TOXICOLOGY, 115, 129 - 129, English

  Summary international conference


• 【精神疾患の病理機構】コピー数多型と精神疾患

  仲西 萌絵, 内匠 透

  (公財)金原一郎記念医学医療振興財団, Feb. 2014, 生体の科学, 65(1) (1), 16 - 19, Japanese


• 神経細胞内病態と脳内環境 4.神経細胞におけるRNA障害と脳内環境の関連研究

  黒坂哲, 黒坂哲, 内匠透, 内匠透

  (株)メディカルドゥ, 2014, 遺伝子医学MOOK, (26) (26), 43 - 47, Japanese


• 自閉症の分子基盤 自閉症のモデル動物

  岸本恵子, 野村淳, 内匠透

  (株)先端医学社, 2014, 分子精神医学, 14(2) (2), 119 - 124, Japanese


• 【概日リズムと疾患-病態・診断・治療の最新知見-】概日リズムと疾患 自閉症

  内匠 透

  (株)日本臨床社, Dec. 2013, 日本臨床, 71(12) (12), 2179 - 2181, Japanese


• 自閉症スペクトラム障害治療薬探索 創薬を目指した自閉症ヒト型マウスモデルの開発

  内匠 透

  日本臨床精神神経薬理学会・日本神経精神薬理学会, Oct. 2013, 日本臨床精神神経薬理学会・日本神経精神薬理学会合同年会プログラム・抄録集, 23回・43回, 124 - 124, Japanese


• 概日リズムとうつ病様行動の分子相関 GSK3βによるPER2リン酸化

  山脇 洋輔, 高野 敦子, 畠中 史幸, 玉田 紘太, 仲西 萌絵, 明 智煥, 内匠 透

  日本臨床精神神経薬理学会・日本神経精神薬理学会, Oct. 2013, 日本臨床精神神経薬理学会・日本神経精神薬理学会合同年会プログラム・抄録集, 23回・43回, 234 - 234, Japanese


• 新規時計遺伝子Chronoの統合的解析

  郷力 昭宏, 畠中 史幸, Myung Jihwan, 寄高 崇志, 田ノ上 信太郎, 阿部 高也, 清成 寛, 藤本 勝巳, 加藤 幸夫, 松原 昭郎, 内匠 透

  (公社)日本生化学会, Sep. 2013, 日本生化学会大会プログラム・講演要旨集, 86回, 1T18a - 02, Japanese


• 概日リズムと医学研究

  仲西 萌絵, 内匠 透

  (一社)日本脳循環代謝学会, Jul. 2013, 脳循環代謝, 24(2) (2), 45 - 47, Japanese


• シナプス 構造、機能とその破綻 シナプスの破綻としての発達障害

  岸本 恵子, 内匠 透

  (一社)日本生理学会, Mar. 2013, 日本生理学雑誌, 75(2) (2), 82 - 83, Japanese


• 創薬を目指した精神疾患のヒト型マウスモデル

  内匠 透

  (公社)日本薬学会, Mar. 2013, 日本薬学会年会要旨集, 133年会(1) (1), 31 - 31, Japanese


• ゲノム工学的手法を用いた自閉症モデルマウスの新奇性および低頻度性に対する反応

  岡田 佳奈, 武田 梢, 氏田 麻美, 崎本 裕也, 玉田 紘太, 内匠 透, 坂田 省吾

  日本動物心理学会, Jan. 2013, 動物心理学研究, 62(2) (2), 203 - 203, Japanese


• 【動物モデルから見た発達障害研究】自閉症の病態生理の解明のための染色体15q領域重複マウスモデル(A mouse model of 15q duplication towards understanding of the pathophysiology of autism)

  仲西 萌絵, 内匠 透

  日本生物学的精神医学会, Dec. 2012, 日本生物学的精神医学会誌, 23(4) (4), 267 - 271, Japanese


• 概日リズムと医学研究

  内匠 透

  (一社)日本脳循環代謝学会, Nov. 2012, 脳循環代謝, 24(1) (1), 93 - 93, Japanese


• 精神科領域の用語解説 Duplication of human chromosome 15q11-q13

  内匠 透

  (株)先端医学社, Oct. 2012, 分子精神医学, 12(4) (4), 300 - 302, Japanese


• RNA-binding domain of TLS interacts with nucleoporin

  R. Fujii, T. Takumi

  Oct. 2012, JOURNAL OF NEUROCHEMISTRY, 123, 120 - 120, English

  Summary international conference


• 【心と体のクロストークから解く 精神・神経疾患 発症基盤・病態生理を担う分子カスケードから臨床応用まで】(第I部)精神・神経疾患の最前線 心と体のクロストーク (第1章)精神・神経疾患の最近のトピックス 体との接点 自閉症と睡眠・消化管障害

  内匠 透

  (株)羊土社, Aug. 2012, 実験医学, 30(13) (13), 2018 - 2021, Japanese


• ニューロサイエンスの最新情報 自閉症の動物モデル

  福本 景太, 内匠 透

  (株)中外医学社, Mar. 2012, Clinical Neuroscience, 30(3) (3), 345 - 348, Japanese


• Animal Models of Psychiatric Disorders That Reflect Human Copy Number Variation

  Jun Nomura, Toru Takumi

  2012, NEURAL PLASTICITY, English

  Book review


• 自閉症ヒト型モデルマウスを用いた社会行動の責任神経領域の解析

  内匠 透

  (公財)上原記念生命科学財団, Dec. 2011, 上原記念生命科学財団研究報告集, 25, 1 - 5, Japanese


• 【精神発達遅滞・自閉症の分子医学】発達障害・自閉症のあらたなモデルマウスと分子病態 ヒト型自閉症マウスモデル

  内匠 透

  医歯薬出版(株), Nov. 2011, 医学のあゆみ, 239(6) (6), 708 - 712, Japanese


• 自閉症のモデルとしてのヒト染色体領域重複モデルマウス

  内匠 透

  (株)北隆館, Nov. 2011, BIO Clinica, 26(12) (12), 1146 - 1150, Japanese


• 【モデル動物が拓く新しい精神薬理】自閉症ヒト型モデルマウスの作製と解析

  内匠 透

  (一社)日本神経精神薬理学会, Nov. 2011, 日本神経精神薬理学雑誌, 31(5-6) (5-6), 219 - 222, Japanese


• A mouse model for human chromosome abnormality

  Toru Takumi

  Nov. 2011, Japanese Journal of Neuropsychopharmacology, 31(5-6) (5-6), 219 - 222, Japanese

  Book review


• ヒト型自閉症マウスモデル

  五林 優子, 内匠 透

  (株)医学書院, Oct. 2011, BRAIN and NERVE: 神経研究の進歩, 63(10) (10), 1111 - 1116, Japanese


• A mouse model with human chromosomal abnormality observed in autism

  Yuko Gobayashi, Toru Takumi

  01 Oct. 2011, Brain and Nerve, 63(10) (10), 1111 - 1116, Japanese

  Book review


• 【精神疾患の基礎と臨床】総論 生物学的異常としての精神疾患

  内匠 透

  (株)ニュー・サイエンス社, Sep. 2011, Medical Science Digest, 37(10) (10), 395 - 397, Japanese


• 自閉症ヒト型モデルマウスの開発

  玉田 紘太, 中井 信裕, 内匠 透

  (公社)日本生化学会, Sep. 2011, 生化学, 83(9) (9), 841 - 845, Japanese


• 自閉症責任領域であるヒト染色体15q11-13領域重複マウスの解析

  玉田紘太, 友永省三, 畠中史幸, 中井信裕, 内匠透

  Sep. 2011, 日本神経科学学会, Japanese

  Summary national conference


• 心の発達障害モデルマウスを用いた異常シグナル伝達系の解析

  内匠 透

  (公財)成長科学協会, Aug. 2011, 成長科学協会研究年報, (34) (34), 157 - 159, Japanese


• ヒト染色体15q11-13重複モデルマウスにおける興奮・抑制性神経の形態学的解析

  中井 信裕, 渡辺 康仁, 都甲 めぐみ, 廣木 遥, 三嶋 亮, 笹西 美和子, 高橋 哲也, 松本 昌泰, 玉田 紘太, 内匠 透

  (一社)日本解剖学会, Mar. 2011, 解剖学雑誌, 86(1) (1), 18 - 18, Japanese


• ヒト型マウスモデルによる自閉症の病態解明と新規治療薬開発の基盤研究

  内匠 透, 玉田 紘太, 友永 省三, 畠中 史幸, 中井 信裕

  (公財)先進医薬研究振興財団, Mar. 2011, 精神薬療研究年報, (43) (43), 53 - 54, Japanese


• 身体活動時系列にみる行動組織化とその生成機序精神行動異常の統一的理解に向けて

  中村亨, 菊地裕絵, 吉内一浩, 内匠透, 山本義春

  2011, 生体・生理工学シンポジウム論文集(CD-ROM), 26th


• 脳の科学Up Date 自閉症のモデル動物

  内匠 透

  (株)金芳堂, Jan. 2011, 脳21, 14(1) (1), 78 - 83, Japanese


• Analysis of chromosome engineered mouse model of 15q duplication

  Kota Tamada, Shozo Tomonaga, Fumiyuki Hatanaka, Nobuhiro Nakai, Toru Takumi

  2011, NEUROSCIENCE RESEARCH, 71, E75 - E76, English

  Summary international conference


• Genome-wide profiling of the core clock protein BMAL1 targets reveals strict relationship with metabolism

  Fumiyuki Hatanaka, Chiaki Matsubara, Jihwan Myung, Takashi Yoritaka, Naoko Kamimura, Shuichi Tsutsumi, Akinori Kanai, Yutaka Suzuki, Hiroyuki Aburatani, Sumio Sugano, Toru Takumi

  2011, NEUROSCIENCE RESEARCH, 71, E53 - E53, English

  Summary international conference


• The mouse model of 15q duplication as a developmental brain disorder

  Toru Takumi

  2011, NEUROSCIENCE RESEARCH, 71, E37 - E37, English

  Summary international conference


• Behaviour Organization of Locomotor Activity in Animals and its Mechanism

  NAKAMURA Toru, TAKUMI Toru, YOSHIUCHI Kazuhiro, YAMAMOTO Yoshiharu

  計測自動制御学会, 10 Dec. 2010, 計測と制御 = Journal of the Society of Instrument and Control Engineers, 49(12) (12), 844 - 849, Japanese


• Postnatal development of synaptic function and its disorder 自閉症モデルとしての染色体異常(Postnatal development of synaptic function and its disorder Chromosomal abnormality as a model of autism)

  内匠 透

  (公社)日本生化学会, Dec. 2010, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回, 1S13 - 6, English


• 疾患エピジェネティクスの最前線 インプリンティング領域であるヒト染色体15q11-13相同領域重複マウスの解析

  玉田 紘太, 中谷 仁, 畠中 史幸, 中井 信裕, 大塚 晋, 内匠 透

  (公社)日本生化学会, Dec. 2010, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回, 4W8 - 6, Japanese


• 転写因子BMAL1のゲノム網羅的研究

  畠中 史幸, 松原 千明, 寄高 崇志, 上村 直子, 堤 修一, 金井 昭教, 鈴木 穣, 油谷 浩幸, 菅野 純夫, 内匠 透

  (公社)日本生化学会, Dec. 2010, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回, 2T12 - 10, Japanese


• 染色体工学を用いて作製したヒト染色体15q11-13相同領域重複マウスの行動および脳内モノアミン解析

  玉田 紘太, 友永 省三, 畠中 史幸, 中井 信裕, 高雄 啓三, 宮川 剛, 内匠 透

  (公社)日本生化学会, Dec. 2010, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回, 2T4 - 10, Japanese


• 筋萎縮性側索硬化症の新規原因遺伝子Optineurin(New causative gene of amyotrophic lateral sclerosis: Optineurin)

  丸山 博文, 森野 豊之, 伊東 秀文, 和泉 唯信, 萩原 弘一, 内匠 透, 川上 秀史

  (公社)日本生化学会, Dec. 2010, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回, 4T11 - 1, English


• 自閉症のモデルマウス

  内匠 透

  (株)文光堂, Sep. 2010, Medical Practice, 27(9) (9), 1588 - 1589, Japanese


• てんかんモデル動物の研究から臨床応用へ 染色体15q11-13重複のモデルマウスの作製

  内匠 透

  (一社)日本てんかん学会, Sep. 2010, てんかん研究, 28(2) (2), 229 - 229, Japanese


• モデル動物が拓く新しい精神薬理 自閉症ヒト型モデルマウスの作製と解析

  内匠 透

  日本臨床精神神経薬理学会・日本神経精神薬理学会, Sep. 2010, 日本臨床精神神経薬理学会・日本神経精神薬理学会合同年会プログラム・抄録集, 20回・40回, 68 - 68, Japanese


• 心の発達障害モデルマウスを用いた生化学及び栄養学的解析

  内匠 透

  (公財)成長科学協会, Aug. 2010, 成長科学協会研究年報, (33) (33), 131 - 132, Japanese


• 生体リズム研究の新展開 時計遺伝子のジェネティクス、エピジェネティクス、そして臨床応用まで 時計と気分

  内匠 透

  日本自律神経学会, Aug. 2010, 自律神経, 47(4) (4), 301 - 301, Japanese


• 臨床に必要な神経薬理・化学 概日時計と気分障害

  内匠 透

  (株)中外医学社, Jul. 2010, Clinical Neuroscience, 28(7) (7), 720 - 721, Japanese


• 【拡大・進展を続けるRNA研究の最先端 長鎖noncoding RNA・small RNAからRNA修飾・編集・品質管理まで】noncoding RNAの分子機構と生命現象、疾患との関わり noncoding RNAと精神疾患

  内匠 透

  (株)羊土社, Jun. 2010, 実験医学, 28(10) (10), 1509 - 1514, Japanese


• 発達障害モデルとしてのヒト染色体15q11-13重複モデルマウスの作製

  内匠 透

  (一社)日本解剖学会, Jun. 2010, 解剖学雑誌, 85(2) (2), 80 - 80, Japanese


• 自閉症ヒト型マウスモデルの開発と小児神経学への展開

  内匠 透

  (一社)日本小児神経学会, May 2010, 脳と発達, 42(Suppl.) (Suppl.), S87 - S87, Japanese


• ここまでわかった精神疾患の脳内メカニズム

  橋本 亮太, 森信 繁, 内匠 透, 加藤 忠史

  (公社)日本精神神経学会, May 2010, 精神神経学雑誌, (2010特別) (2010特別), S - 166, Japanese


• 自閉症ヒト型モデルマウスの開発

  内匠 透

  (公社)日本医師会, Mar. 2010, 日本医師会雑誌, 138(12) (12), 2575 - 2577, Japanese


• 【脳神経系の情報伝達と疾患 さまざまなシグナル伝達因子の異常が引き起こす精神疾患・発達障害・神経変性疾患のメカニズム】発達障害 染色体工学を用いた発達障害のヒト型モデルマウスの作製と解析

  内匠 透

  (株)羊土社, Mar. 2010, 実験医学, 28(5) (5), 693 - 698, Japanese


• 染色体工学による自閉症モデルマウスを用いた病態解明及びセロトニン系薬剤の関連研究

  内匠 透, 玉田 紘太, 畠中 史幸, 渡辺 康仁, 中井 信裕, 大塚 晋

  (公財)先進医薬研究振興財団, Mar. 2010, 精神薬療研究年報, (42) (42), 53 - 54, Japanese


• 【脳科学のモデル実験動物】発達障害ヒト型モデルマウス

  内匠 透

  (公財)金原一郎記念医学医療振興財団, Feb. 2010, 生体の科学, 61(1) (1), 65 - 70, Japanese


• 生体リズム研究の新展開 時計遺伝子のジェネティクス、エピジェネティクス、そして臨床応用まで 時計と気分

  内匠 透

  日本自律神経学会, Nov. 2009, 日本自律神経学会総会プログラム・抄録集, 62回, 76 - 76, Japanese


• 行動をもたらす分子の生化学 概日リズムと気分障害(Molecular correlations between circadian rhythm and mood disorder)

  内匠 透

  (公社)日本生化学会, Sep. 2009, 日本生化学会大会プログラム・講演要旨集, 82回, 1S7a - 4, English


• うつ病ラットモデルを用いたリチウムシグナルとサーカディアンリズム解析

  高野 敦子, 内匠 透

  (公社)日本生化学会, Sep. 2009, 日本生化学会大会プログラム・講演要旨集, 82回, 4T9a - 3, Japanese


• 【発達障害の新たな遺伝子】自閉症モデルとしてのヒト染色体15q11-13重複モデル

  内匠 透

  (株)先端医学社, Jul. 2009, 分子精神医学, 9(3) (3), 234 - 239, Japanese


• Thanks to Anatomists

  TAKUMI T.

  01 Mar. 2009, 解剖學雜誌, 84(1) (1), 29 - 29, Japanese


• システム生物学による機能解剖学 視交叉上核の機能解剖学

  明 智煥, 畠中 史幸, 中村 渉, 内匠 透

  (一社)日本解剖学会, Mar. 2009, 解剖学雑誌, 84(Suppl.) (Suppl.), 83 - 83, English


• ミオシンVによるRNA輸送の制御機構の解明

  内匠 透

  (公財)ライフサイエンス振興財団, Mar. 2009, ライフサイエンス振興財団年報, 23, 38 - 40, Japanese


• A humanoid mouse model for autism by a chromosome engineering

  Toru Takumi

  2009, NEUROSCIENCE RESEARCH, 65, S27 - S27, English

  Summary international conference


• The chromosome-engineered mouse model for human chromosome 15q11-13 duplication

  Toru Takumi

  2009, JOURNAL OF PHARMACOLOGICAL SCIENCES, 109, 29P - 29P, English

  Summary international conference


• ヒト由来BMALI:CLOCKのbHLHドメインの精製および結晶化

  坂井恵理子, 内匠透, 加藤博章, 中津亨

  2009, 日本薬学会年会要旨集,129th,4,129


• 精神疾患の分子統合的アプローチ 染色体工学的手法による自閉症モデルマウス(A molecular approach towards an understanding of mind and behavior)

  内匠 透

  (公社)日本生化学会, Nov. 2008, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 81回・31回, 4S25 - 5, English


• Wnt5a/Ror2経路はMMP-13を介して骨肉腫細胞のinvadopodia形成と浸潤を制御する

  榎本 真宏, 早川 沙織, 任 大勇, 玉田 紘太, 内匠 透, 西田 満, 南 康博

  (公社)日本生化学会, Nov. 2008, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 81回・31回, 3T12 - 6, Japanese


• 【神経回路の制御と脳機能発現のメカニズム 細胞移動・接着から記憶・学習・行動発現までの分子基盤解明と研究を躍進させる最新技術】神経回路の破綻による疾患 自閉症モデルマウス

  内匠 透

  (株)羊土社, Jul. 2008, 実験医学, 26(12) (12), 2009 - 2014, Japanese


• 小児神経疾患とエピジェネティクス 染色体工学的手法によるヒト染色体15q11-13重複モデルマウスの作製

  内匠 透

  (一社)日本小児神経学会, May 2008, 脳と発達, 40(Suppl.) (Suppl.), S132 - S132, Japanese


• Bmal1 KOマウスの給餌前行動

  中村渉, 中村渉, 畠中史幸, 畠中史幸, 内匠透, 内匠透

  2008, 時間生物学, 14(2) (2)


• デュアルカラールシフェラーゼマウスによるBmal1とPer2の同時発現解析

  野口貴子, 道畑朋子, 中村渉, 内匠透, 池田正明, 池田正明, 近江谷克裕, 近江谷克裕, 中島芳浩

  2008, 時間生物学, 14(2) (2)


• ヒトとマウスにおける行動普遍則とその崩壊

  中村亨, 内匠透, 高野敦子, 青柳直子, 吉内一浩, ZBIGNIEW Struzik, 山本義春

  2008, 日本生体医工学会大会プログラム・論文集(CD-ROM), 47th


• 時計遺伝子による細胞時間制御の分子機構

  内匠 透

  (公財)上原記念生命科学財団, Dec. 2007, 上原記念生命科学財団研究報告集, 21, 156 - 158, Japanese


• 【RNA研究の新展開】精神機能とRNA

  内匠 透

  (株)ニュー・サイエンス社, Nov. 2007, 細胞, 39(12) (12), 505 - 508, Japanese


• 嗅覚発達におけるFezf1の役割(Fezf1 is important for penetration of CNS basal lamina by olfactory sensory axons)

  渡辺 康仁, 井上 浄, 山本 綾子, 中谷 仁, 丹生 健一, 寺島 俊雄, 近藤 玄, 竹田 潤二, 内匠 透

  (公社)日本生化学会, Nov. 2007, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 80回・30回, 4T14 - 10, English


• 【中枢時計と末梢時計 時計遺伝子研究の最前線】疾患との関連 リチウムと概日時計とうつ病

  内匠 透

  (株)中外医学社, Oct. 2007, Clinical Neuroscience, 25(10) (10), 1156 - 1157, Japanese


• 心臓性急死は時間医学の手法で予測できるか"Chronomics and sudden cardiac death" 時計遺伝子の転写振動及びその分子メカニズム

  内匠 透

  日本自律神経学会, Aug. 2007, 自律神経, 44(4) (4), 246 - 250, Japanese


• 精神疾患における器質的障害部位としてのスパインバイオロジー

  内匠 透

  (一社)日本神経精神薬理学会, Jun. 2007, 日本神経精神薬理学雑誌, 27(3) (3), 103 - 107, Japanese


• 精神行動異常の分子戦略

  内匠 透, 堀尾 嘉幸

  札幌医科大学, Jun. 2007, 札幌医学雑誌, 76(1-3) (1-3), 21 - 21, Japanese


• Spine biology in psychiatric diseases

  Toru Takumi

  Jun. 2007, Japanese Journal of Neuropsychopharmacology, 27(3) (3), 103 - 107, Japanese

  Book review


• 線維芽細胞を用いた抗うつ薬スクリーニング系の確立

  内匠 透, 高野 敦子, 榊田 容子, 松原 千明, 須藤 智美, 石橋 拓也, 仙波 純一, 渡辺 明子, 吉川 武男

  (公財)先進医薬研究振興財団, Mar. 2007, 精神薬療研究年報, (39) (39), 232 - 235, Japanese


• Is RNA involved in autistic behaviors?

  Toru Takumi

  2007, NEUROSCIENCE RESEARCH, 58, S12 - S12, English

  Summary international conference


• 精神疾患はスパインの病気?

  内匠 透

  共立出版(株), Dec. 2006, 蛋白質・核酸・酵素, 51(15) (15), 2328 - 2333, Japanese


• 心臓性急死は時間医学の手法で予測できるか 時計遺伝子の転写振動及びその分子メカニズム

  内匠 透

  日本自律神経学会, Nov. 2006, 日本自律神経学会総会プログラム・抄録集, 59回, 55 - 55, Japanese


• 遺伝子改変マウスを用いた行動科学 ゲノム工学を用いたヒト精神行動異常モデルマウス

  内匠 透

  日本神経化学会, Aug. 2006, 神経化学, 45(2-3) (2-3), 267 - 267, Japanese


• Dendritic spine morphology and local translation of actin-related proteins

  R. Fujii, T. Takumi

  Jul. 2006, JOURNAL OF NEUROCHEMISTRY, 98, 48 - 48, English

  Summary international conference


• 分子時計の概日転写制御機構

  内匠 透

  (公社)日本生化学会, May 2006, 生化学, 78(5) (5), 425 - 429, Japanese


• リチウムシグナルとサーカディアンリズムからみた感情障害新規創薬標的分子の同定解析

  内匠 透, 高野 敦子, 島 悠香理, 榊田 容子, 須藤 智美, 松原 千明, 仙波 純一, 渡辺 明子, 吉川 武男

  (公財)先進医薬研究振興財団, Mar. 2006, 精神薬療研究年報, (38) (38), 237 - 241, Japanese


• からくり分子時計

  内匠 透

  (株)学研メディカル秀潤社, Jan. 2006, 細胞工学, 25(2) (2), 174 - 178, Japanese


• Fez2欠損マウスの嗅球細胞構築異常と嗅神経投射異常

  奥山 綾子[山本], 渡辺 康仁, 井上 浄, 内匠 透, 寺島 俊雄

  (一社)日本解剖学会, Mar. 2005, 解剖学雑誌, 80(Suppl.) (Suppl.), 217 - 217, Japanese


• 抗精神病薬標的分子としてのRNA結合蛋白TLSの分子機能解析

  内匠 透, 藤井 律子, 吉村 淳, 渡辺 康仁, 田中 正視, 玉田 鉱太, 榊田 容子, 島 悠香里, 須藤 智美, 岡部 繁男, 西川 徹

  (公財)先進医薬研究振興財団, Mar. 2005, 精神薬療研究年報, (37) (37), 151 - 154, Japanese


• マウス概日リズム制御機構におけるId2の役割

  足立明人, 長野護, 升本宏平, 藤岡厚子, 内匠透, 横田義史, 重吉康史

  2005, 時間生物学, 11(2) (2)


• 大脳新皮質特異的遺伝子の検索 皮質第5,6層特異的遺伝子Fezの同定

  井上 浄, 寺島 俊雄, 西川 徹, 内匠 透

  (一社)日本解剖学会, Sep. 2004, 解剖学雑誌, 79(3) (3), 109 - 109, Japanese


• The receptor tyrosine kinase Ror2 associates with and is activated by casein kinase I epsilon

  Shuichi Kani, Isao Oishi, Hiroyuki Yamamoto, Akinori Yoda, Hiroaki Suzuki, Akira Nomachi, Akira Kikuchi, Toru Takumi, Yasuhiro Minami

  May 2004, CELL STRUCTURE AND FUNCTION, 29, 76 - 76, English

  Summary international conference


• 抗精神病薬標的分子としてのRNA結合蛋白の機能解析

  内匠 透, 藤井 律子, 吉村 淳, 井上 浄, 渡辺 康仁, 漆戸 智恵, 岡部 繁男, 西川 徹

  (公財)先進医薬研究振興財団, Mar. 2004, 精神薬療研究年報, (36) (36), 90 - 94, Japanese


• ゲノムスクリーニングにより得られた抗精神病薬標的分子の機能解析

  内匠 透, 井上 浄, 藤井 律子, 中谷 仁, 渡辺 康仁, 吉村 淳, 松原 千明, 榊田 容子, 濱島 久美子, 山田 敦子, 油谷 浩幸, 海野 麻未, 西川 徹

  (公財)先進医薬研究振興財団, Mar. 2003, 精神薬療研究年報, (35) (35), 149 - 153, Japanese


• 生活習慣病における医学,薬学の萠芽的研究 生活習慣病の基盤としての生体リズムの分子機構解明

  内匠 透

  (公財)鈴木謙三記念医科学応用研究財団, Jan. 2001, 医科学応用研究財団研究報告, 18, 121 - 124, Japanese


• 薬物動態ならびに薬物に対する感受性に影響を及ぼす生体リズムの機構解明

  内匠 透, 岡村 均

  (公財)臨床薬理研究振興財団, May 2000, 臨床薬理の進歩, (21) (21), 79 - 83, Japanese


• 【新遺伝子工学ハンドブック】遺伝子クローニング 発現クローニング アフリカツメガエル卵母細胞を用いた発現クローニング

  内匠 透

  (株)羊土社, Sep. 1999, 実験医学, 別冊(新遺伝子工学ハンドブック) (新遺伝子工学ハンドブック), 28 - 35, Japanese


• 哺乳類Timelessの分子解析

  内匠 透, 永峰 恭子, 三宅 茂, 松原 千明, 田口 浩司, 武木田 誠一, 榊田 容子, 西川 和子, 岸本 利彦, 丹羽 真一郎, 岡村 均

  (公社)日本生化学会, Aug. 1999, 生化学, 71(8) (8), 999 - 999, Japanese


• 成長ホルモン分泌における日周リズムの分子機構の解明

  内匠 透, 岡村 均

  (公財)成長科学協会, Jul. 1999, 成長科学協会研究年報, (22) (22), 289 - 300, Japanese


• 【細胞のシグナリング】 哺乳類時計遺伝子mPer

  岡村 均, 重吉 康史, 内匠 透, 山口 瞬, 八木田 和弘

  (株)中山書店, May 1999, Molecular Medicine, 36(臨増 細胞シグナルリング) (臨増 細胞シグナルリング), 315 - 324, Japanese


• 光依存性位相変位と哺乳類ピリオド遺伝子

  重吉 康史, 田口 浩司, 山本 秀三, 内匠 透, 岡村 均

  (一社)日本解剖学会, Apr. 1999, 解剖学雑誌, 74(2) (2), 255 - 255, Japanese


• サーカディアンリズムの分子レベルでの解析 時計遺伝子の同定と老化

  岡村 均, 内匠 透, 重吉 康史, 閻 莉莉

  (一社)兵庫県医師会, Mar. 1999, 兵庫県医師会医学雑誌, 41(3) (3), 102 - 103, Japanese


• 【転写因子・研究1999】神経形成と神経機能にかかわる転写因子 転写因子による生物時計の制御

  内匠 透, 岡村 均

  (株)羊土社, Feb. 1999, 実験医学, 17(3) (3), 372 - 378, Japanese


• マウスper遺伝子導入ショウジョウバエの解析

  重吉 康史, 八木田 和弘, 内匠 透, アミタ・セーガル, 岡村 均

  (一社)日本解剖学会, Feb. 1999, 解剖学雑誌, 74(1) (1), 40 - 40, Japanese


• mPer3cDNAのクローニングとゲノム解析

  高嶋 直敬, 内匠 透, 重吉 康史, 岡村 均

  (一社)日本解剖学会, Feb. 1999, 解剖学雑誌, 74(1) (1), 78 - 78, Japanese


• リズム性及び居眠り事故予防のための生体リズムの機構解明

  内匠 透, 岡村 均

  佐川交通社会財団, 1999, 交通安全対策振興助成研究報告書 一般研究, (14) (14), 100 - 104, Japanese


• 脳を知る 体内時計とその遺伝子

  内匠 透, 岡村 均

  (株)学研メディカル秀潤社, Dec. 1998, 細胞工学, 17(12) (12), 1954 - 1962, Japanese


• 光不応性時計遺伝子マウスmPer3の単離とサーカデイアンリズム

  内匠透, 田口浩司, 三宅茂, 榊田容子, 高嶋直敬, 松原千明, 前林佳朗, 岡村均

  Nov. 1998, 日本分子生物学会年会プログラム・講演要旨集, 21st, 657, Japanese


• ほ乳類時計遺伝子mPer2の単離と発現解析

  内匠透, 松原千明, 重吉康史, 田口浩司, 八木田和弘, 前林佳朗, 榊田容子, 高嶋直敬, 岡村均

  Nov. 1998, 日本分子生物学会年会プログラム・講演要旨集, 21st, 657, Japanese


• ラット大脳皮質､小脳皮質におけるrPer1, rPer2 mRNAの日内リズム

  武木田 誠一, 閻 莉莉, 内匠 透, 岡村 均

  01 Oct. 1998, 日本時間生物学会会誌: Journal of Chronobiology, 4(2) (2), 109 - 109, Japanese


• ラット視交叉上核におけるper1,per2 mRNAの日内リズムと光による誘導

  閻 莉莉, 武木田 誠一, 内匠 透, 岡村 均

  日本神経化学会, Sep. 1998, 神経化学, 37(3) (3), 533 - 533, Japanese


• サーカディアンクロックの分子機構 哺乳類Periodの分子多様性

  内匠 透, 岡村 均

  (公社)日本生化学会, Aug. 1998, 生化学, 70(8) (8), 683 - 683, Japanese


• 成長ホルモン分泌における日周リズムの分子機構の解析

  内匠 透, 岡村 均

  (公財)成長科学協会, Jul. 1998, 成長科学協会研究年報, (21) (21), 375 - 387, Japanese


• 哺乳類時計遺伝子

  岡村 均, 内匠 透, 重吉 康史

  (株)羊土社, Jun. 1998, 実験医学, 16(9) (9), 1193 - 1197, Japanese


• 発生特異的 differential mRNA display 法による視交叉上核に発現する新規転写因子のクロｰニング

  前林 佳朗, 重吉 康史, 中村 亨, 山本 秀三, 内匠 透, 岡村 均

  01 Oct. 1997, 日本時間生物学会会誌: Journal of Chronobiology, 3(2) (2), 67 - 67, Japanese


• マウス脳における period mRNA と clock mRNA の発現

  重吉 康史, 山本 秀三, 田口 浩司, 武木田 誠一, 前林 佳朗, 高嶋 直敬, 程 肇, 榊 佳之, 内匠 透, 岡村 均

  01 Oct. 1997, 日本時間生物学会会誌: Journal of Chronobiology, 3(2) (2), 66 - 66, Japanese


• ATP依存性内向き整流カリウムチャネルROMK1(Kir1.1)のisoformのクローニングおよびその機能と発現分布

  近藤 千香子, 磯本 正二郎, 松本 重人, 高橋 尚彦, 内匠 透, 山下 静也, 竹村 芳, 松沢 佑次, 倉智 嘉久

  01 Aug. 1996, 日本分子生物学会年会プログラム・講演要旨集, 19, 312 - 312, Japanese


• 内向き整流カリウムチャネルK_-2(Kir4.1)の星状膠細胞における発現の検討

  内匠 透, 堀尾 嘉幸, 近藤 千賀子, 高橋 尚彦, 小山 豊, 馬場 明道, 稲野辺 厚, 伊藤 稔, 倉智 嘉久

  01 Aug. 1996, 日本分子生物学会年会プログラム・講演要旨集, 19, 312 - 312, Japanese


• 内向き整流カリウムチャネルKAB-2(Kir4.1)の星状膠細胞における発現の検討

  内匠 透

  (公社)日本生化学会, Jul. 1996, 生化学, 68(7) (7), 824 - 824, Japanese


• 遺伝子クローニング cDNAクローニング Xenopus oocytesを用いた発現クローニング

  内匠 透

  (株)羊土社, Apr. 1996, 実験医学, 別冊(新遺伝子工学ハンドブック) (新遺伝子工学ハンドブック), 50 - 57, Japanese


• グリア細胞特異的内向き整流性K+チャネル

  内匠 透

  (公社)日本生化学会, Jul. 1995, 生化学, 67(7) (7), 955 - 955, Japanese


• MOLECULAR CHARACTERIZATION OF A NOVEL SUBFAMILY OF INWARD RECTIFIER POTASSIUM CHANNELS

  T TAKUMI, T ISHI, K SOHMIYA, KI MORISHIGE, N TAKAHASHI, M YAMADA, S NAKANISHI, Y KURACHI

  Dec. 1994, JOURNAL OF GENERAL PHYSIOLOGY, 104(6) (6), A9 - A9, English

  Summary international conference


• MOLECULAR-BASIS OF ISK PROTEIN-REGULATION BY OXIDATION AND CHELATION

  AE BUSCH, S WALDEGGER, T HERZER, G RABER, E GULBINS, R SWANSON, K FOLANDER, T TAKUMI, K MORIYOSHI, S NAKANISHI, F LANG

  Sep. 1994, JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY, 5(3) (3), 283 - 283, English

  Summary international conference


• 腎と分子生物学 Small K+ channel (Isk) その後

  内匠 透

  (株)東京医学社, Sep. 1992, 腎と透析, 33(3) (3), 513 - 517, Japanese


• K+チャンネル活性を示す新しい膜蛋白の分子解析

  内匠 透, 大久保 博晶, 中西 重忠

  (株)東京医学社, Sep. 1991, 腎と透析, 31(3) (3), 651 - 654, Japanese


• K+イオンチャンネル活性を有する新しい膜蛋白(Isk)の単離と構造,機能解析

  内匠 透

  (公社)日本生化学会, Sep. 1989, 生化学, 61(9) (9), 1193 - 1193, Japanese

• Social circuits and its dysfunction in autism spectrum disorders

  内匠 透, 中井 信裕, 棒田 亜耶花

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Fund for the Promotion of Joint International Research (International Collaborative Research), Kobe University, 08 Sep. 2023 - 31 Mar. 2026


• 自閉症ヒト脳オルガノイドの表現解析による病態の基礎的理解

  内匠 透

  日本学術振興会, 科学研究費助成事業 基盤研究(A), 基盤研究(A), 神戸大学, 05 Apr. 2021 - 31 Mar. 2024

  自閉症は社会性の障害に代表される発達障害であり、その発症にはコピー数多型（CNV）等の遺伝的寄与が示唆されている。我々は世界に先駆けてCNVの自閉症ヒト型モデルマウス、また自閉症細胞モデル（Autism in a dish）として、独自に開発した次世代染色体工学を用いて網羅的マウス胚性幹（ES）細胞ライブラリーを構築した。本研究では、ヒトES細胞モデルを用いて、神経をはじめとする２次元培養並びに３次元オルガノイド脳培養を組み合わせて、分子、形態から機能に至る多面的な解析を取り入れることにより、ヒト病態の総合的理解を得る。本成果は、自閉症を含む精神疾患の病態パスウェイやハブ遺伝子などの創薬シーズに繋がるだけでなく、CNVの発現制御機構の解明というゲノム異常の基盤的理解をもたらす。


• レジリエンスの新基礎医学的理解への挑戦

  内匠 透

  日本学術振興会, 科学研究費助成事業 挑戦的研究(萌芽), 挑戦的研究(萌芽), 神戸大学, 09 Jul. 2021 - 31 Mar. 2023

  本研究では、概日リズムとうつの関連研究から偶発的に生まれたレジリエンスモデルをもとに、レジリエンスの分子的基盤である時計タンパク質のキナーゼを同定し、その阻害剤の開発とともに、これまでにない全く新しい作用機序の気分安定薬、抗うつ薬の開発の基盤を提供する。またレジリエンスの分子カスケードを明らかにする。さらにレジリエンスの標的脳部位を探索する事を目的とする。 時計タンパク質PER2の変異体ノックイン（KI）マウスをレジリエンスモデルとして確立した。変異体の周辺アミノ酸配列はキナーゼであるGSK3betaのリン酸化モチーフ「S-X-X-X-S-P」に対応しており、GSK3betaによるN末端側のセリンのリン酸化にはC末端側のセリンがあらかじめ別のプライミングキナーゼによってリン酸化される必要がある。またこの部位はプロリンと隣接しているため、このプライミングキナーゼはプロリン指向性キナーゼである。本研究ではプライミングキナーゼの探索とともに、レジリエンスの分子カスケードを明らかにするためにシグナル分子の網羅的探索、さらにレジリエンスの神経回路を明らかにするために標的脳部位の探索を行う。


• 精神疾患のヒトES細胞モデルを用いたマルチスケール解析

  内匠 透

  日本学術振興会, 科学研究費助成事業 新学術領域研究(研究領域提案型), 新学術領域研究(研究領域提案型), 神戸大学, 01 Apr. 2021 - 31 Mar. 2023

  コピー数多型（CNV）は遺伝的浸透度が高く、病態理解のためのモデルとして適している。CNVとしてヒト染色体1q21.1領域に焦点をあてる。1q21.1欠失・重複は、共通症状として、てんかんの他、それぞれ小頭症・統合失調症、大頭症・自閉症と遺伝子発現依存的に異なる表現型を示すことが知られている。本ヒトES細胞・オルガノイドモデルの形態、機能、オミックス等の多面的な解析により遺伝子発現依存的な病態を明らかにする。また精神疾患のヒト細胞モデルとしては昨今iPS細胞が頻繁に用いられる一方、ES細胞モデルはisogenic変異であるという利点を有しており、マルチスケール解析による成果は、種々の網羅的データだけでなく、iPS細胞研究との補完性を提供する。


• Analysis of gastorintestinal problems in autism mouse models

  内匠 透

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for JSPS Fellows, Kobe University, 13 Nov. 2020 - 31 Mar. 2023

  Autism is a highly prevalent neurological disorder characterized by impaired social interactions and stereotyped or repetitive behaviours. Autism-associated GI symptoms frequently present from early childhood to adulthood and patients with autism are approximately four-fold more likely to be hospitalized with GI disorders. GI function is regulated by the intrinsic enteric nervous system (ENS), a complex network of neurons extending along the length of the GI tract arranged in two plexuses, the submucosal and myenteric plexuses that predominantly regulate gut secretion and motility, respectively. We will characterize the GI phenotype in two preclinical models of autism (CHD8 and 15q mice) in order to identify therapeutic targets to treat GI dysfunction.


• Dynamic regulation of neural circuit remodeling by scrap & build system

  Emoto Kazuo

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), The University of Tokyo, 30 Jun. 2016 - 31 Mar. 2021

  This research group aims to establish a novel concept and technology to figure out the dynamic function of neural circuits. To this end, we have supported the group activities by (1) HP managements (2) organization of annual group meetings (3) sharing research information within the group (4) financial supports for young researchers (5) organization of international meetings.


• Scrap& build of synapse towards understanding neuropsychiatric disorders

  Takumi Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), 30 Jun. 2016 - 31 Mar. 2021

  Takumi found Neuroligin 1 mutation related to synapse formation from genetic analysis of autistic families. The model mouse with the point mutation was produced, and the social behavior was abnormal. An integrated analysis of 15q dup mice as an autism model mouse showed abnormalities in serotonin neurons mainly in the raphe nucleus, E/I imbalance in the somatosensory cortex. These abnormalities were improved by serotonin replacement therapy during the developmental stage. When fMRI under mouse arousal was constructed and 15q dup mice were analyzed, the neuronal functional connectivity was reduced. Our collaborator, Mandai, elucidates the mechanisms by which afadin regulates synaptic formation and synaptic transmission. αN-catenin, a membrane-lining protein of cadherin, NGL -3, an adhesion molecule, and MAGUIN, a molecule in the postsynaptic density, were involved in these mechanisms.


• Biologically integrative research of autism

  Takumi Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (S), Grant-in-Aid for Scientific Research (S), 31 May 2016 - 31 Mar. 2021

  Autism is a developmental disorder with impaired social skills. It is a multifactorial complex disorder with a high genetic contribution, but the involvement of environmental factors cannot be denied, and the elucidation of its pathophysiology is still incomplete. In this study, we clarified one aspect of autism pathology by conducting multidisciplinary analysis at the cellular/synaptic (development of autistic cell models), circuit/behavioral (clarification of neural circuits for abnormal social behavior), and environmental factors (disorders of brain development due to brain-gut interactions) levels, incorporating methodologies that employ the most advanced technologies for each.


• 炎症と腸内フローラに基づく自閉症モデルの病態生理解析

  内匠 透, SAITO VIVIANE

  日本学術振興会, 科学研究費助成事業 特別研究員奨励費, 特別研究員奨励費, 国立研究開発法人理化学研究所, 26 Apr. 2017 - 31 Mar. 2019

  自閉スペクトラム症(ASD)は、社会性の異常を中心とする脳の発達障害である。我々は、ASDモデルマウスであるBTBRマウスが、抗生物質処理にかかわらず野生型B6系統よりも多くの発声を発し、異常なコミュニケーションパターンを示すことを明らかにした。ネオマイシン処理は雄B6およびBTBRマウスにおいてのみUSV数を増加させたが、ミノサイクリンはいずれの群においても総発声数に影響を及ぼさなかった。雌のUSV総数に有意な抗生物質効果は認められなかった。しかし、雌のUSVタイプの詳細な分析では、ミノサイクリンはB6マウスのNSタイプの割合を減少させ、MJタイプを増加させた。これらの結果は、周産期の抗生物質投与が、系統と性別に依存して、タイプと数に影響することを示している。 細菌叢組成に対する抗生物質投与の効果に関しては、ネオマイシンはミノサイクリンより多くの細菌種/属を減少させることにより大きな影響を及ぼすと考えられた。両抗生物質はビフィズス菌を減少させる。ビフィズス菌は、腸粘膜バリアの改善および腸内リポ多糖類を含む一連の有益な健康効果を発揮する可能性がある哺乳類腸微生物叢の主要なプロバイオティクス成分の一つである。ネオマイシンは微生物相および仔のUSVに大きな影響を及ぼした。これらの結果は、脳腸相関の根底にあるシグナル伝達機構を考える上で興味ある。 本研究では、周産期に投与された2種類の異なる抗生物質の母親の微生物相の組成に及ぼす影響と仔の発声に対するそれぞれの影響を評価した。我々の結果は、抗生物質投与が系統と性別に依存してUSVコールの種類と数に影響することを示した。この行動への影響は脳内の活性化ミクログリアの特異的変化を伴っており、両方の過程は、周産期の間の母親の腸内フローラ障害によって引き起こされる異常な神経炎症性プロフィールと相関している可能性がある。


• Analysis of molecular mechanisms of Wnt5a-Ror signaling in repair after tissue damage, inflammation, and cancer progression

  Minami Yasuhiro

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B), Grant-in-Aid for Scientific Research (B), Kobe University, 01 Apr. 2016 - 31 Mar. 2019

  It has been well established that Ror1 and Ror2 receptor tyrosine kinases play important roles in developmental morphogenesis and tissue-/organo-genesis by acting as receptors for Wnt5a. In this study, we have shown that Ror1 and Ror2 play important roles in regulating repair and inflammatory responses after damage of brain neocortex and skeletal muscle by using experimental animal models. Furthermore, we show that Ror2 as well as Ror1 are expressed at high levels in various types of cancer cells, including osteosarcoma cells, breast cancer cells, malignant pleural mesotheliomas, and colo-rectal cancer cells, thereby promoting invasion of these cancer cells by activating similar and/or different signaling pathways.


• 自閉症モデルの病態生理解析

  内匠 透, ZUKO AMILA

  日本学術振興会, 科学研究費助成事業 特別研究員奨励費, 特別研究員奨励費, 国立研究開発法人理化学研究所, 22 Apr. 2016 - 31 Mar. 2018

  自閉症は1)社会的コミュニケーションや社会的相互作用の障害、２）行動、興味や活動の限局、繰り返しパターン、と定義される小児の精神疾患である。自閉症の遺伝的寄与率は他の精神疾患より高いため、特にヒト遺伝学からの研究が進み、遺伝的異常としては、脆弱性X症候群に代表される症候群性のもの、レア変異遺伝子、そしてコピー数多型（copy number variants, CNV）が知られている。 我々の自閉症細胞モデルプロジェクトは、CRISPR/Cas（ゲノム編集技術）を用いた次世代染色体工学を利用して、自閉症のすべてのCNVのES(多能性幹)細胞モデルを構築しようとするものある。細胞モデルの表現型解析として、in vitro で分化した神経細胞を用いて形態・機能解析を行う。また、分化させた神経細胞を脳内に導入し、in vivoでのネットワーク解析に発展させるという計画も含まれる。 マウスES細胞のin vitro神経分化系を確立した。本分化系では、種々の神経細胞だけでなくグリア細胞への分化も可能であることを確認している。 細胞モデルに関して、CGH (comparative genomic hybridization)によるESの評価を行った。複数のCNVモデルのES細胞を神経細胞に分化後、神経突起などの形態学的解析およびKClやグルタミン酸に対するCa反応を見たところ、コントロールベクターを導入した神経細胞との差異を見出した。 一方、in vivoでのネットワーク解析のためのトランスシナプス標識に関しては、ウイルス構成に必須のG膜タンパク質を除いてmCherryを導入した狂犬病ウイルス変異体と感染に必要なEnvA及びその受容体TVA等、トランスシナプス標識に必要なコンストラクト、ウイルスを作製した。また、実際に脳内にコントロールを接種するのに成功した。


• The gut-brain axis examined by wireless optogenetics

  Takumi Toru, YAMANAKA Akihiro

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research, Grant-in-Aid for Challenging Exploratory Research, Institute of Physical and Chemical Research, 01 Apr. 2016 - 31 Mar. 2018

  The gut is called as the second brain and the gut-brain axis is an interesting network between organs for higher brain functions including cognitive function. To understand the higher brain function regulated by the autonomic nervous system, we develop the wire-less optogenetics to physiologically see the activity of the autonomic nervous system and the enteric nerves with relatively lower invasiveness. The results of this study will be a tool to examine the regulatory network between different organs.


• The target gene of Ube3a is implicated in transcriptional regulation

  Furumai Ryohei, TAKUMI Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Institute of Physical and Chemical Research, 21 Oct. 2015 - 31 Mar. 2018

  UBE3A is a responsible gene for the pathogenesis of Angelman syndrome (AS), a neurodevelopmental disorder. Since UBE3A encodes an E3 ubiquitin ligase, several targets have been identified including synaptic molecules. Although proteolysis occurs mainly in the cytoplasm, UBE3A localizes not only in the cytoplasm but also in the nucleus. In fact, UBE3A has been classically known as a transcriptional regulator of nuclear receptor family. However, the function of UBE3A in the nucleus remains unclear especially in neurophysiology. Thus, we focused on the involvement of UBE3A in transcription in the nuclei of neurons. Genome-wide transcriptome analysis revealed that downstream of Interferon Regulatory Factor (IRF) was changed in UBE3A-deficient AS model mice. In vitro biochemical analyses further demonstrated that UBE3A could enhance IRF-dependent transcription. These results suggested the novel function of UBE3A as a transcriptional regulator of immune system in brain.


• Elucidating pathomechanism of ALS through the abnormality in astrocytes

  YAMANAKA Koji, Suzumura Akio, Takeuchi Hideyuki, Takahashi Ryosuke, Yamashita Hirofumi, Endo Fumito, Watanabe Seiji

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B), Grant-in-Aid for Scientific Research (B), Nagoya University, 01 Apr. 2014 - 31 Mar. 2018

  Using model mice for amyotrophic lateral sclerosis (ALS), we identified cytokine TGF-β1 as a detrimental factor for ALS by interfering a neuroprotective environment mediated by glial cells. TGF-β1 was abnormally upregulated in astrocytes of human ALS and a mouse model for ALS. Administrating TGF-β1 inhibitor slowed disease progression in ALS mice, indicating TGF-β1 as a potential therapeutic target for ALS. In addition, we elucidated a novel role of innate immune adaptor TRIF in eliminating abnormally activated astrocytes through inducing apoptosis.


• 自閉症細胞モデルによる病態理解のための表現解析

  内匠 透

  日本学術振興会, 科学研究費助成事業 基盤研究(A), 基盤研究(A), 国立研究開発法人理化学研究所, 01 Apr. 2016 - 31 Mar. 2017

  自閉症は社会性の障害に代表される発達障害であり、その発症にはコピー数多型（CNV）等の遺伝的寄与が示唆されている。我々はCNVの自閉症モデルマウスを開発した経験から、ヒトで報告されている全てのCNVの網羅的胚性幹（ES）細胞ライブラリーを、独自に開発した次世代染色体工学を用いて構築し、シナプスレベルを含むin vitro, in vivoの多面的表現解析を行う事を考案した。 １.自閉症データベースの構築：これまでのバイオインフォマティクス解析で、自閉症のゲノム解析で得られた論文、Web情報からコピー数多型（CNV）を有する自閉症例の網羅的データベースを構築した。 ２.次世代染色体工学によるマウスES細胞モデルライブラリーの構築：我々はTALEN(Transcription activator-like effector nuclease)及びCRISPR/Cas9(Clustered regulatory interspaced short palindromic repeats/CRISPR associated protein)といったゲノム編集技術を組み合わせて、新たに「次世代」染色体工学的手法を既に確立している。ライブラリー構築に用いる次世代染色体工学は、２組のCRISPRベクターと選択マーカーを組み込んだターゲティングベクターを組み合わせる事により、CRISPRのoff-targetの可能性をなくし、かつ薬剤選択による細胞スクリーニングを可能にする。本法を用いて細胞モデルライブラリーの構築を行っている。 ３.細胞分化系の構築：ES細胞ライブラリーのスクリーニングのために、神経細胞（ニューロン）への分化系を構築する。神経幹細胞及び神経細胞への分化系に関しては、既に応募者の研究室で確立した。


• ミトコンドリアＤＮＡがシナプス発達に関与するしくみ

  内匠 透, LIN CHIA WEN

  日本学術振興会, 科学研究費助成事業 特別研究員奨励費, 特別研究員奨励費, 国立研究開発法人理化学研究所, 24 Apr. 2015 - 31 Mar. 2017

  シナプス発達にミトコンドリアDNAが関与するという仮説は、自閉症を含む精神神経疾患の病態の中で炎症との関連が重要視されているという最近の研究状況から来ている。自閉症では、社会性相互作用やコミュニケーションの異常、限局・反復する興味や行動といった社会性の異常を中心とした主症状を示すが、その他に胃腸障害は頻繁に見られる随伴症状である。 我々はミトコンドリアDNAの異常そのものは見いだすことができなかったが、自閉症モデルマウスの脳内での炎症反応の増大、腸内フローラの変容を見いだした。自閉症モデルマウスとして知られているBTBRマウスでは、脳内ミクログリアの活性化、IL-6やTNF-alphaなどの炎症性サイトカインの上昇が見られた。また末梢の腸におけるサイトカイン発現では、TGF-betaの発現上昇が見られた。腸の免疫細胞のFACS解析の結果、BTBRマウスでは、マクロファージ、TH1, TH17の上昇、Treg, Th2の増加が見られた。腸内フローラの解析では、プロバイオティクスの減少と日和見細菌の増加が見られ、BTBRマウス腸での免疫系の異常が示唆された。またDSS処理による腸炎誘発の観察からもBTBRマウスにおける免疫反応の障害が示唆された。BTBRマウスの免疫細胞集団の変化は脾臓やリンパ節でも見られたため、共通の前駆細胞である造血幹細胞レベルでの異常が疑われた。よってB6マウスを放射線照射した後、BTBRマウスの骨髄を移植したところ、腸内フローラ、DSS反応性、免疫細胞集団等、ドナーマウスの表現型を呈した。以上の結果から、BTBRマウスは造血幹細胞レベルでの異常により、脳内、腸での免疫反応の異常を示すことが示唆される。


• Functional analysis of the novel adaptor protein, GAREM

  Konishi Hiroaki, TAKUMI Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Prefectural University of Hiroshima, 01 Apr. 2014 - 31 Mar. 2017

  GAREM1 (Grb2-associated regulator of Erk/MAPK1) is an adaptor protein that is involved in the EGF pathway. The nuclear localization of GAREM1 depends on the nuclear localization sequence (NLS), which is located at the N-terminal CABIT (cysteine-containing, all in Themis) domain. Here, we identified 14-3-3ε as a GAREM-binding protein, and its binding site is closely located to the NLS. Moreover, the binding of 14-3-3 had an effect on the nuclear localization of GAREM1. We suggest that the interplay between 14-3-3, the C-terminal SAM (sterile alpha motif) domain and CABIT domain might be responsible for the distribution of GAREM1 in mammalian cells. In contrast to GAREM1 that is expressed ubiquitously in human organs and cultured cells, GAREM2 is specifically expressed in the mouse, rat, and human brain. Moreover, GAREM2 does not possess the NLS sequence. To analyze the physiological functions of each GAREM subtype, we established and characterized a GAREM knockout (KO) mouse.


• Study for a development of a treatment of ASD with a clomosome duplicated ASD model mouse.

  Nagano Masatoshi, Takumi Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Nippon Medical School, 01 Apr. 2014 - 31 Mar. 2017

  We investigated a way of treatment for ASD by using a chromosome duplicated ASD model mouse (15q-dup). Because the serotonin content in the brain of 15q-dup mouse was decreased from neonatal period, we tried a treatment with a selective serotonin re-uptake inhibitor, fluoxetine, during 3 weeks after birth. The lower sociability and brain 5-HT level were ameliorated in adult 15q-dup mice, but their persistent behavior and repetitive behavior were not. Electrophysiological experiments revealed that 5-HT neurons had more hyperpolarized resting membrane potentials and smaller excitatory glutamatergic inputs in the dosal raphe nucleus in 15q-dup mice compared with the wildtype. In association with the serotonin restoration, neonatal FLX treatment also ameliorated these electrophysiological of 15q-dup mice. （Science Advances, in press）


• Common defects of RNA metabolism in motor neuron diseases SMA and ALS

  Tsuiji Hitomi, YAMANAKA Koji

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), 01 Apr. 2013 - 31 Mar. 2017

  Two motor neuron diseases, amyotrophic lateral sclerosis (ALS) and spinal muscular atrophy (SMA), are caused by distinct genes involved in RNA metabolism, TDP-43 and FUS, and SMN, respectively. However, whether there is a shared defective mechanism in RNA metabolism common to these two diseases remains unclear. Here, we show that TDP-43 and FUS localize in nuclear Gems through an association with SMN, and that all three proteins function in spliceosome maintenance. We also show that in ALS, Gems are lost, U snRNA levels are up-regulated and spliceosomal U snRNPs abnormally and extensively accumulate in motor neuron nuclei. This aberrant accumulation of U snRNAs in ALS motor neurons indicates both ALS and SMA motor neurons have defects in the spliceosome. These findings indicate that a profound loss of spliceosome integrity is a critical mechanism common to neurodegeneration in ALS and SMA, and may explain cell-type specific vulnerability of motor neurons.


• Establishment of next generation chromosome engineering

  Takumi Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research, Grant-in-Aid for Challenging Exploratory Research, Institute of Physical and Chemical Research, 01 Apr. 2014 - 31 Mar. 2016

  In the CRISPR-Cas9 system, a single guide RNA (sgRNA) directs the endonuclease Cas9 to a targeted DNA sequence for site-specific manipulation. The sgRNA has also been shown to play a role in activating the endonuclease activity of Cas9. This dual function of the sgRNA likely underlies observations that different sgRNAs have varying on-target activities. We evaluated the cleavage activities of 218 sgRNAs using in vitro Surveyor assays. We found that nucleotides at both PAM-distal and PAM-proximal region of the sgRNA are significantly correlated with on-target efficiency. Furthermore, we demonstrated that the genomic contexts of the targeted DNA, the GC percentage, and the secondary structure of sgRNA are critical factors contributing to cleavage efficiency.


• Integrative research on mood and clock

  Takumi Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A), Grant-in-Aid for Scientific Research (A), Institute of Physical and Chemical Research, 01 Apr. 2013 - 31 Mar. 2016

  Dysfunction of mood and the circadian clock together represent a large proportion of mental illness with adverse economic impact. Clinical and genetic evidence indicates that the two disorders are co-regulated but direct molecular evidence is lacking. We showed that GSK3β phosphorylation of PER2 is essential for co-regulation of mood and the circadian clock. In rodent depression models, we observed defective circadian rhythms associated with the loss of synchronized phosphorylation of GSK3β. Both molecular and behavioral phenotypes were restored by lithium, a mood stabilizer. GSK3β phosphorylation in PER2 was essential for both mood and circadian behaviors. Strikingly, Per2 mutant mice were resistant to both lithium and learned helplessness, a depression model. Together, these findings show that mood and the circadian clock share a coherent molecular mechanism and reveal the potential for synchronized drug and behavioral therapy of mood and circadian disorders.


• Brain Environment

  Takahashi Ryosuke, Kiyama Hiroshi, Yamanaka Koji, Higuchi Makoto, Uchiyama Yasuo, Hattori Nobutaka, Takumi Toru, Kawakami Hideshi, Urushitani Makoto

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Kyoto University, 01 Apr. 2011 - 31 Mar. 2016

  Project managing group members had served for management and planning of project meetings, international symposiums and research supports. The research meetings were held twice a year, and evaluated the progress of each project, collaborations, and the resource sharing. We organized the annual research workshop, and the international symposium with the distinguished oversea invitees from oversea, four times. We supported young investigators by research grant to generate transgenic mice and monoclonal antibodies. Two PhD students were awarded for the travel grant to attend the annual meeting of Society for Neuroscience in U.S. Many outreach activities for non-expert audience or high school students were performed to present our research activity. We supported the selected young investigators by a research grant required for their projects. Our achievements were open for discussion in the internet forum, and was summarized as pathway map, designated as “Brain Environment Map”.


• Altered RNA in neurons and its link to brain environment

  Takumi Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), 01 Apr. 2011 - 31 Mar. 2016

  Translocated in liposarcoma/Fused in sarcoma (TLS/FUS) is an RNA-binding protein that regulates the splicing pattern of mRNA transcripts and is known to cause a type of familial amyotrophic lateral sclerosis (ALS). In the absence of TLS, Mammalian enabled (Mena), an actin-regulatory protein and a target of TLS, undergoes preferential alternative splicing. We showed that the ablation of TLS dysregulates the subcellular location and functions of Mena. When TLS knockout mouse embryonic fibroblasts (MEFs) were transfected with wild type Mena, it no longer accumulated at focal adhesions and peripheral structures, whereas the localization of the alternatively spliced form was maintained. Additionally, Mena’s ability to suppress the motility of cells was lost in TLS knockout MEFs. Moreover, Mena failed to promote neurite outgrowth in TLS knockout primary neurons. Taken together, TLS is intimately involved in the local cytoskeletal dynamics surrounding Mena in both fibroblasts and neurons.


• Comprehensive Brain Science Network

  Kimura Minoru, Tanji Jun, Takada Masahiko, Nakamura Katsuki, Ohtsuka Toshihisa, Aoki Shigeki, Takao Hidemasa, Shimoji Keigo, Goto Masami, Yoshiura Takashi, Nakata Yasuhiro, Abe Osamu, Masumoto Tomohiko, Tokumaru Aya, Matsumura Akira, Kirino Eiji, Terada Hitoshi, Sato Noriko, Kasai Kiyoto, Hashimoto Ryota, Niwa Shin-ichi, Kato Tadafumi, Suzuki Michio, Shuji Iritani, Nemoto Kiyotaka, Tomita Hiroaki, Murayama Shigeo, Akatsu Hiroyasu, Takao Masaki, Saito Yuko, Bito Haruhiko, Yoshimura Yumiko, Matsuzaki Masanori, Furuta Toshiaki, Okado Haruo, Saito Izumu, Kaibuchi Kozo, Hasegawa Masato, Aiba Atsu, Shiina Nobuyuki, Igarashi Michihiro, Tomoki Nishioka, Watanabe Masahiko, Koike Masato, Sakagami Hiroyuki, Shigemoto Ryuichi, Fukazawa Yugo, Sakimura Kenji, Mori Hisashi, Mishina Masayoshi, Kobayashi Kazuto, Yanagawa Yuchio, Uemura Tadashi, Ishihara Takeshi, Nose Akinao, Iino Yuichi, Miyakawa Tsuyoshi, Takao Keizo, Mushiake Hajime, Katayama Norihiro, Tanaka Tetsu, Inoue Kazuhide, Okabe Shigeo, Kano Masanobu, Fujiyama Fumino, Isa Tadashi, Kageyama Ryoichiro, Fujita Ichiro, Yoshida Akira, Nishikawa Toru, Nukina Nobuyuki, Fukai Tomoki, Iwatsubo Takeshi, Yamamori Tetsuo, Okazawa Hitoshi, Tanaka Keiji, Kakigi Ryusuke, Tsuda Ichiro, Kitazawa Shigeru, Doya Kenji, Takahashi Ryosuke, Ikenaka Kazuhiro, Sobue Gen, Hasegawa Toshikazu, Ota Jun, Saitoe Minoru, Kadomatsu Kenji, Kida Satoshi, Manabe Toshiya, Tomita Taisuke, Iwata Atsushi, Murakami Ikuya, Tsutsui Ken-ichiro, Hanakawa Takashi, Hirai Hirokazu, Mima Tatsuya, Isomura Yoshikazu, Samejima Kazuyuki, Hoshi Eiji, Miyata Mariko, Yuzaki Michisuke, Tanaka Masaki, Fukata Masaki, Suzuki Kyoko, Kuba Hiroshi, Masu Masayuki, Kinoshita Makoto, Sugihara Izumi, Shirane Michiko, Yamamoto Nobuhiko, Nishijo Hisao, Nambu Atsushi, Takumi Toru, Yamashita Toshihide, Sakurai Takeshi, Tamamaki Nobuaki, Hata Yoshio, Harada Akihiro, Ozaki Norio, Sakai Katsuyuki, Kubo Yoshihiro, Nakazawa Takanobu, Tanaka Kenji, Takei Nobuyuki, Hitoshi Seiji, Takahiroa. Kato, Kato Fusao, Shirao Tomoaki, Taira Masato, Okano Hideyuki, Sekino Yuko, Okamoto Yasumasa, KOMATSU Hidehiko, Miyata Takaki, Takahashi Yoshiko, Nishida Shinya, Tominaga Makoto

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area), Center for Novel Science Initatives, National Institutes of Natural Sciences, 01 Apr. 2010 - 31 Mar. 2016

  The Comprehensive Brain Science Network (CSBN) provided individual researches supported by Grant-in-Aid for Scientific Research with cutting-edge resources and technologies, such as model animals, postmortem brain tissue, optical technologies for imaging and manipulation, virus vectors and more. Support covered researches on neuron-specific genes and molecules, synapse, network system, brain functions in disease states and neuro-computation. Special support was directed to collaboration between different fields. Workshop was held to have joint symposia among fields of peoples, to have a special session to let neuroscience community members share knowledge of how current researches are supported in Japan and discuss about future. Graduate students and postdoctoral fellows were supported to visit other laboratories of different field in Japan and abroad and learn disciplines, and award those who presented high quality work the CBSN Prize. These supports promoted break through from conventional approaches and publication of a number of papers with very high quality.


• Clinical application of physical activity time series: toward detection of early warning of depression

  YAMAMOTO Yoshiharu, TAKUMI Toru, YOSHIUCHI Kazuhiro, KITAJIMA Tsuyoshi, OSIMA Norihito, NAKAMURA Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A), Grant-in-Aid for Scientific Research (A), The University of Tokyo, 01 Apr. 2011 - 31 Mar. 2014

  In this study, we aimed at establishing a personalized early-warning-detection-system (sign-based Ecological Momentary Assessment) for mood disorders. We measured locomotor activity around clinical bifurcation points (e.g. relapse, exacerbation, and remission) and treatment, and then developed a behavioral biomarker for the diseases, characterizing motor abnormalities, specifically, alterations in behavioral organization and/or an increased intermittency of locomotor activity. In addition, we examined the underlying theoretical background of the behavioral organization by the mathematical modeling. Through these clinical and mathematical validations of our behavioral biomarker, we could obtain a firm basis for realization of early warning detection of the mood disorders, triggered by alteration in locomotor activity.


• Non-standard peptide-probe discovery

  SUGA Hiroaki

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Specially Promoted Research, Grant-in-Aid for Specially Promoted Research, The University of Tokyo, 2009 - 2013

  This research project aims at developing the genetic code reprogramming technology based on the flexizyme system and expressing natural product-like macrocyclic peptides from mRNA libraries. Moreover, an in vitro display system, referred to RaPID (Random nonstandard Peptide Integrated Discovery) system, has been established to apply for massive screenings of macrocyclic peptides generated by the above technology. This has enabled us to discover molecular probes against various drug targets, leading to a new era of chemical biology.


• The expression system specific for the circadian center

  TAKUMI Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research, Grant-in-Aid for Challenging Exploratory Research, Hiroshima University, 2011 - 2012

  The mammalian oscillator for circadian rhythm with a period of 24 hours is localized in the suprachiasmatic nucleus (SCN). Although the recent advance in genetic engineering methods provide different region-specific expression system, we have not obtained the specific expression system in SCN. The present study is to try to establish a system of SCN-specific expression using a combination of different promoters.


• 自閉症ヒト型モデルマウスを用いた社会性行動のシナプスパソロジー

  内匠 透

  日本学術振興会, 科学研究費助成事業 新学術領域研究(研究領域提案型), 新学術領域研究(研究領域提案型), 広島大学, 2011 - 2011

  自閉症でもシナプス異常が原因の一つとして提唱されている。また、CNV(コピー数変異)がこれまで考えられていた以上に多くの症例でみられることがわかってきた。応募者は、染色体工学的手法を用いて、ヒト染色体15q11-q13重複のモデルマウスの作製に成功した。15q11-q13重複は自閉症の細胞遺伝学的異常としてはもっとも頻度の高いもので、本モデルは、自閉症様行動を示すという表現型妥当性を示すだけでなく、自閉症の原因である染色体異常を患者と同じ型で有するという構成的妥当性をもみたす自閉症ヒト型モデルマウスである。本研究においては、本マウスのシナプス異常を中心に、シナプス異常がもたらす社会性行動異常のメカニズムを明らかにすることを目的とする。自閉症の原因の一つとして、近年シナプス関連遺伝子の異常及びその結果としてのシナプス異常が、ヒト遺伝学及びマウスモデルの結果から示唆されている。 自閉症ヒト型モデルマウスを用いて、細胞生物学的解析を行ったところ、patDp/+マウスの前頭皮質において、スパインの異常が見出された。本異常のとりわけin vivo状態での詳細な解析を行うために、2光子顕微鏡のセットアップを行っている。形成や消失傾向にあるスパインを記録することにより、定常状態のスパインのみならず、スパインの動的状態を記録している。また、PSD-95等の後シナプスマーカーを用いて染色することにより、シナプス可塑性との関連を考察した。


• 蜂蜜の神経薬理学的作用に関する統合的研究

  内匠 透, AKANMUMOSES ATANDA, AKANMU Moses Atanda, AKANMU MOSES ATANDA

  日本学術振興会, 科学研究費助成事業 特別研究員奨励費, 特別研究員奨励費, 広島大学, 2009 - 2011

  蜂蜜は植物の花蜜や植物甘味分泌物に由来する天然の甘味料である。蜂蜜は食品に容易に添加することができ、これまでに、蜂蜜に含まれるポリフェーノールやその他の天然化合物に起因する健康増進作用が報告されている。これら蜂蜜における天然成分の構成組成や抗酸化作用・神経保護作用は、それらの由来元である植物の種類や地理学的な位置に依存すると考えられる。そこで本研究では、ナイジェリアにおける産地の異なる3種の蜂蜜および日本産蜂蜜の抗酸化作用、神経保護作用、概日リズム、睡眠覚醒、抗ストレス作用に及ぼす影響を検討した。 蜂蜜のDPPHラジカル消去能に関しては、蜂蜜サンプルにおいて抗酸化作用が見られた。蜂蜜の神経保護作用および細胞増殖作用に関しては、ほとんどの蜂蜜は低濃度にて、血清非存在下(TIP:transferin,insulin,progesterone添加および非添加時)における細胞増殖作用を示した。蜂蜜の概日リズムに対する作用では、各蜂蜜サンプル(1,0.5%v/v)ではコントロールに比べRat-1細胞におけるper2プロモーター活性の位相変化が見られた。蜂蜜の抗ストレス作用では、各蜂蜜サンプル投与群では、拘束ストレスによるcorticosteroneレベルにおいて、精神安定剤であるdiazepamと同様の作用を示した。また、いくつかの蜂蜜サンプルにおいて、視床下部での拘束ストレスにより上昇する5-HTの抑制作用が見られた。蜂蜜の睡眠覚醒に対する作用においては、現在EEGデータを解析中である。以上のことから、蜂蜜は脳における中枢効果を有しており、それらはその産地によって異なることが示唆された。


• Integrative analyses of circadian rhythms based on environment

  TAKUMI Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B), Grant-in-Aid for Scientific Research (B), Hiroshima University, 2009 - 2011

  We use multiple high-throughput approaches including chromatin immunoprecipitation(ChIP)-based systematic analyses and DNA microarrays combined with bioinformatics, to generate genome-wide profiles of BMAL1 target genes. We reveal that, in addition to E-boxes, BMAL1 recognizes a consensus sequence CCAATG to elicit robust circadian expression. BMAL1 occupancy is found in more than 150 sites, including all known clock genes. Importantly, a significant proportion of BMAL1 targets include genes that encode central regulators of metabolic processes. The database generated in this study constitutes a useful resource to decipher the network of circadian gene control and its intimate links with several fundamental physiological functions

  Competitive research funding


• 概日センサーを介したリズム形成機構の解明

  内匠 透

  日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, 広島大学, 2009 - 2009

  概日リズムは、24時間で自転する地球環境に適応すべく生命が獲得した生理現象と考えられ、ほぼすべての生物に備わる基本的生命現象である。網膜で受容された光情報は、SCNに到達し、SCNニューロンは固有の概日時計を有するが、ニューロン間の同調によりSCNにおける中枢の概日時計が形成される。この中枢時計がロバストな概日リズムとして出力され、環境変化にダイナミックに対応し、生物個体の生存適応に必須の生命現象となる。本研究においては、光という環境情報を受容した後おこるSCNニューロン間での同調を概日セルセンサーとしてとらえ、SCN内の細胞間相互作用に必要な分子の同定を行う。またSCN部位特異的神経伝達阻害マウスを作製し、センサー間相関関係の機能的解析を行うことにより、概日センシング機構を解明する。生物が環境に適応する上で、時間のファクターは必須である。長期間にわたる生命現象の遺伝子発現は適切な時期に発現する必要があり、それらをリンクするものが時計遺伝子であるとも言える。マウスを生後恒常明(Light-light, LL)環境下で飼育した場合、成育後通常の明暗(Light-dark, LD)環境下に戻しても、暗期(マウスは夜行性)での明らかな行動異常(行動量の減少、一時的多動等)が見られることを発見した。これは、乳幼児期にLL条件下で飼育された結果、発達段階SCNでの細胞間情報伝達に24時間光刺激により何らかの異常をきたし、正常な中枢(SCN)時計が形成されなかったためと考えられ、この細胞間情報伝達に関与する分子あるいは機構が行動発達障害の引きがねとなるという仮説をたてることができる。研究代表者らは既にマウスをLL及びLD条件下で飼育したマウス由来のSCNを採取し、DNAマイクロアレイ法によって、LL条件下で発現変動している遺伝子群(26個)を同定した。

  Competitive research funding


• スパインにおけるタンパク質分解の生物学的意義の解明

  内匠 透

  日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, 広島大学, 2009 - 2009

  スパイン(棘突起)は神経細胞樹状突起上に存在する神経細胞に特徴的な構造物で、シナプス形成の場である。そのダイナミックな形態変化は、シナプス可塑性の構造的基盤と考えられる。また、神経細胞の細胞生物学的特徴の一つとして、樹状突起・スパイン内及び近傍にはポリリボゾームが存在し、局所タンパク質合成が知られており、シナプス可塑性の分子的基盤と考えられる。本研究では、神経細胞におけるユビキチンE3リガーゼUbe3aの標的基質を同定し、神経細胞でのUbe3aの機能を分子、細胞レベルで明らかにすることにより、樹状突起・スパインでの「ユビキチン・プロテアソーム系」の生理的意義を明らかにすることを目的とする。Ube3aの基質を明らかにするために、two-hybrid systemを用いてスクリーニングした結果えられた候補分子群の検討を行った。それぞれのタンパク質群の結合を哺乳類細胞及び初代神経細胞で確認した。また、初代神経細胞における発現を確認した。

  Competitive research funding


• ゲノム工学を用いて作製した自閉症マウスの解析による精神機能の分子的基盤研究

  内匠 透

  日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, 広島大学, 2008 - 2009

  ヒトゲノム計画が達成されたポストゲノム時代においても、脳機能においては、分子とシステムあるいは表現型との距離が今なお遠い。神経疾患においては、かなりの疾患関連遺伝子の単離、分子病態解明が進んだが、精神疾患においては、今なお臨床医学と基礎的脳科学の距離が離れており、アプローチすら困難な状況にある。研究代表者(内匠)は、最新の染色体工学的手法を用いて、ヒトの生物学異常としての染色体異常(インプリンティング領域でもあるヒト染色体15q11-13の重複)をマウスに構築し、精神行動異常(自閉症)の前向き遺伝学のためのファウンダーマウスを人工的に作製することに成功した。本マウスの解析により、ヒトモデルでもある父性重複ヘテロ(patDp/+)マウスには、社会的相互作用の障害、超音波啼鳴におけるコミュニケーション発達障害、常同様行動、ルーチン行動への執着性等の自閉症様行動がみられた。さらに、C57BL6にパッククロスしたマウスを用いて、行動解析を行った結果、既に見られていた行動結果と同様の、3-チャンバーテストにおける社会的相互作用の障害やBarnes迷路や水迷路の逆転学習における固執性等の自閉症様行動が種を超えて観察された。種を超えて自閉症様行動が見られることから、本モデルマウスの行動異常は種に依存したものではなく、重複領域にその原因があることが示唆された。また、本モデルマウスが、ヒト型自閉症モデルマウスとして有効であることを示している。

  Competitive research funding


• マウスフリームービング状態での神経核特異的遺伝子発現制御系の開発

  内匠 透

  日本学術振興会, 科学研究費助成事業 萌芽研究, 萌芽研究, (財)大阪バイオサイエンス研究所->広島大学, 2007 - 2008

  本申請研究では、既に確立したマウスフリームービング状態での視床下部視交叉上核(suprachiasmatic nucleus, SCN)の電気活動連続記録系(in vivo multiple unit neural activity, in vivo MUA)を発展させ、SCN特異的に遺伝子発現を制御する系を開発することを目的とする。In vivo MUAを用いて、SCNの電気活動の直接の出力が室傍核下部領域であることを示した。すなわち、SCNにおける電気活動の昼間に高い概日リズムは、SPZにおいては、夜に高い位相が逆転した概日リズムを示し、これらの逆位相関係の概日リズムは、分子時計が障害されているClock/Clock変異マウスにおいて、消失することを明らかにした。SPZは、睡眠覚醒、ホルモン分泌や体温調節等、恒常性維持に重要な働きをする視床下部内のいわば解剖学的ハブに位置しており、生物が環境に対して対応していく上(例えば昼行性か夜行性か)で必須な領域と考えられる。また、時計遺伝子プロモーターで制御されるルシフェラーゼモニターマウスを用いて、SCNスライスにおける単一細胞レベルでのリズム発現モニター系を確立した。数理解析の結果、SCN内に3つの位相クラスターが存在することを明らかにした。さらに、In vivo系モニターの検知に必要なCCDカメラの条件検討を行い、記録に必要な環境を得ることができた。

  Competitive research funding


• ミオシンVによるRNA顆粒輸送のメカニズムの研究

  内匠 透

  日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, (財)大阪バイオサイエンス研究所->広島大学, 2007 - 2008

  細胞内RNA顆粒は、RNAとRNA結合蛋白、翻訳関連蛋白他の蛋白からなり、遺伝子発現の転写後調節等の制御機能を有している。神経細胞に存在するRNA顆粒は、局所翻訳に関係し、シナプス可塑性の分子的基盤を考える上で重要である。研究代表者らは、RNA結合蛋白(TLS)を含むRNA顆粒が、アクチンモーターであるミオシンVにより、樹状突起からスパインに運ばれることを明らかにした。本研究では、ミオシンV及びTLSの相互作用蛋白を同定し、RNA顆粒の動態を経時的モニター系で解析することにより、神経樹状突起からスパインへのRNA輸送に関する「ナノシステム」の分子制御機構を明らかにすることを目的とする。酵母two-hybrid法により、ミオシンVに結合する蛋白をスクリーニングした結果、12個の候補遺伝子を同定し、海馬初代培養神経細胞系にて解析中である。これらの候補遺伝子のgain-of-function及びRNAiやミオシンVノックアウト(staggerer)マウスを用いたloss-of-functionの実験により、ミオシンVの輸送及びTLSを含むRNA顆粒の輸送との関連を明らかにする。また、網膜神経節細胞においては、TLSはカルシウム依存性にNMDA受容体であるNR1とmyosinVaと結合していることを明らかにした。TLSがカルシウム依存的なNR1スプライシングバリアントの輸送に関与することが示唆された。

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• Molecular mechanism of circadian oscillation by real-time rhythm monitoring

  TAKUMI Toru, TAKANO Atsuko

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B), Grant-in-Aid for Scientific Research (B), Osaka Bioscience Institute, 2006 - 2007

  The circadian expression of the mammalian clock genes is based on transcriptional feedback loops. Two bHLH-PAS domain-containing transcriptional activators, CLOCK and BMAL1, are known to regulate gene expression by interacting with a promoter element termed the E-box (CACGTG). The non-canonical E-boxed or E-box-like sequences have also been reported to be necessary for circadian oscillation. We found a new cis-element required for cell-autonomous circadian transcription of cock genes. This new element consists of a canonical E-box or a non-canonical E-box and an E-box-like sequence in tandem with the latter with a short interval, 6 base pairs, between them. We demonstrated that both E-box or E-box-like sequences are needed to generate cell-autonomous oscillation. We also verified that the spacing nucleotides with constant length between these 2 E-elements are crucial for robust oscillation. Thus, the direct repeat of the E-box-like elements is the minimal required element for the generation of cell-autonomous transcriptional oscillation of clock and clock-controlled genes. On the other hand, we developed in vivo monitoring of circadian timing in freely moving mice by recording multi-unit neural activity (MIJA). Suprachiasmatic nucleus (SCN), the mammalian circadian center, neural activity is tightly coupled to environmental photic input and anticorrelated with MUA rhythm in the subparaventricular zone (SPZ). In Clock mutant mice exhibiting attenuated circadian locomotor rhythmicity, MUA rhythmicity in the SCN and SPZ is similarly blunted. These results suggest that the SPZ plays a functional role in relaying circadian and photic signals to centers involved in generating behavioral activity.

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• 嗅神経回路形成におけるプロテアーゼを含む細胞外マトリックスに関する研究

  内匠 透

  日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, (財)大阪バイオサイエンス研究所, 2006 - 2007

  Fez2は、嗅上皮及び視床下部に特異的に発現するzinc-finger型転写因子である。本研究は、Fez2ノックアウトマウスのトランスクリプトーム解析で得られた細胞外マトリックス分子に焦点をあて、これら細胞外マトリックス分子のin vivoでの生理的機能を明らかにすることを目的とする。 匂いを受容する嗅神経は嗅上皮に存在し、脳の嗅球と神経連絡を形成し、発達期のみならず成体においても神経新生を行う神経として知られている。この神経伸張は性腺刺激ホルモン神経の伸張とも関与しており、ヒトのおけるこれらの神経伸張の異常はKallmann症候群を引き起こす。 Fez2欠損マウスの嗅神経では嗅球へ軸索が到達せず、これは軸索が胎生12.5日(E12.5)で嗅球表面の基底膜を貫通できないためであることが免疫組織染色で分かった。基底膜のモデルであるマトリゲル中で嗅上皮組織片を培養すると、Fez2欠損マウスの嗅神経では軸索が野生型より短くなり、軸索の基底膜分解能の低下が示唆された。さらに嗅上皮-嗅球共培養系において、基底膜を含む髄膜を除くと嗅神経の軸索伸張がレスキューされた。嗅上皮を用いたマイクロアレイの結果、セリンプロテアーゼを含む細胞外マトリックス分子の発現がFez2欠損マウスにおいてほぼ消失していることが分かった。 以上より本研究では、嗅神経軸索が嗅球へ到達する過程で、嗅球表面の基底膜を分解する際にFez2が重要な働きをすることが分かった。

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• 概日リズムのDECODEシステムの解明

  内匠 透, 高野 敦子

  日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, (財)大阪バイオサイエンス研究所, 2006 - 2007

  DNAマイクロアレイ法及びChIP (Chromatin immunoprecipitation)-chip(GeneChip)法によるヒト時間変動遺伝子群の解析を行うことにより、網羅的に遺伝子発現の時間的変動の分子機構、すなわち、時計転写因子の複合体ダイナミクス、また時間によりこれらがどう変動するかを明らかにしようとするものである。さらに、同定した時間変動遺伝子群に関しては、最近研究代表者らが確立したin vitro real-time oscillation monitoring system (IV-ROMS)を中心とした一連の時計遺伝子解析法を用いて、詳細な解析を行うことにより、ヒト時計遺伝子の転写制御「デコード」を目指す。 BMALl抗体を利用して、ラージスケールでヒト繊維芽細胞由来のゲノムDNAを用いてChIPを行うための種々条件検討を行い、さらにChIP-Chipのための準備を整えた。 一方、既に確立したIV-ROMSを用いた解析として、我々は、細胞自律的な時計遺伝子の概日転写に必要なシスエレメントを新しく同定した。この新規エレメントはE-box及びその6塩基下流にE-box様配列を有する新規EEエレメントである。我々は、EEエレメントの両方のE-boxが細胞自律的な振動を形成するのに必要であり、2つのE-box間の決まった長さ(6〜7塩基)が顕著な振動には決定的であることを明らかにした。さらに、ゲノムワイドの網羅的解析により、他の時間変動遺伝子の中にもこのEEエレメントが存在することを示した。以上の事から、概日転写制御には、従来のE-boxではなく、EEエレメントが重要であると示唆される。 また、NPAS2のプローモーター解析により、核内受容体ROR及びREV-ERVの結合部位RORエレメントがNPAS2の概日リズム発現に必要であることを明らかにした。

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• ゲノム工学を用いて作製した自閉症マウスの解析による精神機能の分子的基盤研究

  内匠 透, 高野 敦子, 中谷 仁

  日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, (財)大阪バイオサイエンス研究所, 2006 - 2007

  ヒト染色体15q11-13の重複は自閉症の細胞遺伝学的な異常としてもっともよく知られたものであるが、我々はCre-loxP系を基盤とした染色体工学的手法を用いて、マウス7番染色体相同領域(6.3Mb)の重複マウスの作製に成功した。本領域は、インプリンティング領域としても知られており、父性重複及び母性重複マウスと野生型マウスの比較検討を行った。 本マウス脳においては、重複された領域に存在する遺伝子群がインプリンティング依存的に量的増大を示した。 行動学的及び薬理学的解析の結果、父性重複マウスでは、ヒト自閉症様行動(社会性相互作用の低下、常同様行動、超音波啼鳴の発達遅延、不安傾向等)を示した。本モデルマウスは、ヒトと同様の表現型を示すだけでなく、ヒトと同様の構成的異常を備えたモデルである。 母性重複マウスではなく父性重複マウスが行動異常を示す原因の一つとして、snoRNA(MBI152)がセロトニン5-HT2c受容体のRNA editingを変化させることによりセロトニンシグナルに変化をきたす可能性を明らかにした。 さらに、マイクロダイアリスによる解析を行った。すなわち、セロトニン5-HT2c受容体アゴニストを側座核(Nucleus accumbens)に作用させるとドーパミンの放出が抑制されるが、父性重複マウスでは、このドーパミンの抑制がみられなかった。以上の事から、本父性重複マウスにおいては、セロトニン及びドーパミンシグナルの異常が見いだされた。

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• ゲノム工学を用いて作製した自閉症マウスの解析による精神機能の分子的基盤研究

  内匠 透, 阪本 昌美, 高野 敦子

  日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, (財)大阪バイオサイエンス研究所, 2005 - 2005

  精神疾患の分子的アプローチの困難さは的確なアッセイ系の欠如にある。本研究は、昨今のゲノム情報、最新の発生工学的手法を用いて、臨床の生物学的異常に基づくヒト染色体異常をマウスシンテニー染色体に導入することにより、ヒト精神疾患モデルマウスを確立・解析しようとするものである。具体的には、精神疾患の中でもより遺伝的寄与の高いと考えられる小児精神疾患である自閉症を標的としている。自閉症患者にみられるヒト染色体として15q11-13の重複がもっともよく知られているが、研究代表者らは、ヒト15q11-13に相当するマウス7Cの相当領域の両サイドにマウスES細胞でダブルターゲティングすることによりloxP配列を挿入し、Creレコンビナーゼにより重複をおこしたマウスを作製することに成功した。1)形態学的解析:脳のマクロ的、組織学的解析の結果、著明な形態学的変化はみられなかった。2)遺伝子発現解析:本ターゲット領域内の遺伝子群に関して、その発現を解析した。野生型、あるいは父性及び母性重複マウスの各組織での発現をQ-PCR法で体系的に解析した。一般的に、重複マウスでは、野生型に比べて発現量の上昇がみられ、また、父性及び母性遺伝子に関しては、そのインプリンティングに相応した発現量が観察された。一方、脳に発現する遺伝子群に関しては、それぞれのアダルトマウス脳組織での発現をin situ hybridization法により解析した。Ndn, Snrpn, Ube3a,Gabra5,Gabrb3はそれぞれ脳内での発現がみられ、やはり父性及び母性遺伝子に関しては、そのインプリンティングに相応した発現量が観察された。3)行動学的解析:網羅的な行動テストバッテリーによる探索・解析を行った。父性重複マウスでは、社会性の障害、常同運動、固執傾向、不安度の上昇等の行動異常が発見された。

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• 神経樹状突起における局所的蛋白合成によるシナプス可塑性の理解

  内匠 透

  日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, (財)大阪バイオサイエンス研究所, 2004 - 2004

  神経細胞樹状突起及び樹状突起棘では局所的蛋白合成が行われていることが示唆されており、この局所的蛋白合成が棘突起の形態的再構築さらにはシナプス可塑性の分子的基盤と考えられる。我々は、統合失調症の薬理モデルマウス(PCPマウス)のトランスクリプトーム解析で得られた候補遺伝子(RNA結合蛋白・TLS)を用いて、初代神経培養の細胞生物学的解析を行った結果、TLSが神経樹状突起に局在するRNA輸送蛋白であることやグルタミン酸シグナル依存的に興奮性シナプスを形成する棘突起へ移送することを明らかにした。またFUS/TLSのノックアウトマウスの培養海馬神経細胞では、棘突起の形態に異常が認められることから、FUS/TLSはmGluR5刺激依存的な棘突起の形態変化に重要な役割を果たしていると考えられる。さらに、我々はFUS/TLSがどのように棘突起のアクチン再構成やシナプスの形態変化に関わっているのかを明らかにするため、FUS/TLSとアクチンおよびアクチン細胞骨格依存性モーター蛋白の局所相互作用や、実際にFUS/TLS蛋白によって極性輸送されているmRNAの解析を行ったところ、アクチン安定化蛋白Nd1-L等の標的mRNAが同定された。シグナル依存性のTLSの神経細胞樹状突起棘への移行は、アクチン安定化蛋白Nd1-L mRNAの輸送を伴い、局所的蛋白合成を介してシナプス可塑性の基盤となることが予想される。

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• Functional analysis of an RNA-binding protein, TLS, in neuronal dendrites.

  TAKUMI Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Osaka Bioscience Institute, 2003 - 2004

  Neuronal dendrites, together with dendritic spines, exhibit enormously diverse structure. Selective targeting and local translation of mRNAs in dendritic spines have been implicated in synapse remodeling or synaptic plasticity. TLS (translocated in liposarcoma), previously identified as a component of hnRNP complexes, unexpectedly showed somatodendritic localization in mature hippocampal pyramidal neurons. TLS was translocated to dendrites as an RNA-protein complex and was recruited to dendrites not only via microtubules but also via actin filaments. In mature hippocampal pyramidal neurons, TLS accumulated in the spines at excitatory postsynapses upon mGluR5 activation, which was accompanied by an increased RNA content in dendrites. Consistent with the in vitro studies, TLS-null hippocampal pyramidal neurons exhibited abnormal spine morphology and lower spine density. We further demonstrated that TLS-null neuronal dendrites had decreased RNA cargo following mGluR-activation compared to wild-type dendrites and specifically identified an actin-stabilizing protein, Nd1-L, as a TLS-associated RNA cargo. TLS participates in mRNA sorting to the dendritic spines induced by mGluR5 activation and regulates spine morphology to stabilize the synaptic structure probably through actin-network, suggesting that TLS is necessary for spine maturation and the plasticity of excitatory postsynaptic sites.

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• 生物時計分子の核内移行と概日リズム

  内匠 透

  日本学術振興会, 科学研究費助成事業 特定領域研究, 特定領域研究, 神戸大学->(財)大阪バイオサイエンス研究所, 1999 - 2003

  培養細胞を用いた実験等から、CRYとPERとの核移行が、転写のフィードバック機構の本体を調節し、時間の決定機構となるという仮説をたて、時計分子の核移行の詳細な分子機構を検討した。CRY2には典型的な核移行シグナルNLSがカルボキシル端側に存在するが、CRY1には定型的NLSが存在しない。CRY1,CRY2をタグ付きGST融合蛋白として大腸菌にて発現、精製した蛋白をMH3T3培養細胞に注入することによりその核移行を検討した。CRY1,CRY2蛋白の細胞質内注入後、両者とも核内への移行がみられたが、WGA, Ran-GTP, G19V Ran-GTP等との共注入実験を行った所、同じく両者ともRan依存性に核移行することが明らかになった。さらに、CRY1,CRY2をそれぞれN末側C末側に二分割したCRY1N, CRY1C, CRY2N, CRY2Cを、同様に大腸菌を用いて蛋白を発現、精製し、細胞注入した結果、いずれもN末端側蛋白(CRY1N, CRY2N)は核移行を示さず、C端側(CRY1C, CRY2C)は核移行を観察した。セミインタクト細胞を用いたin vitro系では、CRY1C, CRY2Cともにimportinαβを介する核移行を行っていることが示された。NLS配列をもつCRY2Cの核移行はin vitro結合実験の結果からもimportinαβを介する核移行であることを明らかにした。さらに、CRY2のNLSの変異体では核移行が阻害されたことから、CRY2の核移行に必須であることを明らかにした。

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• Genome-wide analyses of genes predominantly expressed in the mouse neocortex

  TAKUMI Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Osaka Bioscience Institute, 2001 - 2002

  The mammalian cerebral neocortex occupies the largest area of the cerebral cortex and is cytoarchitectually composed of six layers (I-VI). Many of neurological diseases including the mental illnesses are due to the dysfunction of the neocortex. In spite of its functional importance, the neocortex is virtually an unexplored region by approaches of molecular biology including the genome-wide technology. The global screening using microarrays, combined with the systematic in situ hybridization, has provided us with several candidate genes that are expressed predominantly in the mouse neocortex. Among them, Fez, a zinc finger type transcription factor, was analyzed in further detail. No other genes in the neocortex have been known so far to have their expression the strongest in terms of specificity. Using two color in situ hybridization, we have shown that Fez is mainly expressed in the cortical layer V, not in the GABA neurons but in the pyramidal neurons, the projection neurons of the cerebral cortex. Molecules identified by our systematic analyses would be invaluable not only for molecular understanding of the development and function of the neocortex but also for the use of their promoters in the gene-manipulated animals as well as in conditional expression systems.

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• Measurement and Visualization of membrane potential by genetical manipul ati on.

  TAKUMI Toru, FUJII Ritsuko, KURIHARA Tatsuya, MANABE Toshiya

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B), Grant-in-Aid for Scientific Research (B), 神戸大学->(財)大阪バイオサイエンス研究所, 2000 - 2001

  In neuroscience it is important to analyze not only a single neuron but also network communication among multiple neurons. It is not easy for biochemists or molecular biologists to use conventional electrophysiology when they discuss electrophysiological phenomenon in the nervous system. We plan to develop an untouched voltage-sensing probe and establish a novel assay system, without using conventional electrophysiology, to analyze the neuronal functions, combined with voltagesensitive dyes and the imaging system. We developed electrophysiological analyses and assay systems with use of voltage-sensitive dyes. We recorded long-term potentiation (LTP) in CAl and CA3 of mouse hippocampus and made a study of mechanisms of pharmacological modification. Further, we succeeded the recording of LTP in mouse hippocampal neurons using multi-electrodes. The system we develop may be possible to be mechanized or robotized automatically. It is thus expected that the system should be contribute to not only research in basic medicine but also industrialization such as the systematic screening of the compounds. The system using voltagesensitive dyes may enable us to analyze interaction among multi-neurons, in addition to study a single neuron.

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• Molecular mechanism of mammalian biological rhythms

  TAKUMI Toru, OKAMURA Hitoshi

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Kobe University, 1999 - 2000

  Almost all organisms have their circadian rhythms with periods of approximately 24 hours. Circadian rhythms are generated in pacemaker cells and are entrained by environmental cues, such as light and temperature. The output of a circadian oscillation appears as locomotive activity, hormonal secretion, the steep-wake cycle, and many physiological functions. Recent molecular dissection has revealed that the feedback loop is the basic concept of the circadian oscillator, conserved across the species from Neurospora and Drosophila to mice. We have isolated a mammalian homologue of clock genes from the mouse brain. 1) To clarify the in vivo function of mammalian clock genes, we introduced the mammalian Per genes under Drosophila tim promoter into the null mutant per0 and have found in vivo rescue in some colonies. 2) We isolated human hPer3 cDNA and genome and found, compared with mPer3, VNTR (variable number of tandem repeats), SVs (splicing variants) and SNPs (single nucleotide polymorphisms). 3) We isolated a new zinc-finger type of transcription factor (Lot1) by differential display method. The Lot1 mRNA is highly expressed in SCN, the mammalian circadian center, during P1 to P10. 4) We showed expression of mPER2 and mPER3 protein in a circadian manner by serum shock to cultured N1H3T3 cells, but no rhythmic expression of TIM proteins.

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• 光による時計蛋白TIMELESSの分解機構の解明

  内匠 透, 岡村 均

  日本学術振興会, 科学研究費助成事業 特定領域研究(A), 特定領域研究(A), 神戸大学, 1999 - 1999

  生物リズム、特に約24時間周期の概日リズムはほとんどすべての生物にみられる基本的生命現象である。最近、我々は数種類の哺乳類時計遺伝子を単離、解析した。特に、ごく最近明らかにしたTIMELESS蛋白はショウジョウバエにおいて光による蛋白分解を受け、時計のリセット機構に関与することが知られている。本研究は、時計蛋白の分解機構を解析し、哺乳類概日系の光による同調機構を明らかにしようとするものである。 1、in vivoにおける時計分子の機能を明らかにするために、ショウジョウバエnull変異体を用いたin vivo rescueの実験を行った。ショウジョウバエnull変異体に哺乳類時計遺伝子(Per2,Per3)を導入することにより、行動リズムが回復することを確認し、生体におけるPERの核移行を検討した。 2、ヒトhPer3遺伝子を単離し、その中にVNTR(variable number of tendem repeat),SVs(Splicing variants),SNPs(single nucleotide polymorphisms)を明らかにした。 3、Differential display法を用いて、視交叉上核(SCN)由来のzinc-finger型転写因子(Lot1)を単離した。Lot1はP1からP10にかけてSCNにおいて強い発現が見られた。これはSCNニューロン間のシナプス形成の時期に一致しており、Lot1のシナプス形成における関与が示唆された。

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• 抗がん剤の薬物動態と投薬のタイミングの基盤となる生体リズムの制御機構

  内匠 透, 岡村 均

  日本学術振興会, 科学研究費助成事業 特定領域研究(A), 特定領域研究(A), 神戸大学, 1999 - 1999

  血圧、脈拍、体温や消化などの生理機能に概日リズムがあることから、薬物動態、すなわち投与薬物の吸収、分布、代謝、排泄が生体リズムの影響を受けることが知られている。癌の治療を考える上で、抗がん剤の時間薬理学的考察はその重要性が指摘されてはいるものの、抗がん剤の開発や他の治療法に比べて研究が進んでいないのが現状である。本研究は、薬物動態の日周リズムを考慮することにより、抗がん剤の効果を増強し、副作用を軽減することを目的とした時間薬物治療をめざして、ヒトを含めた高等動物において、概日リズムを分子のレベルで明らかにしようとするものである。 1、in vivoにおける時計分子の機能を明らかにするために、ショウジョウバエnull変異体を用いたin vivo rescueの実験を行った。ショウジョウバエnull変異体に哺乳類時計遺伝子(Per2,Per3)を導入することにより、行動リズムが回復することを確認し、生体におけるPERの核移行を検討した。 2、ヒトhPer3遺伝子を単離し、その中にVNTR(variable number of tendemrepeat),SVs(Splicing variants),SNPs(single nucleotide polymorphisms)を明らかにした。 3、Differential display法を用いて、視交叉上核(SCN)由来のzinc-finger型転写因子(Lot1)を単離した。Lot1はP1からP10にかけてSCNにおいて強い発現が見られた。これはSCNニューロン間のシナプス形成の時期に一致しており、Lot1のシナプス形成における関与が示唆された。

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• Generation of transgenic mice with arrythmia expressing a dominant-negative Isk

  TAKUMI Toru, SHIBANO Toshiro, MATSUDA Hiroko

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B), Grant-in-Aid for Scientific Research (B), Kobe University, 1998 - 1999

  To understand physiological function of the Isk channel in heart, we investigated behavior of the Isk channels in three different ways ; in mammalian cultured cells, in isolated ventricular myocytes and in the heart of transgenic mice. (1)In COS7 cells transfected, Isk was associated with KvLQT1 and HERG. The rapidly activating potassium current was abolished by cotransfection of D77N dominant negative Isk and HERG cDNA. (2)In isolated ventricular myocytes, two types of outward KィイD2+ィエD2 currents, the rapidly activating potassium channel (which corresponds to IKr) and the slowly activating potassium channels (which corresponds to IKs), are identified. The outward KィイD2+ィエD2 currents were inhibited by the antisense Isk oligonucleotide or D77N dominant negative Isk mutant. The action potential duration in ventricular myocyte was significantly prolonged by introduction of D77N dominant negative mutant. (3)Transgenic mice expressing a dominant-negative D77N Isk in heart were generated under the control of α-myosin heavy chain promoter. The introduced Isk was strongly expressed in the heart. The electrocardiogram of the mice and electrophysiology using the isolated ventricular myocytes from the mice have been analyzed.

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• サーカディアンリズム形成に伴う時計遺伝子の発現とその制御機構

  岡村 均, 山口 瞬, 重吉 康史, 内匠 透

  日本学術振興会, 科学研究費助成事業 特定領域研究(A), 特定領域研究(A), 神戸大学, 1998 - 1998

  先年度発見したmPer1に続き、ショウジョウバエperiodホモローグであるマウス時計遺伝子mPer2,mPer3を世界で初めてのクローニングした。mPer2は、mPer1と同じく、リズム中枢のある視交叉上核で昼は強く発現し、夜はほとんど発現しないが、そのピーク時間は約4時間ずれていた(Genes Cells,1998)。もちろん、恒常暗条件下でもこのリズミック名発現は検出された。続いて発見したmPer3の発現パターンは、明暗条件下でも、恒常暗条件下でも、昼に広くなだらかなピークのある発現を示し、夜でも発現量が減るが確実に発現し、mPer1,mPer2と異なるパターンであった(EMBOJ.,1998)。 視交叉上核の発生時期に発現する転写因子をDIFFERENTIAL mRNA DISPLAY法にて検索した。その結果、生後10日間のみ発現するDNA結合ドメインを持つ転写因子lot1を単離した。このmRNAの発現は恒常暗条件下でも、著明なサーカディアンリズムを示した。この転写因子は、視交叉上核以外の脳部位にも強く発現し、それら部位の発達に関与していることが示唆された。詳細な高感度の検出法では、発生終了後もこの遺伝子発現は続くが、その意義は不明である。ごく最近、mPer1とへテロダイマーを形成する哺乳類Timelessのクローニングにも成功した(Genes Ce11s,1999).このヘテロダイマー形成は核の中でのみ見つかっており、今後の検討が待たれる。

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• 時計遺伝子のリズム位相制御機構

  岡村 均, 山口 瞬, 重吉 康史, 内匠 透

  日本学術振興会, 科学研究費助成事業 特定領域研究(A), 特定領域研究(A), 神戸大学, 1998 - 1998

  クローニングしたmPER3はmPERl,mPER2よりやや小さく、1115アミノ酸からなる蛋白質であった。ショウジョウバエdPerの特徴とされる、蛋白質相互の結合に関与するPASドメイン(PAS-A,PAS-Bからなる)は、mPER1,mPER2と同様よく保存されていた。また、ショウジョウバエ時計遺伝子dPerを細胞質にとどめ、核内移行を抑制するcytoplasmic localization domain(CLD)構造が保存されていることも、大きな特徴である。mPER3には他のmIPER1,mPER2に認められないnuclear localizationsignal(NLS)がある。視交叉上核においては、明暗条件下、恒常暗条件下でも24時間リズムを示す。そのリズムは、CT4-8にかけて高く、mPERl,mPER2と同様、 「昼時計」型であるが、nPERl,mPER2と異なり、夜にもかなり発現しており、昼夜での振幅は小さい。 我々は、哺乳類のmPERの発現にもtimeless遺伝子が重要かどうかを検索するため、mTIMのクローニングを行った。mTIMは1197アミノ酸からなる蛋白質で、dTIMと4つのドメインで相同性が高かったが、全く異なる配列が、挿入されていた。mTIMとdTIMの分子構造を比較すると、確かにmTIM分子はPERとの結合領域のうちPAS-Aと結合する部分は保存されているが、PAS-B/CLDと結合する部分には新たなアミノ酸の挿入があり、dTIM分子が細胞質内にいてPER/TIMの核移行を妨げる重要な性質であるdTIMのCLDはほとんどmTIMでは保存されていない。このことは、mTIMの3カ所のNLSに引きずられ、mTIMは通常の核蛋白として、核の中に直ぐ入ってしまい、TIMが哺乳類では核移行のregulation stepを司る役割を失っている事を示唆している。

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• 松果体メラトニンリズムの神経栄養因子による制御機構

  藤本 悦子, 重吉 康史, 内匠 透, 岡村 均

  日本学術振興会, 科学研究費助成事業 基盤研究(C), 基盤研究(C), 神戸大学, 1997 - 1998

  脊椎動物における概日リズム(circadian rhythm)をつかさどる時計(circadian clock)の所在が明らかになったのは、鳥類の松果体である。これに対し、哺乳類松果体は、光受容能が全く失われており、メラトニン合成の日内リズムは完全に上頚神経節由来の交感神経系の支配を受けており、上頚神経節除去にてこのリズムは消失する。今回、ラット松果体の交感神経を切断し、2週間後の松果体を検索した。切断は、チロシンヒドロキシラーゼの免疫組織化学をおこない、上頚神経節からのノルアドレナリン線維の投射が消失していることで、確認している。電子顕微鏡にても神経終末構造の消失を確認している。松果体でのNGF,b-FGF,GDNF発現を検索したが、発現の大幅な変動は認められなかった。次に、切断時期に変動する成長因子関連遺伝子や新規因子を見つけるため、PCR法を応用したdifferential mRNA dispalyにて検索した。このうち、成長因子や成長因子受容体及び、そのホモロジー遺伝子を検索する。現在、6種類の候補遺伝子を得て、クローニング中である。また、シュワン細胞の基底板管を用いた末梢神経の再生のモデルを用いてbFGFの効果を検索した。bFGFは神経軸索に直接に働き再生を促進することを確認した。

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• Molecular analysis of the mammalian circulation rhythm

  OKAMURA Hitoshi, YAMAGUCHI Shun, SHIGEYOSHI Yadufumi, TAKUMI Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B), Grant-in-Aid for Scientific Research (B), Kobe University, 1997 - 1998

  A new mammalian period complementary DNA, mPer2, has been isolated from mouse brain. The amino acid sequence of mPer2, encoding PAS-domain (PAS, a dimerization domain present in PER, ARNT, SIM), is similar to mPer1 and Drosophila Period (dPer), indicating that mPer2 is a family with mPer1, a homologue of dPer. The mPer2 mRNA is predominantly expressed in the suprachiasmatic nucleus (SCN), the center of biological clocks in mammals. A robust circadian rhythmic expression in the SCN in constant darkness supports mPer2 to be a rhythmic pacemaker. The peak of its expression is at evening in constant dark condition and light-off time in light-dark condition, indicating that expression pattern of mPer2 mRNA is different from day-type clock mPer1. A new member of the mammalian period gene family, mPer3, was isolated and its expression pattern characterized in the mouse brain. Like mPer1, mPer2 and Drosophila Period, mPer3 has a dimerization domain (PAS) and a cytoplasmic localization domain (OLD). mPer3 transcripts showed a clear circadian rhythm in the suprachiasmatic nucleus (SCN). Expression of mPer3 was not induced by exposure to light at any phase of the clock, distinguishing this gene from mPer1 and mPer2. Cycling expression of mPer3 was also found outside the SCN in the organum vasculosum lamina terminalis (OVLT), a potentially key region regulating rhythmic gonadotropin production and a pyrogen-induced febrile phenomena. Thus, mPer3 may contribute to pacemaker functions both inside and outside the SCN. To underst how light might entrain a mammalian circadian clock, we examined the effects of light on mPer1, a sequence homolog of Drosophila per, that exhibits robust rhythmic expression in the SCN. mPer1 is rapidly induced by short duration exposure to light at levels sufficient to reset the clock, and dose response curves reveal that mPer1 induction shows both reciprocity and a strong correlation with phase shifting of the overt rhythm. Thus in both the phasing of dark expression and the response to light mPer1 is similar to Neurospora clock gene frq. Within the SCN there appears to be localization of the induction phenomenon of mPer1, suggesting two types of clocks (light-responsive and light-unresponsive) being localized in this circadian center.

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• 細胞膜チャネル分子からのアプローチによるリズム発振細胞における情報伝達機構

  内匠 透

  日本学術振興会, 科学研究費助成事業 奨励研究(A), 奨励研究(A), 神戸大学, 1997 - 1998

  哺乳類概日時計の分子機構は一昨年の哺乳類時計遺伝子の単離(Per1,Pcr2)により、哺乳類においても、ショウジョウバエで知られている基本的機構が保存されている可能性が示唆されている。 我々は、一次構造上PAS、CLDドメインがありショウジョウバエPerとも相同性を有するPeriodファミリーメンバーのーつであるmPer3をマウス脳より単離した。mPer3mRNAは、全身組織に分布するが、脳内では哺乳類リズムセンターである脳視床下部視交叉上核(SCN)に発現し、その発現は概日リズム(昼間高く夜間低い)を示した。mPer3mRNAはSCN以外の視床下部領域にも発現し、特に終板器官においてはSCN型のリズミックな発現が見られ、体温調節リズムやゴナドトロピンリズムとの関連が注目される。mPer3はいかなる時期における光刺激に対しても不応性であり、哺乳類における最初の光非依存性のperiod遺伝子であると結論した。 さらに、in silicoスクリーニングにより、ショウジョウバエにおける第二の時計遺伝子timelessの哺乳類ホモログ断片を発見し、マウス脳よりmTim cDNAを単離した。mTim mRNAのSCNにおける発現には著明な概日リズムが見られず、光刺激による転写物の誘導もみられなかったが、網膜においては特に明暗条件下でその発現に概日リズムが見られた。また、mTIMはmPER1と結合することを生化学的に証明した。

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• Molecular Mechanism in Regulation of Inwardly Rectifying Potassium Channel and its Functional Disorder

  KURACHI Yoshisa, TANEMOTO Masayuki, INANOBE Atsuhi, HORIO Yoshiyuki

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A), Grant-in-Aid for Scientific Research (A), Osaka University, 1995 - 1998

  (1) ATP sensitive potassium channels A) We cloned a new sulfonylurea receptor(SUR2b) and revealed it forms a smooth muscle type ATP sensitive K+channel with Kir6.1. ATP sensitive K+channel is reported to be regulated by intracellular nucleotides and K+channel openers. We found out that the regulation of the ATP sensitive K+channel is different according to the difference of the channel forming units. B) We made several chimeras between Kir6.1 and Kir6.2, and by using these chimeras we revealed that the pore-forming region of the channel is responsible for determining channel conductance. Furthermore we found out the amino acids Ser (113), Ile (114), His (115), and Val (138) are major determinant of channel conductance of Kir6.2. (2) G-protein regulated potassium channels A) We revealed G-protein regulated K+channel of dopaminergic neurons in substancia nigra (SN) is composed from channel subunits, Kir3.2a and Kir3.2c. We also revealed SAP97, one of the members of anchering proteins which have PSD domains, is also expressed with these K+ channels. B) We showed Kir3.1 and Kir3.4 are expressed on the membrane of secretary vesicles in TSH secreting cells of pituitary body. By stimulating these cells with agonists, functional K+channels is expressed on the plasma membrane after fusion of secretary vesicles to the plasma membrane. C) We cloned a new subunit of G-protein regulated K+channel, Kir3.lb. This clone is a spliced variant of Kir3.la (former reported as Kir3.1). Checking the function of this clone, we showed strong evidence that C-terminal portion of Kir3.1 is indespensable for the channel activation by G-protein. (3) other potassium channels A) Kir2.Os are expressed on different cells in olfactory bulb (GB). This cell-specific expression of each channel units might indicate the functional specificity of them in the olfactory sensory system. B) In CNS, Kir4.1 (Kirl.2) was distributed especially on the basolateral membrane of epithelium of inner lymph follicle, glial cells, and ependymal cells. In retina, it is expressed on Muller cells. These specific distribution of this channel reflects a functional importance of Kir4.1 in handling potassium ions between different ional milieus. C) We also cloned new four membrane spanning K+channel (CTBAK) from heart cDNA library.

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• 概日リズムの形成及び同調に伴う遺伝子発現とその制御機構

  岡村 均, 重吉 康史, 内匠 透

  日本学術振興会, 科学研究費助成事業 重点領域研究, 重点領域研究, 神戸大学, 1997 - 1997

  哺乳類の体内時計に関しては、1970年代に体内時計のマスタークロックが視床下部視交叉上核に局在するという発見以来、この核の機能解析を中心に研究が進んできたが、時計自身の物質的な根拠に乏しいため、その根本的なメカニズムに迫るには限界があった。この状況で我々は程とともにショウジョウバエ時計遺伝子Periodの哺乳類ホモローグをクローニングし、視交叉上核での時間特異的発現を報告した(Nature,1997)。我々はこの哺乳類時計遺伝子mPer1が、哺乳類における重要な時計遺伝子であることを数々の方法で検索中であるが、本重点研究においては、この時計遺伝子哺乳類mPer1の発現が、光による行動リズムの位相変位にどのように関与するかを検索した。フリーラン条件下で、mPer1 mRNAの視交叉上核での発現を検索すると、約23時間間隔の発現変動が数周期にわたり観察された。mPer1 mRNAは視交叉上核には主観的暗期には全く認められないが、主観的明期に入ると上昇し始め明期開始後4時間でピークに達する。その変動幅は5-10倍あり、4周期後も全く低下傾向は示さなかった。この事は、行動リズムと同様、視交叉上核でのmPer1の発振メカニズムは恒常暗条件にても安定であることを示している。また、主観的暗期に光パルスを与えると、視交叉上核でmPer1の遺伝子が誘導された。また、この発現は、光量のlog値に比例して上昇した。この変動量は光による行動の位相変位と全く相関していた。この事は、mPer1の発現程度と行動位相変位の程度が全く相関する事を示しており、mPer1発現が位相変位に関与していることをさらに想定させる。

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• Analysis of the polyamine-binding sites in inwardly rectifying K^+ channels.

  YAMADA Mitsuhiko, INANOBE Astushi, ISOMOTO Shojiro, TAKUMI Toru

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B), Grant-in-Aid for Scientific Research (B), OSAKA UNIVERSITY, 1996 - 1997

  We analyzed the Mg^<2+>/polyamine sensitivity of the IRK2 (Kir2.2) channel heterologously expressed in HEK293T cells. IRK2 (Kir2.2) is believed to be the subunit of the cardiac inwardly-rectifying K^+ channel, I_. The IRK2 channels showed strong inward rectification in the cell-attached configuration of the patch-clamp method. However, they gradually lost the inward rectification in the inside-out patch membranes whose intracellular side was continuously perfused with Mg^<2+>-free solution. Mg^<2+> and spermine spplied to the internal side of the patch membrane suppressed the outward channel currents at the potential 40 mV positive to the potassium equilibration potential (E_K) with the IC_<50> of 10muM and 3 nM,respectively. Because millimolar and submillimolar Mg^<2+> and polyamine are known to exist in cytosol of most cell types, the Mg^<2+>/Polyamine block is likely to underlie the inward rectification of IRK2 channels in intact cells. Mg^<2+> caused the instantaneous rectification by inducing the subconducting level of the channel, while spermine time-dependently suppressed the open probability of the channel at potentials positive to E_K. When Mg^<2+> was further applied to the channel in the presence of spermine, the inward rectification of the channel was paradoxically attenuated compared with that in the presence of spermine alone. This phenomenon was well explained by the model in which Mg^<2+> and spermine compete with each other through binding the same binding site (s) on the channel. These data (1) indicate that the physiological inward rectification of the cardiac I_ channel is determined through such competitive binding of intracellular Mg^<2+>/polyamine to the channel and (2) raise a possibility that artificial polyvalent cations introduced into cardiocytes might be utilized to pharmacologically modulate the strong inward rectification of the channels induced by intracellular polyamines.

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• 内向き整流特性カリウムチャネルの神経伝達調節と可塑性における役割

  倉智 嘉久, 内匠 透

  日本学術振興会, 科学研究費助成事業 重点領域研究, 重点領域研究, 大阪大学, 1996 - 1996

  内向き整流カリウムチャネルスーパーファミリーの中枢神経系における分子機能解析を目的としてIRK1,IRK2,IRK3についてin situ hybridizationによって脳内の詳細な分布を調べそれぞれが特徴ある発現様式を持っていることを明らかとした。また、脳の内向き整流カリウムチャネルの調節機構を調べるため、脳mRNAをツメガエル卵母細胞に発現させてパッチクランプ法により解析した。脳の内向き整流カリウムチャネルはcAMP,cGMPによって抑制されることが明かとなった。この機構によってNOを含む各種のニューロトランスミッターが脳の内向き整流カリウムチャネル活性を抑制して、膜電位を上昇させ細胞の興奮性を上げることに働くと考えられ、新たな神経細胞興奮性の調節機構と考えられる。また、免疫組織化学的手法によりGIRK1はプレシナプスに存在することを明らかとした。このプレシナプスにおけるGIRK1の存在はこれまで全く考えられておらず、脳におけるG蛋白質制御カリウムチャネルの機能が単純なシナプス後抑制では無いことを示唆するものである。ATP依存性カリウムチャネルのK_-2の分布については既にグリア細胞にその発現がみられることを明らかとしていたが、網膜のグリア細胞であるMuller細胞にクラスター状に発現していること、Muller細胞では恐らくSAP97がこのチャネルの集積に関与していること、また、HEK細胞にSAP97を共発現させるとSAP97がK_-2に結合すること、SAP97がK_-2の膜上での発現を増加させることを明らかとした。ゲノム遺伝子のクローニングについても既にIRK3 geneを単離していたがさらにK_-2、GIRK1 geneを単離した。IRK3についてはIRK3を不活性化した遺伝子と相同組み替えをおこしたES細胞の作成に成功した。また、脳cDNAのスクリーニングによって新たな膜2回貫通型カリウムチャネルサブユニットGIRK2B,GIRK1B,uKATP,BIRをクローン化した。

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• 標的遺伝子組換えによるグリア細胞特異的内向き整流カリウムチャネルの解析

  内匠 透

  日本学術振興会, 科学研究費助成事業 奨励研究(A), 奨励研究(A), 大阪大学, 1996 - 1996

  1)K_-2チャネルゲノム遺伝子の構造解析:マウスゲノムライブラリーよりK_-2ゲノム遺伝子を単離し、その構造解析を行った。K_-2遺伝子はエクソン1及び2からなり、蛋白コーディング領域にはイントロンが存在しないことを明らかにした。現在、エクソン1上流のプロモーター領域の解析、さらにK_-2遺伝子の染色体マッピングを行っている。 2)K_-2チャネルを発現しないノックアウトマウスの作成:上記遺伝子を利用してターゲティングベクターを作製した。今後は、このベクターをES細胞に導入し相同組み換えの生じている細胞をサザングロット法により選別する。得られた細胞を宿主胚盤胞に注入し、偽妊娠雌マウスの子宮内に移植することによりキメラマウスを得る。これらのキメラマウスを使い、相同組み換えが生じているノックアウトマウスを得る。

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• Molecular Analysis of Inward Rectifier K^+channels in CNS

  KURACHI Yoshihisa, ISOMOTO Shoujiro, YAMADA Mitsuhiko, TAKUMI Toru, HORIO Yoshiyuki, DAVID Chella S

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for international Scientific Research, Grant-in-Aid for international Scientific Research, Osaka University, 1995 - 1996

  To analyze inwardly rectifying potassium channels (IRKs) in the brain, we have cloned cDNAs of GIRK1B and GIRK2B.These channels may participate in the reguration of GTP-binding protein-regulated potassium channels (K_G). We also cloned SUR2B (sulfonyl urea receptor 2B) cDNA which is a subunit of the ATP-regulated potassium channel expressed in vascular smooth muscle. Expression of SUR2B was ubiquitous and was also found in the brain. The genes for IRK3, K_-2, and GIRK1 were isolated. These genes will be used for knockout studies. To investigate the distribution of IRLs in the central nervous system, we performed in situ hybridization. These IRK channels were specifically expressed in special sets of neurons. Large amount of IRK2 mRNA was detected in the granular cell later of cerebellum, on the other hand, IRK3 mRNA was localized in the forebrain. Immunohistochemical studies showed that GIRK1 was expressed in presynapses in the paraventricular nubleus. We have previously demonstrated that K_-2 was predominantly expressed in glial cells. Using anti-K_-2 antibody, we further analized the distribution of K_-2, and found that K_-2 was expressed in Muller cells, retinal glial cells, and marginal cells, which are thought to contribute elevation of endolymph potential of inner ear. In these cells, K_-2 may have a key role inthe transport of K^+ from cells to cells. We also found that K_-2 was clustered in the membrane of Muller cells. K_-2 was associated with PSD-95/SAP90 and clustered when K_-2 was co-expressed with PSD-95/SAP90 in HEK293T cells. We also found that teh expression of PSD-95/SAP90 family proteins enhanced tha K_-2 current in HEK293T cells. Electrophysiological properties of inwardly rectifying potassium channels were studied to investigate the function and regulation of these channels in the brain. We found that inward rectification of K_G channels and cloned IRK2 channels was caused by intracellular polyamines. We also found that the binding of more than two molecules (could be four) of GTP-binding protein betagamma heterodimer to a single K_G channel was needed to open the channel. As a K_G channel was composed of four subunits of GIRK channels, our data suggested that each subunit binds one molecule of betagamma heterodimer.

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• Molecular cloning of ATP-sensitive K^+ channels and development of the method evaluating K^+ channel openers.

  KURACHI Yoshihisa, YAMADA Mitsuhiko, TAKUMI Toru, HORI Masatsugu

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (A), Grant-in-Aid for Scientific Research (A), Osaka University, 1994 - 1996

  ATP-sensitive potassium channels (K_), which represent a family of K^+ channels inhibited by intracellular ATP,have been found in various tissues including heart, pancreatic beta-cells, and smooth muscle. K_ in different tissues exibit considerable variation in responce to K^+ channel openers (KCO). In the present study we investigated the molecular difference of K_ channels and establilsh a in vitro screening system to categorize KCOs and also for developping new KCOs. K_ consists of two different subunits, K^+ channel subunit and sulfonylurea receptor (SUR) subunit. We have cloned BIR/Kir6.2 and uK_/Kir6.1, which are K^+ channel subunits. Then we isolated SUR2A and SUR2B cDNAa for SUR subunits. SUR2B was considered as a splice variant of SUR2A and had a very similar sequence to that of SUR1 in its carboxy terminal 42 amino acid residues. We expressed these cDNAs in HEK293T cells and assayd the cells with patch clamp method. When SUR2A and BIR were transfected in HEK cells, very similar channels to those in cardiac muscle were constructed. On the other hand, a combination of SUR2B and uK_ represents a K_, whose properties were almost same as those of vascular smooth muscle K_. Pinacidil could activate both SUR2A/Bir and SUR2B/uK_ channels. Small amount of diazoxide was able to open the SUR2B/uK_ channels but not those of SUR2A/Bir. Thus, using cloned K_ channel subunits and HEK293T cells, we reconstituted K_ channels of cardiac and vascular smooth muscle and evaluated the effects of KCOs on these channels. This in vitro system will be used to categorize KCOs and also develop new drugs.

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• 内向き整流特性カリウムチャネルの神経伝達調節と可塑性における役割

  倉智 嘉久, 山田 充彦, 内匠 透, 堀尾 嘉幸

  日本学術振興会, 科学研究費助成事業 重点領域研究, 重点領域研究, 大阪大学, 1995 - 1995

  中枢神経系における内向き整流性カリウムチャネルの分子機構を明らかにするために、マウス脳よりまったくあたらしい2回膜貫通型内向き整流性カリウムチャネルcDNA(K_-2,the second type of inward rectifying K^+ channel with an ATP-binding domain)をクローニングした。アフリカツメガエル卵母細胞発現系でK_-2チャネルは、古典的な内向き整流特性を示すカリウム電流を示した。in situ hybridization法による解析の結果、K_-2mRNAは脳幹部神経核とともに、脳梁や小脳白質などのグリア細胞に強く発現していた。さらに、網膜のミューラー細胞やアストロサイト初代培養にもK_-2チャネルが発現しており、グリア細胞におけるK^+-buffering systemに関与していると考えられる。また、免疫組織化学の結果、K_-2チャネルは腎臓遠位尿細管細胞や内耳血管条基底細胞の基底外側に発現していた。以上のことからK_-2チャネルは神経系グリア細胞や腎臓、内耳において細胞内外のK^+イオンの輸送に関与していると示唆される。現在、特にグリア細胞K^+-buffering systemにおけるK_-2チャネルの生理的役割を明らかにするために、ES細胞を用いた発生工学的手法によりK_-2チャネルのノックアウトマウスの作成を試みている。

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• 中枢神経系におけるATP感受性K^+チャネルの分子機能解析

  内匠 透

  日本学術振興会, 科学研究費助成事業 奨励研究(A), 奨励研究(A), 大阪大学, 1995 - 1995

  中枢神経系における内向き整流性カリウムチャネルの分子機構を明らかにするために、マウス脳よりまったくあたらしい2回膜貫通型内向き整流性カリウムチャネルcDNA(KAB-2,the second type of inward rectifying K^+ channel with an ATP-binding domain)をクローニングした。アフリカツメガエル卵母細胞発現系でKAB-2チャネルは、古典的な内向き整流特性を示すカリウム電流を示した。in situ hybridization法による解析の結果、KAB-2mRNAは脳幹部神経核とともに、脳梁や小脳白質などのグリア細胞に強く発現していた。免疫組織化学の結果、アストロサイト初代培養にKAB-2チャネルが発現した。また、KAB-2チャネルは胎児期脳には発現が少なく生後1週でその発現が著しく増加し、生後2週で成体レベルに達する。この発現パターンは生後脳内の神経ネットワークの形成とともにグリア細胞の増殖していくことと一致する。以上のことから、KAB-2チャネルはグリア細胞におけるK^+-buffering systemに関与していると強く示唆される。現在、生体内、特にグリア細胞K^+-buffering systemにおけるKAB-2チャネルの生理的役割を明らかにするために、ES細胞を用いた発生工学的手法によりKAB-2チャネルのノックアウトマウスの作成を試みている。

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• 心筋細胞ATP依存性カリウムチャネルの受容体および機械刺激依存性制御とその異常

  倉智 嘉久, 高橋 尚彦, 山田 充彦, 内匠 透, 堀尾 嘉幸

  日本学術振興会, 科学研究費助成事業 重点領域研究, 重点領域研究, 大阪大学, 1994 - 1994

  ラットの心臓からATP依存性カリウムチャネル(K_チャネル)がクローニングされ(rcK_-1,Ashford et al.Nature370,456-459,1994)このチャネルの一次構造が明らかになった。研究代表者らはこのクローンを譲り受け,ヒト腎臓由来HEK293細胞にトランスフェクションし、パッチクランプ法を用い電気生理学的にこのチェネルの機能を解析した。これによって,このチャネルは心筋細胞から直接記録したK_チャネルと単一チャネルコンダクタンスの大きさや開閉のキネティクスなどはほぼ同一であるが,細胞内のATPに対する感受性が一定でないことや,グリベンクラミドなどのスルフォニルウレア剤によって抑制されない,などの相違点を有することが明らかになった。この相違の一つの機序として,このチャネルが相同性を有する異なるサブユニットの重合によって機能単位となることが示唆されたので,研究代表者らはrcK_-1をプローブにしてマウス脳のcDNAライブラリーをスクリーニングし,rcK_-1と約80%の相同性を有するチャネルおよびそのsplicing variantを単離することに成功した。現在これらの複数のクローンを組み合わせて発現させ,このチャネルがいかなるサブユニット構造をとって機能しているかを解析している。さらにキメラクローンや点変異導入クローンを作製し,種々のKチャネル開口薬やnucleotide diphosphateによるK_チャネル活性化の分子機構について実験を行っている。

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• 三量体G蛋白質による心筋細胞カリウムチャネル制御の分子機構

  倉智 嘉久, 高橋 尚彦, 山田 充彦, 内匠 透, 堀尾 嘉幸

  日本学術振興会, 科学研究費助成事業 重点領域研究, 重点領域研究, 大阪大学, 1994 - 1994

  本年度は、マウスのムスカリン性カリウム(K_)チャネルをクローニングし、このチャネルがG蛋白質βγサブユニット(G_<βγ>)と直接的に物理的結合を介して連関することを証明した。まず、ラット心臓から獲られたK_チャネルクローン(GIRK1)の一部をプローブとしてマウ脳をcDNAライブラリーをスクリーニングし、GIRK1と99%のアミノ酸配列相同性を有するクローン(MB-GIRK1)を得た。このクローンは、Xenopus oocyteでG蛋白質によって活性化される内向き整流カリウム電流を発現するので、マウスのK_チャネルクローンであると考えられた。MB-GIRK1はG蛋白質調節を受けない他の内向き整流カリウムチャネルのクローン(IRK1,2,3)と比較して♯アミノ酸長いC末を有する。そこでこの部位がG蛋白質との連関に関与するのではないかと考え、大腸菌を用いてこの配列のペプチドをGlutathione S-transferase融合蛋白質として形成し、G蛋白質αサブユニット(G_α)またはG_<βγ>との物理的連関の可能性を検討した。その結果、この融合蛋白質はG_<βγ>と結合するが活性型または非活性型のG_αとは結合しないこと、非活性型のG_αはG_<βγ>と融合蛋白質の結合を阻害するが活性型のG_αはこの効果を有さないことがin vitroで証明された。さらにインサイドアウトパッチ内のモルモットの心房筋細胞のK_チャネルは、パッチ膜の細胞内側から投与された10nMのG_<βγ>によって迅速かつ最大に活性化されるが、融合蛋白質と前処理したG_<βγ>と同濃度でこのチャネルを緩徐にかつ部分的に活性化したのみであった。したがって、MB-GIRKIはそのC末とG_<βγ>との物理的結合を介して直接G蛋白質と連関することが解かった。

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• NOによる中枢神経細胞内向き整流カリウムチャネルの抑制性修飾とその分子機構

  倉智 嘉久, 高橋 尚彦, 山田 充彦, 内匠 透, 堀尾 嘉幸

  日本学術振興会, 科学研究費助成事業 重点領域研究, 重点領域研究, 大阪大学, 1994 - 1994

  研究代表者らは,自らがマウス脳から単離した3種の内向き整流特性カリウムチャネル(IRK1,IRK2,IRK3)をアフリカツメガエルの卵母細胞に発現させ,細胞内サイクリックGMP(cGMP)およびサイクリックAMP(cAMP)の効果を検討した。この3つのチャネル電流はいずれも高濃度(〜2mM)の膜透過性cGMPおよびcAMPによって電位および時間依存性に抑制され,この効果は種々のカイネース阻害剤に影響されないことからリン酸化を介するものではないことが明らかになった。さらにIRKIとマウス脳由来のpoly(A^+)RNAを同時に発現させた卵母細胞では,イソプロテレノールがIRKI電流を抑制した。しかしこれらの抑制効果は約3割のツメガエルにみられるのみであり,また細胞外から与えたニトロプルシドはこのチャネル電流を抑制しなかった。さらに膜非透過性のcGMPおよびcGMPも弱い抑制効果を示した。以上の結果からcGMP(cAMP)によるIRK抑制効果は,細胞内からだけでなく細胞外からの経路もあり,また安定した抑制を示すには何らかの条件が必要であることが示唆された。この抑制経路および抑制のための至適条件を同定するために現在パッチクランプ法を用いた実験を行っている。さらに脳のスライス標本を用い,スライスパッチ法によりIRK電流の同定および膜透過性cGMP(cAMP)の効果を検討中である。

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• カリウムチャンネル活性を有する新しい膜蛋白(Isk)の構造と機能に関する研究

  内匠 透

  日本学術振興会, 科学研究費助成事業 奨励研究(特別研究員), 奨励研究(特別研究員), 京都大学, 1990 - 1990