Research activity information

• Sep. 2024 Japanese Society of Animal Science, The 2024 Award for Excellence in Reviewing for Animal Science Journal in 2023, Animal Science Journal Reviewers Award

  HARAYAMA Hiroshi


• Apr. 2021 Japanese Society of Animal Science, The 2021 Award for Excellence in Reviewing for Animal Science Journal in 2020, Animal Science Journal Reviewers Award

  HARAYAMA Hiroshi


• Mar. 2019 Japanese Society of Animal Science, The 2019 Award for Excellence in Reviewing for Animal Science Journal in 2018, Animal Science Journal Reviewers Award

  HARAYAMA Hiroshi

  Official journal


• Jul. 2017 JSPS, 日本学術振興会（JSPS）平成28年度 特別研究員等審査会専門委員（書面担当）の表彰, 日本学術振興会（JSPS）平成28年度 特別研究員等審査会専門委員（書面担当）の表彰

  HARAYAMA HIROSHI

  書面審査における有意義な審査意見を付した 専門委員として選ばれたため。

  Others


• Sep. 2012 The Society for Reproduction and Development (SRD), The 2012 SRD Outstanding Research Award, Study on intracellular cAMP signaling in mammalian spermatozoa

  HARAYAMA HIROSHI

  繁殖生物学に関する基盤研究、応用研究等に顕著な功績があった。

  Japan society


• Aug. 2006 関西畜産学会, 第56回関西畜産学会優秀発表賞（第1位）, ブタ精子の頭部におけるcAMP依存性細胞内カルシウム濃度上昇に及ぼす炭酸水素ナトリウムおよびPKA阻害剤の影響

  TATE Shunsuke, SHIDARA Osamu, HARAYAMA Hiroshi

  Japan society


• Apr. 2006 American Society of Andrology, 2006 Lalor Foundation Award, Role of protein kinase C in the cyclic adenosine 3',5'-monophosphate-dependent hyperactivation of boar spermatozoa

  HARAYAMA Hiroshi

  International society

• Large cyclic AMP-dependent increases in tyrosine-phosphorylated proteins are not absolutely necessary for the induction of full-type hyperactivation and penetration into eggs in ejaculated boar spermatozoa in vitro

  UESHIBA Ami, NAKAO Riko, MORIKAWA-TOMITA Riho, KYOGOKU Hirohisa, HARAYAMA Hiroshi

  2026, Journal of Reproduction and Development, 72(2) (2), 55 - 64, English

  [Refereed]

  Scientific journal


• Spermatic RXFP2 expression levels and seminal INSL3 concentrations among beef bull ejaculates with different levels of sperm morphological normality

  WIMALARATHNE Hewage Dilhan Anuradha, ARASHI Kenta, IWAKI Fumiyuki, SAKASE Mitsuhiro, DURITAHALA, HARAYAMA Hiroshi, KAWATE Noritoshi

  Feb. 2025, Journal of Reproduction and Development, 71(1) (1), 35 - 40, English

  [Refereed]

  Scientific journal


• Comparative characteristics between calyculin A-induced and thimerosal-induced hyperactivation of cryopreserved bovine spermatozoa

  MIYAMOTO Natsuko, OHYA Akihiro, DURITAHALA, SAKASE Mitsuhiro, HARAYAMA Hiroshi

  Jun. 2023, Journal of Reproduction and Development, 69(3) (3), 170 - 177, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Polyvinyl alcohol, but not bovine serum albumin, promotes the induction of full-type hyperactivation in boar cyclic AMP analog-treated spermatozoa

  SAI Seiken, HARAYAMA Hiroshi

  Nov. 2022, Animal Science JOurnal, 93, e13777, English

  [Refereed]

  Scientific journal


• Involvement of Ca²⁺-ATPase in suppressing the appearance of bovine helically motile spermatozoa with intense force prior to cryopreservation

  DURITAHALA, SAKASE Mitsuhiro, HARAYAMA Hiroshi

  Jun. 2022, Journal of Reproduction and Development, 68(3) (3), 181 - 189, English

  [Refereed]

  Scientific journal


• Calmodulin is involved in the occurrence of extracellular Ca²⁺-dependent full-type hyperactivation in boar ejaculated spermatozoa incubated with cyclic AMP analogs

  WADA Atsushi, HARAYAMA Hiroshi

  Apr. 2021, Animal Science Journal, 92(e13552) (e13552), English

  [Refereed]

  Scientific journal


• Effects of digoxin on full-type hyperactivation in bovine ejaculated spermatozoa with relatively lower survivability for incubation with stimulators of cAMP signaling cascades

  SAHA Soma Rani, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  Sep. 2020, Theriogenology, 154, 100 - 109, English

  [Refereed]

  Scientific journal


• Characteristics of bull sperm acrosome associated 1 proteins

  MINAMI Kenta, ARAI-ASO Miyuki M, OGURA-KODAMA Yukari, YAMADA Ayano, KISHIDA Kazumi, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  Aug. 2020, Animal Reproduction Science, 218(106479) (106479), English

  [Refereed]

  Scientific journal


• Identification of isoforms of calyculin A-sensitive protein phosphatases which suppress full-type hyperactivation in bull ejaculated spermatozoa

  ARAI Yuka, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  Apr. 2019, Theriogenology, 129, 46 - 53, English

  [Refereed]

  Scientific journal


• Flagellar hyperactivation of bull and boar spermatozoa [Invited review article]

  HARAYAMA Hiroshi

  Oct. 2018, Reproductive Medicine and Biology, 17(4) (4), 442 - 448, English

  [Refereed][Invited]


• Reconsideration of the evaluation criteria for bull ejaculated sperm motility in the context of rotation

  YAMADA Ayano, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  Oct. 2018, Journal of Reproduction and Development, 64(5) (5), 377 - 384, English

  [Refereed]

  Scientific journal


• Characterization of extracellular Ca²⁺-dependent full-type hyperactivation in ejaculated boar spermatozoa preincubated with a cAMP analog

  Nagisa Otsuka, Hiroshi Harayama

  Nov. 2017, MOLECULAR REPRODUCTION AND DEVELOPMENT, 84(11) (11), 1203 - 1217, English

  [Refereed]

  Scientific journal


• 家畜精子鞭毛の超活性化運動に関する研究の現状 [招待総説論文]

  原山 洋, 水野 洋平

  Sep. 2017, 日本胚移植学雑誌, 39(3) (3), 159 - 167, Japanese

  [Refereed][Invited]

  Scientific journal


• Protein biomarkers for male artificial insemination subfertility in bovine spermatozoa [Invited review article]

  HARAYAMA Hiroshi, MINAMI Kenta, KISHIDA Kazumi, NODA Taichi

  Apr. 2017, Reproductive Medicine and Biology, 16(2) (2), 89 - 98

  [Refereed][Invited]


• Variation among individual bulls in the distribution of acrosomal tyrosine-phosphorylated proteins in epididymal and ejaculated spermatozoa

  Miyuki M. Arai, Kenta Minami, Yukari Ogura, Nagisa Otsuka, Shohei Hama, Hiroshi Harayama, Mitsuhiro Sakase, Moriyuki Fukushima

  Corresponding, CSIRO, 2017, Reproduction, Fertility and Development, 29(7) (7), 1297 - 1305, English

  [Refereed]

  Scientific journal


• Changes of IZUMOI in bull spermatozoa during the maturation, acrosome reaction, and cryopreservation

  Masaki Fukuda, Mitsuhiro Sakase, Moriyuki Fukushima, Hiroshi Harayama

  Dec. 2016, THERIOGENOLOGY, 86(9) (9), 2179 - 2188, English

  [Refereed]

  Scientific journal


• Individual differences in the distribution of sperm acrosome-associated 1 proteins among male patients of infertile couples; their possible impact on outcomes of conventional in vitro fertilization

  K. Kishida, H. Harayama, F. Kimura, T. Murakami

  Oct. 2016, ZYGOTE, 24(5) (5), 654 - 661, English

  [Refereed]

  Scientific journal


• Changes in the distribution and molecular mass of boar sperm acrosome-associated 1 proteins during the acrosome reaction; their validity as indicators for occurrence of the true acrosome reaction

  Yukari Ogura, Yuki Takagishi, Hiroshi Harayama

  Sep. 2016, ANIMAL REPRODUCTION SCIENCE, 172(1) (1), 94 - 104, English

  [Refereed]

  Scientific journal


• Involvement of cAMP-dependent unique signaling cascades in the decrease of serine/threonine-phosphorylated proteins in boar sperm head

  Ayane Isono, Shunsuke Tate, Kazumi Nakamura-Mori, Taichi Noda, Sho Ishikawa, Hiroshi Harayama

  Apr. 2016, THERIOGENOLOGY, 85(6) (6), 1152 - 1160, English

  [Refereed]

  Scientific journal


• Effects of acrosomal conditions of frozen-thawed spermatozoa on the results of artificial insemination in Japanese Black cattle

  Kazumi Kishida, Mitsuhiro Sakase, Kenta Minami, Miyuki M. Arai, Reiko Syoji, Namiko Kohama, Takayuki Akiyama, Akio Oka, Hiroshi Harayama, Moriyuki Fukushima

  Corresponding, Dec. 2015, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 61(6) (6), 519 - 524, English

  [Refereed]

  Scientific journal


• Roles of extracellular Ca²⁺ in the occurrence of full-type hyperactivation in boar ejaculated spermatozoa pre-incubated to induce the cAMP-triggered events

  A. Kojima, Y. Matsushita, Y. Ogura, S. Ishikawa, T. Noda, T. Murase, H. Harayama

  Mar. 2015, ANDROLOGY, 3(2) (2), 321 - 331, English

  [Refereed]

  Scientific journal


• Distinct Segment-Specific Functions of Calyculin A-Sensitive Protein Phosphatases in the Regulation of cAMP-Triggered Events in Ejaculated Bull Spermatozoa

  Yohei Mizuno, Ayane Isono, Aya Kojima, Miyuki M. Arai, Taichi Noda, Mitsuhiro Sakase, Moriyuki Fukushima, Hiroshi Harayama

  Mar. 2015, MOLECULAR REPRODUCTION AND DEVELOPMENT, 82(3) (3), 232 - 250, English

  [Refereed]

  Scientific journal


• Expression patterns of the activator type of cAMP-responsive element modulator in testicular germ cells of Japanese Black bulls

  Taichi Noda, Kenta Minami, Aya Kojima, Yohei Mizuno, Ayane Isono, Mitsuhiro Sakase, Moriyuki Fukushima, Hiroshi Harayama

  Last, May 2014, THERIOGENOLOGY, 81(8) (8), 1012 - 1020, English

  [Refereed]

  Scientific journal


• Roles of intracellular cyclic AMP signal transduction in the capacitation and subsequent hyperactivation of mouse and boar spermatozoa [Invited review article]

  HARAYAMA Hiroshi

  Oct. 2013, Journal of Reproduction and Development, 59(5) (5), 421 - 430, English

  [Refereed][Invited]

  Scientific journal


• Novel Approach for the Detection of the Vestiges of Testicular mRNA Splicing Errors in Mature Spermatozoa of Japanese Black Bulls

  Taichi Noda, Mitsuhiro Sakase, Moriyuki Fukushima, Hiroshi Harayama

  Last, Feb. 2013, PLOS ONE, 8(2) (2), e57296, English

  [Refereed]

  Scientific journal


• Relationship between cyclic AMP-dependent protein tyrosine phosphorylation and extracellular calcium during hyperactivation of boar spermatozoa

  Hiroshi Harayama, Taichi Noda, Shou Ishikawa, Osamu Shidara

  Oct. 2012, MOLECULAR REPRODUCTION AND DEVELOPMENT, 79(10) (10), 727 - 739, English

  [Refereed]

  Scientific journal


• Detection of the activator cAMP responsive element modulator (CREM) isoform ortholog proteins in porcine spermatids and sperm

  T. Noda, O. Shidara, H. Harayama

  Last, Apr. 2012, THERIOGENOLOGY, 77(7) (7), 1360 - 1368, English

  [Refereed]

  Scientific journal


• Effect of H89 on the meiotic resumption of pig oocytes

  Ilse Silvia Cayo-Colca, Hiroshi Harayama, Takashi Miyano

  John Wiley and Sons Ltd, 2011, Reproductive Medicine and Biology, 10(2) (2), 89 - 96, English

  [Refereed]

  Scientific journal


• Relationship of Protein Tyrosine Phosphorylation State With Tolerance to Frozen Storage and the Potential to Undergo Cyclic AMP-Dependent Hyperactivation in the Spermatozoa of Japanese Black Bulls

  Hiroshi Harayama, Kazuhiro Nishijima, Tetsuma Murase, Mitsuhiro Sakase, Moriyuki Fukushima

  Oct. 2010, MOLECULAR REPRODUCTION AND DEVELOPMENT, 77(10) (10), 910 - 921, English

  [Refereed]

  Scientific journal


• Evidence of the Existence of Adenylyl Cyclase 10 (ADCY10) Ortholog Proteins in the Heads and Connecting Pieces of Boar Spermatozoa

  Shunsuke Tate, Kazumi Nakamura, Chihiro Suzuki, Taichi Noda, Jibak Lee, Hiroshi Harayama

  Apr. 2010, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 56(2) (2), 271 - 278, English

  [Refereed]

  Scientific journal


• Hyperactivated Motility of Frozen-Thawed Spermatozoa from Fertile and Subfertile Japanese Black Bulls Induced by Cyclic Adenosine 3 ',5 '-Monophosphate Analogue, cBiMPS

  Tetsuma Murase, Ismail El-Kon, Hiroshi Harayama, Koushi Mukoujima, Masaki Takasu, Kenji Sakai

  Feb. 2010, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 56(1) (1), 36 - 40, English

  [Refereed]

  Scientific journal


• When does the sex ratio of offspring of the paternal 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) exposure decrease: In the spermatozoa stage or at fertilization?

  Kana Ishihara, Seiichiroh Ohsako, Ken Tasaka, Hiroshi Harayama, Masashi Miyake, Katsuhiko Warita, Takashi Tanida, Tomoko Mitsuhashi, Takashi Nanmori, Yoshiaki Tabuchi, Toshifumi Yokoyama, Hiroshi Kitagawa, Nobuhiko Hoshi

  Jan. 2010, REPRODUCTIVE TOXICOLOGY, 29(1) (1), 68 - 73, English

  [Refereed]

  Scientific journal


• Calyculin A-Sensitive Protein Phosphatases are Involved in Maintenance of Progressive Movement in Mouse Spermatozoa In Vitro by Suppression of Autophosphorylation of Protein Kinase A

  Namiko Goto, Hiroshi Harayama

  Jun. 2009, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 55(3) (3), 327 - 334, English

  [Refereed]

  Scientific journal


• A 32-kDa Tyrosine-phosphorylated Protein Shows a Protease-dependent Increase in Dead Boar Spermatozoa

  Tomohito Tabuchi, Osamu Shidara, Hiroshi Harayama

  Dec. 2008, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 54(6) (6), 502 - 507, English

  [Refereed]

  Scientific journal


• Changes of PKA and PDK1 in the principal piece of boar spermatozoa treated with a cell-permeable cAMP analog to induce flagellar hyperactivation

  Hiroshi Harayama, Kazumi Nakamura

  Sep. 2008, MOLECULAR REPRODUCTION AND DEVELOPMENT, 75(9) (9), 1396 - 1407, English

  [Refereed]

  Scientific journal


• Effects of protein phosphatase inhibitor calyculin a on the postacrosomal protein serine/threonine phosphorylation state and acrosome reaction in bear spermatozoa incubated with a cAMP analog

  Jun Adachi, Shunsuke Tate, Masashi Miyake, Hiroshi Harayama

  Jun. 2008, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 54(3) (3), 171 - 176, English

  [Refereed]

  Scientific journal


• Premature Capacitation of Frozen-Thawed Spermatozoa from Subfertile Japanese Black Cattle

  KURODA Ken, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  Nov. 2007, J. Reprod. Dev., 53(5), pp.1079-1086, English

  [Refereed]

  Scientific journal


• Evidence for existence of cAMP-Epac signaling in the heads of mouse epididymal spermatozoa

  Reiko Amano, Jibak Lee, Namiko Goto, Hiroshi Harayama

  Feb. 2007, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 53(1) (1), 127 - 133, English

  [Refereed]

  Scientific journal


• A cyclic adenosine 3',5'-monophosphate-dependent protein kinase C activation is involved in the hyperactivation of boar spermatozoa.

  HARAYAMA Hiroshi, MIYAKE Masashi

  Sep. 2006, Mol. Reprod. Dev., 73(9), pp.1169-1178, 1169 - 1178, English

  [Refereed]

  Scientific journal


• The localization of LAP2 beta during pronuclear formation in bovine oocytes after fertilization or activation

  M Isaji, H Iwata, H Harayama, M Miyake

  May 2006, ZYGOTE, 14(2) (2), 157 - 167, English

  [Refereed]

  Scientific journal


• A cyclic adenosine 3 ',5 '-monophosphate stimulates phospholipase C gamma 1-calcium signaling via the activation of tyrosine kinase in boar spermatozoa

  H Harayama, T Murase, M Miyake

  Nov. 2005, JOURNAL OF ANDROLOGY, 26(6) (6), 732 - 740, English

  [Refereed]

  Scientific journal


• Exogenous hyaluronic acid enhances porcine parthenogenetic embryo development in vitro possibly mediated by CD44

  Toyokaw K, Harayama H, Miyake M

  Jan. 2005, Theriogenology, 64(1) (1), 378 - 392, English

  [Refereed]

  Scientific journal


• A cyclic adenosine 3 ',5 '-monophosphate-induced tyrosine phosphorylation of Syk protein tyrosine kinase in the flagella of boar spermatozoa

  H Harayama, M Muroga, M Miyake

  Dec. 2004, MOLECULAR REPRODUCTION AND DEVELOPMENT, 69(4) (4), 436 - 447, English

  [Refereed]

  Scientific journal


• A unique mechanism for cyclic adenosine 3 ',5 '-monophosphate-induced increase of 32-kDa tyrosine-phosphorylated protein in boar spermatozoa

  H Harayama, K Sasaki, M Miyake

  Oct. 2004, MOLECULAR REPRODUCTION AND DEVELOPMENT, 69(2) (2), 194 - 204, English

  [Refereed]

  Scientific journal


• The localization of LAP2 beta in bovine oocytes after in vitro activation and fertilization

  M Isaji, H Iwata, H Harayama, M Miyake

  Feb. 2004, ZYGOTE, 12(1) (1), 81 - 93, English

  [Refereed]

  Scientific journal


• Viability and protein phosphorylation patterns of boar spermatozoa agglutinated by treatment with a cell-permeable cyclic adenosine 3 ',5 '-monophosphate analog

  H Harayama

  Nov. 2003, JOURNAL OF ANDROLOGY, 24(6) (6), 831 - 842, English

  [Refereed]

  Scientific journal


• Effects of season on concentrations of seminal plasma acidic proteins from Japanese Black cattle

  西本 尚美, 中村 弘毅, 岩木 史之, 岡 章生, 福島 護之, 太田垣 進, 原山 洋

  Sep. 2003, Japanese Journal of Embryo Transfer, 25,pp.101-107(3) (3), 101 - 107, Japanese

  [Refereed]

  Scientific journal


• Stage-specific effects of the osmolarity of a culture medium on the development of parthenogenetic diploids in the pig

  VT Nguyen, S Kure-bayashi, H Harayama, T Nagai, M Miyake

  Feb. 2003, THERIOGENOLOGY, 59(3-4) (3-4), 719 - 734, English

  [Refereed]

  Scientific journal


• Involvement of cytoplasmic free calcium in boar sperm: head-to-head agglutination induced by a cell-permeable cyclic adenosine monophosphate analog

  HARAYAMA Hiroshi, OKADA K, MIYAKE Masashi

  Feb. 2003, J. Androl., 24,pp.91-99, 91 - 99, English

  [Refereed]

  Scientific journal


• Localization of Lap2B in oocytes and embryos during early development in cattle

  Isaji M, Iwata H, Ohota M, Harayama H, Miyake M

  LAP2βはラミンの裏打ち蛋白であるがその機能については不明な点が多い。ウシ胚においてこのLAP2βの局在と発現について観察した。LAP2βの発現や局在は体外受精卵や単為発生胚で差がなく発生が進むにつれて前核期よりも発現が強くなることを示した。

  Jan. 2003, Theriogenology 第59巻 474 (abstr), English

  [Refereed]

  Scientific journal


• Capacitation-like alterations in cooled boar spermatozoa: assessment by the chlortetracycline staining assay and immunodetection of tyrosine-phosphorylated sperm proteins

  M Kaneto, H Harayama, M Miyake, S Kato

  Corresponding, Oct. 2002, ANIMAL REPRODUCTION SCIENCE, 73(3-4) (3-4), 197 - 209, English

  [Refereed]

  Scientific journal


• Relationship between bicarbonate and cyclic nucleotide in the promoting effects on head-to-head agglutination in boar spermatozoa

  H Harayama, S Kato

  Jun. 2002, ASIAN JOURNAL OF ANDROLOGY, 4(2) (2), 87 - 96, English

  [Refereed]

  Scientific journal


• Characteristics of preimplantational development of porcine parthenogenetic diploids relative to the existence of amino acids in vitro

  Van Thuan N, Hiroshi HARAYAMA, Masashi MIYAKE

  Jun. 2002, Biol Reprod., 67(6), pp. 1688-1698, 1688 - 1698, English

  [Refereed]

  Scientific journal


• The development of porcine parthenogenetic diploid oocytes with homogeneous genomic components in vitro

  NV Thuan, H Harayama, M Miyake

  Apr. 2002, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 48(2) (2), 157 - 166, English

  [Refereed]

  Scientific journal


• Detection of a fertility-associated protein osteopontin in seminal plasma from Japanese Black cattle

  FURUTSUKA Masayuki, HARAYAMA Hiroshi, MIYAKE Masashi, FUKUSHIMA Moriyuki, OHTAGAKI Susumu, KATO Seishiro

  Corresponding, Jan. 2002, Japanese Journal of Embryo Transfer, 24(1), pp.1-8(1) (1), 1 - 8, Japanese

  [Refereed]

  Scientific journal


• Factors Regulating Changes of Head-to-Head Agglutinability in Boar Spermatozoa during Epididymal Transit and Capacitation in Vitro - Review -

  Hiroshi Harayama, Seishiro Kato

  2001, Asian-Australasian Journal of Animal Sciences, 14(8) (8), 1196 - 1202

  [Refereed][Invited]


• Biochemical characterization of sialoprotein "anti-agglutinin" purified from boar epididymal and seminal plasma

  H Harayama, PC Liao, DA Gage, M Miyake, S Kato, RH Hammerstedt

  Jan. 2000, MOLECULAR REPRODUCTION AND DEVELOPMENT, 55(1) (1), 96 - 103, English

  [Refereed]

  Scientific journal


• Role of cyclic adenosine 3 ',5 '-monophosphate and serum albumin in head-to-head agglutination of boar spermatozoa

  H Harayama, M Miyake, S Kato

  2000, REPRODUCTION FERTILITY AND DEVELOPMENT, 12(5-6) (5-6), 307 - 318, English

  [Refereed]

  Scientific journal


• Immunolocalization of anti-agglutinin for spermatozoa in boars

  H Harayama, M Miyake, S Kato

  Mar. 1999, MOLECULAR REPRODUCTION AND DEVELOPMENT, 52(3) (3), 269 - 276, English

  [Refereed]

  Scientific journal


• Changes in epididymal protein anti-agglutinin on ejaculated boar spermatozoa during capacitation in vitro

  H Harayama, SF Magargee, E Kunze, O Shidara, E Iwamoto, S Arikawa, M Miyake, S Kato, RH Hammerstedt

  1999, REPRODUCTION FERTILITY AND DEVELOPMENT, 11(4-5) (4-5), 193 - 199, English

  [Refereed]

  Scientific journal


• 冷却処理後の梅山豚射出精子の運動性および先体の形態に及ぼす精漿成分の影響

  原山 洋, 今野 彰, 三宅 正史, 加藤 征史郎

  Lead, Aug. 1998, 日本畜産学会報, 69(8) (8), 720 - 727, Japanese

  [Refereed]

  Scientific journal


• Effects of seminal plasma components on motility and acrosomal integrity of Meishan boar spermatozoa after cooling treatments

  Hiroshi HARAYAMA, Akira IMANO, Seishiro KATO

  Aug. 1998, Anim. Sci. Technol. (Jpn), 69(8), pp. 720-727(8) (8), 720 - 727, English

  [Refereed]

  Scientific journal


• Bone histomorphology in tibial diaphysis of growing chicks administered excessive vitamin A

  Yasuyuki KANNAN, Hiroshi HARAYAMA, Seishiro KATO

  The effect of hypervitaminosis A on bone histomorphology was studied in the tibiae of young growing chicks. Male White Leghorn chicks were orally administered excessive vitamin A (300 or 600IU per g body weight per day) for 10 consecutive days. For histological examination, middle diaphyses were excised and cross sections were stained with toluidine blue. Morphological differences were examined by a computerlized histomorphometry. Radiographic and histological observations indicated the hypervitaminotic chicks to have short tibial length, narrow diaphyseal width and abnormal bone morphology characterized by flattened spindle-shaped osteoblasts. Histomorphometric analysis indicated excessive vitamin A intake to cause severe retardation of bone formation. Cortical bone thickness, measured at middle diaphysis, was less in hypervitaminotic than control chicks receiving the vehicle alone. Osteoblast area and number significantly decreased in the hypervitaminotic chicks. Changes in these osteoblast parameters are associated with marked decrease in bone volume, expressed as percentage of mineralized bone matrix area. Excessive vitamin A intake did not affect all histomorphometric indices of bone resorption. Hypervitaminosis A would thus appear to decrease osteoblastic activity and inhibit bone formation in chick tibia.

  Japan Poultry Science Association, Mar. 1998, Japanese Poultry Science, 35(2), pp. 108-116(2) (2), 108 - 116, English

  [Refereed]

  Scientific journal


• Lectin-binding characteristics of extracts from epididymal boar spermatozoa

  Hiroshi Harayama, Shin-Ya Watanabe, Hiroshi Masuda, Yasuyuki Kannan, Masashi Miyake, Seishiro Kato

  The Japanese Society of Animal Reproduction (JSAR), 1998, Journal of Reproduction and Development, 44(1) (1), 21 - 27, English

  [Refereed]

  Scientific journal


• Effects of calcium and bicarbonate on head-to-head agglutination in ejaculated boar spermatozoa

  H Harayama, M Miyake, O Shidara, E Iwamoto, S Kato

  1998, REPRODUCTION FERTILITY AND DEVELOPMENT, 10(5) (5), 445 - 450, English

  [Refereed]

  Scientific journal


• Effects of dietary calcium levels on the histomorphology of proximal tibia in vitamin D-deficient chicks

  Yasuyuki KANNAN, Hiroshi HARAYAMA, Seishiro KATO

  Mar. 1997, Japanese Poultry Science, 34(2), pp. 124-131, 107 - 116, English

  [Refereed]

  Scientific journal


• Electrophoretic characterization of boar epididymal antiagglutinin

  H Harayama, S Kato, RH Hammerstedt

  Aug. 1996, BIOLOGY OF REPRODUCTION, 55(2) (2), 325 - 332, English

  [Refereed]

  Scientific journal


• Timing of shedding and disintegration of cytoplasmic droplets from boar and goat spermatozoa

  Seishiro Kato, Toshitaka Shibukawa, Hiroshi Harayama, Yasuyuki Kannan

  The Japanese Society of Animal Reproduction (JSAR), 1996, Journal of Reproduction and Development, 42(4) (4), 237 - 241, English

  [Refereed]

  Scientific journal


• Fructose stimulates shedding of cytoplasmic droplets from epididymal boar spermatozoa

  H Harayama, M Miyake, T Shibukawa, Y Kannan, S Kato

  1996, REPRODUCTION FERTILITY AND DEVELOPMENT, 8(7) (7), 1039 - 1043, English

  [Refereed]

  Scientific journal


• Detection of 25-kDa Anti-Agglutinin in Epididymal Plasma and Spermatozoa Collected from Various Regions of Boar Epididymis

  Hiroshi Harayama, Takashi Miyano, Seishiro Kato, Hiroshi Masuda, Masashi Miyake

  1995, Journal of Reproduction and Development, 41(2) (2), 113 - 121, English

  [Refereed]

  Scientific journal


• IDENTIFICATION OF ANTI-AGGLUTININ FOR SPERMATOZOA IN EPIDIDYMAL BOAR PLASMA

  H HARAYAMA, T MIYANO, M MIYAKE, H KUSUNOKI, S KATO

  Apr. 1994, MOLECULAR REPRODUCTION AND DEVELOPMENT, 37(4) (4), 436 - 445, English

  [Refereed]

  Scientific journal


• CAPACITY OF RETE TESTICULAR AND CAUDA EPIDIDYMAL BOAR SPERMATOZOA TO UNDERGO THE ACROSOME REACTION AND SUBSEQUENT FUSION WITH EGG PLASMA-MEMBRANE

  H HARAYAMA, H KUSUNOKI, S KATO

  May 1993, MOLECULAR REPRODUCTION AND DEVELOPMENT, 35(1) (1), 62 - 68, English

  [Refereed]

  Scientific journal


• Characteristics of the ejaculates from Meishan boars at various ages

  Hiroshi HARAYAMA, Seishiro KATO

  The age at which ejaculatory ability is achieved and age-related changes in the characteristics of ejaculates were examined in Meishan boars. All of the 12 boars examined showed detachment of the penis from the prepuce by 64 days of age. The first ejaculate could be collected at 73 to 78 days of age. The volume of the liquid portion, the weight of the gelatinous material, the total number of spermatozoa per ejaculate and the sperm concentration increased significantly until 11, 10, 9 and 5 months of age, respectively. Percentages of progressively motile spermatozoa and morphologically abnormal spermatozoa in the ejaculates altered greatly until 3 months of age, and thereafter stayed at a good level. Percentages of live spermatozoa with normal acrosomes after storage at 4°C for 5 and 10 days were higher in ejaculates collected from boars older than 9 months, and percentages of progressively motile spermatozoa and live spermatozoa with normal acrosomes after freeze-thawing in boars older than 8 months. These results suggest that Meishan boars acquire ejaculatory ability at approximately 2 months of age, but should not be used in service until later ages. It is also suggested and that ejaculates from boars younger than 9 months are less suitable for storage at 4°C-196°C.

  Japanese Society of Animal Science, Apr. 1993, Anim. Reprod. Technol. (Jpn.), 64(4), pp. 333-339(4) (4), 333 - 339, English

  [Refereed]

  Scientific journal


• Motility and Penetrability into Zona-Free Hamster Eggs of Boar Spermatozoa Collected from Various Regions of the Epididymis

  Hiroshi Harayama, Hiroshi Kusunoki, Seishiro Kato

  1993, Journal of Reproduction and Development, 39(1) (1), 41 - 45, English

  [Refereed]

  Scientific journal


• CAPACITY OF GOAT EPIDIDYMAL SPERMATOZOA TO UNDERGO THE ACROSOME REACTION AND SUBSEQUENT FUSION WITH THE EGG PLASMA-MEMBRANE

  H HARAYAMA, H KUSUNOKI, S KATO

  1993, REPRODUCTION FERTILITY AND DEVELOPMENT, 5(3) (3), 239 - 246, English

  [Refereed]

  Scientific journal


• Development of the accessory genital glands in Meishan boars

  Hiroshi HARAYAMA, Sunao KANDA, Seishiro KATO

  The developmental process of the seminal vesicle, corpus prostatae and bulbourethral gland was morphologically and histologically investigated in Meishan boars after birth. All of the glands remarkably increased in weight after 30-45 days of age. In the seminal vesicle and corpus prostatae at 1 day of age, the glandular tubules were characterized by an undeveloped epithelium and a narrow lumen. At 45-60 days of age, the terminal portions of the tubules of these glands were partially ramified and the lumen contained PAS-positive secretions. At 75-90 days of age, the portions presented a fully-developed structure. On the other hand, in the bulbourethral gland at 1 day of age, the terminal portions of the glandular tubules were already ramified and contained PAS-positive substances. At 30-45 days of age, they showed rapid growth and more ramifications and were filled with the secretions. After this age, their structural aspects presented no changes. These results indicate that the secrectory function of the seminal vesicle, corpus prostatae and bulbourethral gland in Meishan boars in well developed at 75-90, 75-90 and 30-45 days of age, respectively.

  Japanese Society of Animal Science, Nov. 1992, Anim. Sci. Technol. (Jpn.), 63(11), pp. 1115-1122(11) (11), 1115 - 1122, English

  [Refereed]

  Scientific journal


• INFLUENCE OF SEASON ON CHARACTERISTICS OF EPIDIDYMAL AND EJACULATED SEMEN IN MEISHAN BOARS

  H HARAYAMA, S KANDA, S KATO

  Sep. 1992, THERIOGENOLOGY, 38(3) (3), 491 - 500, English

  [Refereed]

  Scientific journal


• Seasonal changes of reproductive performance in Meishan boars

  Hiroshi HARAYAMA, Masashi MIYAKE, Seishiro KATO

  Lead, Aug. 1992, Proceedings of the International Symposium on Chinese Pig Breeds., pp. 162-165, English

  [Refereed]

  International conference proceedings


• In vitro storage of Meishan boar spermatozoa

  Seishiro KATO, Koichiro KANO, Hirioshi HARAYAMA, Hiroshi KUSUNOKI, Iwao NANJO

  Aug. 1992, Proceedings of the International Symposium on Chinese Pig Breeds, pp. 582-585, English

  [Refereed]

  International conference proceedings


• Changes in motility and morphology of spermatozoa during their transit through the epididymis in Meishan boars at various ages

  Hiroshi HARAYAMA, Seishiro KATO

  Changes in motility and morphology of spermatozoa during their transit through the epididymis were investigated in Meishan boars at various ages. At 75 days of age, only 26% of spermatozoa from the cauda epididymidis were progressively motile, 59% presented a morphological abnormality, mainly an abnormal head, and 62% had a cytoplasmic droplet at the proximal region of the middle piece. At 105-120 days of age, however, 89% of spermatozoa showed intensive, forward movement and 98% had a distal droplet after epididymal transit. At more than 90 days of age, the proportion of spermatozoa with an abnormal head significantly decreased during their transit through the caput epididymidis. These results indicate that epididymal functions associated with sperm maturation and selective exclusion of abnormal spermatozoa in Meishan boars are fully developed by approximately 120 days of age.

  Japanese Society of Animal Science, May 1992, Anim. Sci. Technol. (Jpn.), 63(5), pp. 462-467(5) (5), 462 - 467, English

  [Refereed]

  Scientific journal


• Development of the reproductive organs in Jinhua boars

  Hiroshi HARAYAMA, Akio Tsutsui, Seishiro KATO

  The developmental process of the reproductive organs was morphologically and histologically investigated in Jinhua boars after birth. The testis greatly developed after 45 days of age. Spermatozoa were first found in the seminiferous tubules at approximately 75 days of age, when the histological composition of the seminiferous epithelium improved to a mature type. Epididymis grew rapidly after 45 days of age, and revealed a well-developed structure of the ductal epithelium in any region by 60-75 days of age. On the other hand, the weight of each accessory genital gland increased rapidly after 60 days of age. In the seminal vesicle, corpus prostatae and blubourethral gland, the terminal portions were well ramified and filled with PAS-positive secretions by 75-90, 90 and 30-45 days of age, respectively. These results indicate that Jinhua boars reach puberty as early as 75 days of age. It also appears that development of the reproductive organs occurs at an earlier age in Jinhua boars than in European and American breeds.

  The Japanese Society of Swine Science, Feb. 1992, Jpn. J. Swine Sci, 29(3), pp. 174-183(3) (3), 174 - 182, Japanese

  [Refereed]

  Scientific journal


• Development of the epididymis in Meishan boars

  Hiroshi HARAYAMA, Iwao NANJO, Sunao KANDA, Seishiro KATO

  Feb. 1992, Asian-Aust. J. Anim. Sci., 5(1), pp. 165-171, English

  [Refereed]

  Scientific journal


• TESTICULAR DEVELOPMENT IN CHINESE MEISHAN BOARS

  H HARAYAMA, NANJO, I, S KANDA, S KATO

  Oct. 1991, THERIOGENOLOGY, 36(4) (4), 637 - 643, English

  [Refereed]

  Scientific journal


• Effect of imidazole in diluent on motility and acrosomal morphology of Meishan boar spermatozoa stored at 4 C

  Koichiro KANO, Hiroshi KUSUNOKI, Hiroshi HARAYAMA, Seishiro KATO

  Sep. 1991, Jpn. J. Anim. Reprod. Technol., 13(3), pp. 158-164, Japanese

  [Refereed]

  Scientific journal


• In vitro fertilization of in vitro and in vivo matured oocytes from prepubertal Meishan gilts

  Takashi MIYANO, Kiyoshi YOSHIKAWA, Seishiro KATO, Hiroshi HARAYAMA, Sunao KANDA

  Using prepubertal Meishan gilts, at 70-71 days of age, weighting 20-30 kg, we examined the details of the fertilization of in vitro and in vivo matured oocytes. At 39-40 h after hCG injection, follicles larger than 7mm in diameter were observed on the surface of ovaries(18.4±11.3, n=4). At 42-43h, all of the ovaries had recently ovulated follicles (6.5±3.7, n=4)in addition to follicles larger than 7mm in diameter. Immature oocytes were collected from hormone-nontreated gilts, and matured in a maturation medium. The cumuli surrounding follicular oocytes, recovered from the hormone-nontreated gilts, expanded after culture in a maturation medium. Follicular oocytes and ovulated eggs from hormone-treated gilts also had expanded cumuli. At 12h after insemination of the in vitro and in vivo matured follicular oocytes and of the ovulated eggs with the same sperm suspension, 72.3, 72.3% of oocytes and 84.4% of eggs reached the metaphase II, respectively. Formation of a male pronucleus (ei) was delayed in the oocytes matured in vitro, although the percentages of penetrated oocytes were not different among those three groups. The mean number of spermatozoa penetrating an ovulated egg was 2.1± 1.0, while the number was significantly higher in oocytes matured in vitro (3.6±2.8, p<0.05). These results show that mature oocytes and eggs recovered from prepubertal Meishan gilts as early as 70 days of age can be fertilized, we suggest that the Meishan gilt is a promising donor of oocytes and eggs for study of pig reproduction.

  Japanese Society of Animal Science, Nov. 1990, Jpn. J. Zootech. Sci., 61(11), pp. 1011-1016(11) (11), 1011 - 1016, English

  [Refereed]

  Scientific journal


• Retention of motility, viability and penetrating ability of acrosome-reacted goat spermatozoa

  Hiroshi KUSUNOKI, Morito SAKAUE, Hiroshi HARAYAMA, Seishiro KATO, Sunao KANDA

  The retention of the motility and viability of acrosome-reacted goat spermatozoa and their ability to penetrate zona-free hamster eggs was investigated. Effects of caffeine and imidazole on these parameters were also examined. Ejaculated goat spermatozoa were washed and preincubated in K-3 medium in sealed glass tubes at 39.5°C for 2h to induce the acrosome reaction. This was fo11owed by resuspension in BO medium BO medium with caffeine and BO medium with imidazole and incubated at 37°C for 5 h. During incubation, sperm motility and viability were assessed at one hour intervals. The acrosome-reacted cells after resuspension in BO medium or BO medium with imidazole and those incubated for 3h in these media were further incubated with zonafree hamster eggs at 37°C. During incubation with the eggs, time-related changes in egg penetration were examined. The following results were obtained: 1) The motility patterns of goat spermatozoa before and after the acrosome reaction were different. Acrosome-reacted goat spermatozoa exhibited 'whiplash' motility. 2) The motility and viability of goat cells after the acrosome reaction were remarkably lower than those before the reaction. 3) The ability of acrosome-reacted goat spermatozoa topenetrate zona-free hamster eggs was retained for as much as 3h. 4) The addition of imidazole to sperm suspension promoted the maintenance of motility and viability of acrosome-reacted goat spermatozoa and prolonged the retention of their ability to penetrate zona-free hamster eggs. The addition of caffeine was effective for the temporary activation of the motility but made difficult the maintenance of motility and viability.

  Japanese Society of Animal Science, Jul. 1990, Jpn. J. Zootech. Sci., 61(7), pp. 640-647(7) (7), 640 - 647, English

  [Refereed]

  Scientific journal


• Identification of acrosome-reacted goat spermatozoa by a trypan blue-Giemsa method

  Hiroshi KUSUNOKI, Morito SAKAUE, Hiroshi HARAYAMA, Seishiro KATO, SUNAO KANDA

  The purpose of this study was to develop a new technique for identifying acrosomereacted goat spermatozoa. Ejaculated spermatozoa were washed and incubated in an airtight glass tube at 39.5°C for 1, 2 and 3hrs for inducing the acrosome reaction, and washed and frozen-thawed three times. Sperm samples were incubated with trypan blue for 15min, smeared on a slide, fixed in ORTH solution for 45min, and finally stained in Giemsa solution (trypan blue-Giemsa method). Stained smears had spermatozoa with the following four staining patterns: a) unstained or light blue postacrosomal regions with reddish purple acrosomes (live sperm with normal acrosomes); b) unstained or light blue postacrosomal regions with unstained acrosomal regions or damaged acrosomes (live sperm without normal acrosomes); c) dark blue postacrosomal regions with dark reddish purple acrosomes (dead sperm with normal acrosomes); d) dark blue postacrosomal regions with dark blue acrosomal regions or damaged acrosomes (dead sperm without normal acrosomes). There was a significant positive partial correlation between the percentage of live sperm without normal acrosomes (category b) and that of zona-free hamster eggs penetrated by spermatozoa (r=0.4179, P〈0.01, n=44). Additionally, there was no difference between the results obtained by means of the present method and a triple-stain technique, while the staining procedure and solution preparation were much simpler in the former method than in the latter. These results indicate that the trypan blue-Giemsa method can be used to identify acrosome-reacted goat spermatozoa.

  Japanese Society of Animal Science, Mar. 1988, JPN. J. Zootech. Sci., 59(3), pp. 235-240(3) (3), 235 - 240, Japanese

  [Refereed]

  Scientific journal


• Storage of Meishan pig spermatozoa

  Hiroshi HARAYAMA, Seishiro KATO, Iwao NANJO, Sunao KANDA, Tetsushi TOKUMARU

  Lead, Jan. 1988, Jpn. J. Anim. Reprod. Technol., 10(1), pp. 10-14, Japanese

  [Refereed]

  Scientific journal


• Histological changes of testis and caput epididymis in the goat after cannulation of the rete testis

  Hiroshi HARAYAMA, Morito SAKAUE, Hiroshi KUSUNOKI, Seishiro KATO, Sunao KANDA

  雄ヤギ5頭を用い, 精巣網カテーテル装着手術後の精巣および精巣上体頭の組織構造を観察するとともに精巣静脈血中のテストステロン濃度を調べた。カテーテル装着手術後の精巣は, いずれも萎縮し, 正常な精子形成過程が観察される精細管は少なかった。また, ライディヒ細胞の形態や染色性には異常は認められなかった。一方, 精巣上体頭では, 精巣上体管が萎縮し, 主細胞の高さは無傷のものよりも有意に減少した。同様の組織構造の変化は, 精巣輸出管を切除した場合の精巣および精巣上体頭でも認められた。精巣静脈血中のテストステロ

  Lead, 神戸大学農学部, Jan. 1988, Sci. Rept. Fac. Agr. Kobe Univ., 18(1), pp. 125-132(1) (1), 125 - 132, Japanese

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Concetrations of steroid hormones in rete testicular plasma of the goat

  Morito SAKAUE, Osamu Doi, Hiroshi KUSUNOKI, Hiroshi HARAYAMA, Seishiro KATO, Sunao KANDA

  4頭の成熟雄ヤギ(日本在来種)から精巣網カテーテル法によって採取した精巣網液および頚静脈血を用い, ラジオイムノアッセイによりテストステロン, エストラジオール-17βおよびプロジェステロン濃度を測定した。精巣網漿液中のホルモンの平均濃度は, テストステロンで67.8ng/ml, エストラジオール-17βで43.9pg/ml, プロジェステロンでは517.1pg/mlであった。どの個体においても昼間(0900-2100)と夜間(2100-0900)に採取した試料のテストステロン濃度に有意な差は認められなか

  神戸大学農学部, Jan. 1988, Sci. Rept. Fac. Agr. Kobe Univ., 18, pp. 133-136(1) (1), 133 - 136, English

  Scientific journal

• 家畜精子での受精制御分子について

  HARAYAMA Hiroshi

  Oct. 2019, 日本アンドロロジー学会ニュースレター, 19, 3 - 4, Japanese

  [Invited]

  Others


• 特集 哺乳類の精子研究における新展開 －基礎と応用― 序

  HARAYAMA Hiroshi

  日本胚移植研究会, Sep. 2017, 日本胚移植学雑誌, 39(3) (3), 141, Japanese

  [Invited]

  Others


• Fertility Suppression of Feral Animal to Reconstitute Environmental Relationship with Human Society

  HARAYAMA Hiroshi, GOTO Namiko, TATE Shunsuke

  Apr. 2008, Annual Report of Interdisciplinary Research Institute of Environmental Sciences, 26, pp. 45-52, Japanese

  Others


• 精子鞭毛のハイパーアクチベーションを指標とする黒毛和種雄ウシの繁殖能力評価法の開発

  HARAYAMA Hiroshi

  Dec. 2007, 財）三島海雲記念財団平成18年度受贈者研究報告書, 44, pp. 19-23, Japanese

  Others


• Establishment of simplified assay for the reproductive performance of male Japanese Black cattle: additional application of sperm CTC staining technique

  Hiroshi HARAYAMA, Ken KURODA

  Dec. 2006, 食肉に関する助成研究調査成果報告書, 24, pp. 52-56, Japanese

  Others


• Simplified assay for the reproductive performance of male Japanese Black cattle by quantitative analysis of sperm phosphorylated proteins

  Hiroshi HARAYAMA, Ken KURODA

  Dec. 2005, 食肉に関する助成研究調査成果報告書, 23, pp. 39-44, Japanese

  [Refereed]

  Others


• Signal transduction system regulating head-to-head agglutination in capacitating boar spermatozoa

  Hiroshi hARAYAMA, Masashi MIYAKE, Seishiro KATO

  Dec. 2000, 食肉に関する助成研究調査成果報告書, 18, pp. 38-42, Japanese

  Others


• Molecules regulating maturational changes in mammalian spermatozoa

  Hiroshi HARAYAMA

  Lead, 2000, J. Reprod. Dev., 46(Suppl.), pp. j43-j50, Japanese

  [Refereed]

  Introduction scientific journal


• Changes of boar sperm-bound anti-agglutinin during capacitation process

  Hiroshi HARAYAMA, Masashi MIYAKE, Seishiro KATO

  Dec. 1999, 食肉に関する助成研究調査成果報告書, 17, pp. 29-33, Japanese

  Others


• Purification of seminal plasma proteins enhancing cold tolerance of boar spermatozoa and their use for sperm frozen storage

  Hiroshi HARAYAMA, Akira IMANO, Masashi MIYAKE, Seishiro KATO

  Dec. 1998, 食肉に関する助成研究調査成果報告書, 16, pp. 79-83, Japanese

  Others

• Advanced Spermatology

  Hiroshi Harayama

  Joint work, 第12 章 哺乳類精子の成熟と活性化 pp. 69～72, 筑波大学出版会, Dec. 2025, ISBN: 9784904074909


• 動物生殖科学

  原 健士朗, 原山 洋

  Joint work, 第8章 雄の配偶子形成 pp. 67～78, 朝倉書店, Feb. 2025, ISBN: 9784254450354


• 繁殖生物学 改訂版（第2版）

  原山 洋;内藤邦彦

  Joint work, 第5章3 受精 pp. 228～237, 株式会社インターズ―, Mar. 2020


• 発生生物学 「哺乳類の受精」

  原山 洋

  Joint work, 第5章 受精ー個体発生のはじまりー 5.3 哺乳類の受精 pp. 44～47, 株式会社培風館, Apr. 2019, Japanese

  Scholarly book


• Capacitation-related events in boar spermatozoa, Animal Frontier Sciences(pp. 37-43)

  Hiroshi HARAYAMA, Makiko KANETO, Masashi MIYAKE, Seishiro KATO

  Joint work, Hokuto Shobo, 2003, English

  Scholarly book


• Regulation of head-to-head agglutination in boar spermatozoa by capacitation-related factors. Reproductive Biotechnology: Reproductive biotechnology update and its related physiology (pp. 203-209)

  Hiroshi HARAYAMA, Takami MURATE, Masashi MIYAKE, Seishiro KATO

  Joint work, Hokuto Shobo, 2001, English

  Scholarly book


• Early development of porcine parthenogenetic diploids in-vitro (Reproductive Biotechnology: Reproductive biotechnology update and its related physiology. pp.285-295)

  Masashi MIYAKE, Thuan Van NGUYEN, Konosuke OKADA, Mika KATAYAMA, Statoshi KURE-BAYASHI, Hiroshi HARAYAMA

  Joint work, Hokuto Shobo, 2001, English

  Scholarly book


• Purification and characterization of boar epididymal plasma protein Anti-Agglutinin. (Reproductive Biology Update: Novel tools for assessment of environmental toxicity. pp. 207-217)

  Hiroshi HARAYAMA, Takashi MIYANO, Masashi MIYAKE, Seishiro KATO

  Joint work, Nakanishi Pub. Co., Dec. 1998, English

  Scholarly book


• Electro-activation of in vitro-matured porcine oocytes and their development in vitro and in vivo (Reproductive Biology Update: Novel tools for assessment of environmental toxicity, pp. 117-127)

  Masashi MIYAKE, Satoshi KUREBAYASHI, Takashi MIYANO, Hiroshi HARAYAMA, Seishiro KATO

  Joint work, Nakanishi Pub. Co., 1998, English

  Scholarly book

• Roles of TRPC3 in the occurrence of extracellular Ca2+-dependent full-type hyperactivation in boar sperm

  Xiaoyu Zheng, Hiroshi Harayama

  20th International Congress on Animal Reproduction (ICAR2026), Jun. 2026, English

  Poster presentation


• Relationship between coefficient of inbreeding and semen quality in Japanese Black cattle (Tajima beef cattle)

  Hiroki Matsukura, Hiromi Terada, Youko Miki, Fumiyuki Iwaki, Takayuki Akiyama, Hiroshi Harayama

  20th International Congress on Animal Reproduction (ICAR2026), Jun. 2026, English

  Poster presentation


• Roles of Ca2+ released from the internal store in the occurrence control of full-type hyperactivation of boar ejaculated sperm

  Riko Nakao, Hiroshi Harayama

  20th International Congress on Animal Reproduction (ICAR2026), Jun. 2026, English

  Poster presentation


• ウシ精子におけるFull-type Hyperactivation発生能力の個体差とcalpain 2との関係

  原山 洋, 徳永 ひな子

  日本アンドロロジー学会・第45回学術大会, Jun. 2026, Japanese

  Poster presentation


• ブタ精子でのFull-type Hyperactivationの発生制御における細胞内ストア由来Ca2+の役割

  中尾 璃子, 原山 洋

  第118回 日本繁殖生物学会大会, Sep. 2025, Japanese

  Poster presentation


• ウシ凍結保存精子でのFull-type Hyperactivationの発生状態に個体差を発生させる原因候補分子の探索

  徳永 ひな子, 吉田 早紀, 原山 洋

  第118回 日本繁殖生物学会大会, Sep. 2025, Japanese

  Poster presentation


• RXFP2 expression levels and seminal INSL3 concentrations among Japanese Black beef bull ejaculates with different levels of sperm morphological normality

  Noritoshi Kawate, H. D. A. Wimalarathne, Kenta Arashi, Fumiyuki Iwaki, Mitsuhiro Sakase, Duritahala, Hiroshi Harayama

  THe Annual Conference of Society for Study of Reproduction, 2025, Jul. 2025, English

  Poster presentation


• ウシ精子でのHyperactivation の発生能力を指標とする性状検査法の開発 － TRPC5によるHyperactivation の発生制御－

  吉田 早紀, 原山 洋

  第55回精子研究会, Jun. 2025

  [Invited]

  Nominated symposium


• ウシ凍結保存精子での鞭毛超活性化運動の発生制御へのCa2+ 依存性プロテアーゼ 「カルパイン2 」の関与

  徳永 ひな子, 原山 洋

  日本アンドロロジー学会・第44回学術大会, Jun. 2025, Japanese

  Poster presentation


• ウシ凍結保存精子でのCalyculin-A誘発性Full-type Hyperactivationの発生制御へのカルパインの関与

  徳永 ひな子, 岩木 史之, 大崎 茂, 原山 洋

  第117 回日本繁殖生物学会大会, Sep. 2024, Japanese

  Poster presentation


• ブタ精子でのFull-type Hyperactivationの誘起に及ぼす非受容体型タンパク質チロシンキナーゼの阻害剤の影響

  中尾 璃子, 植芝 愛巳, 原山 洋

  第117 回日本繁殖生物学会大会, Sep. 2024, Japanese

  Poster presentation


• ウシ精子でのCalyculin-A誘発性Full-type Hyperactivationの発生制御に機能するTRPCアイソフォームの探索

  吉田 早紀, 岩木 史之, 坂瀬 充洋, 原山 洋

  第117 回日本繁殖生物学会大会, Sep. 2024, Japanese

  Poster presentation


• ウシ凍結保存精子でのCalyculin-A 誘発性Full-type ハイパーアクチベーションの発生に及ぼすストア作動性チャネル非選択的阻害剤の影響

  吉田 早紀, 坂瀬 充洋, 原山 洋

  第73 回関西畜産学会大会, Nov. 2023, Japanese

  Oral presentation


• ウシ凍結保存精子でのCalyculin-A誘発性Full-type Hyperactivationの発生におけるストア作動性チャネルの関与

  吉田 早紀, 坂瀬 充洋, 原山 洋

  第116回日本繁殖生物学会大会, Sep. 2023, Japanese

  Oral presentation


• お父さん牛の精子がお母さんの卵子と どのように受精するのか？

  原山 洋

  第116回日本繁殖生物学会大会・市民公開講座「黒毛和牛をもっと増やそう！最新の繁殖生物学研究がお父さん牛にできること」, Sep. 2023, Japanese

  [Invited]

  Public discourse


• 人工授精での受胎率の向上を目指して －ウシ（黒毛和種）精子の新規検査法の開発－

  原山 洋

  日本アンドロロジー学会第42回学術大会 特別講演4, Jun. 2023, Japanese

  [Invited]

  Invited oral presentation


• ブタ射出精子での体外受精能力発現に及ぼす cAMP シグナリング活性化処理の影響

  原山 洋, 植芝 愛巳, 森川 莉帆, 京極 博久

  日本アンドロロジー学会第42回学術大会, Jun. 2023, Japanese

  Poster presentation


• Relationship of the SERCA activity with the time courses of occurrence of full-type hyperactivation in bovine sperm

  Duritahala, Mitsuhiro Sakase, Hiroshi Harayama

  第115回日本繁殖生物学会大会, Sep. 2022, English

  Poster presentation


• Penetration into oocytes in the medium without the PDE inhibitor by boar sperm treated with the cAMP analog and Ca2+

  植芝 愛巳, 森川 莉帆, 京極 博久, 原山 洋

  第115回日本繁殖生物学会大会, Sep. 2022, Japanese

  Poster presentation


• Involvement of cytoplasmic Ca2+ and SERCAs in the occurrence of helical movement in bovine sperm

  Duritahala, Hiroshi Harayama

  日本アンドロロジー学会第41回学術大会, Jun. 2022, English

  Poster presentation


• ウシ凍結精子でのFull-typeハイパーアクチベーションの誘起のための処理条件の検討

  原山 洋, 宮本 菜津子

  日本アンドロロジー学会第41回学術大会, Jun. 2022, Japanese

  Poster presentation


• ウシ精子において化学物質処理により誘起されるHyperactivationの特性解析

  大谷 暁洋, 坂瀬 充洋, 原山 洋

  第10回関西生殖医学集談会, Mar. 2022, Japanese

  Oral presentation


• A regulatory factor for the occurrence of helical movement in bovine cryopreserved sperm

  Duritahala, Mitsuhiro Sakase, Hiroshi Harayama

  第71回関西畜産学会, Oct. 2021, English

  Oral presentation


• Involvement of thapsigargin-sensitive Ca2+-ATPase in the occurrence of helical movement with 3-D rotation in cryopreserved bovine sperm

  Duritahala, Sakase M, Harayama H

  第114 回日本繁殖生物学会大会, Sep. 2021, English

  Poster presentation


• Thimerosal処理によりウシ精子で誘起される細胞内ストアのCa2+依存的なハイパーアクチベーションの運動様式

  大谷暁洋, 坂瀬充洋, 原山 洋

  第114 回日本繁殖生物学会大会, Sep. 2021, Japanese

  Poster presentation


• ウシ凍結精子でのFull-typeハイパーアクチベーションに及ぼすプロテインホスファターゼ阻害剤Calyculin Aの影響

  宮本菜津子, 坂瀬充洋, 原山 洋

  第114 回日本繁殖生物学会大会, Sep. 2021, Japanese

  Poster presentation


• ブタ液状保存精子でのFull-typeハイパーアクチベーションの誘起に及ぼすポリビニルアルコールおよびウシ血清アルブミンの影響

  原山 洋, 崔 聖健

  日本アンドロロジー学会第40回学術大会, Jun. 2021, Japanese

  Oral presentation


• ウシ射出精子の鞭毛運動に及ぼすNa+/K+-ATPase阻害剤Digoxinの影響

  原山 洋, Saha Soma Rani

  日本アンドロロジー学会第39回学術大会, Jan. 2021, Japanese

  Poster presentation


• ウシ精子における精密性状検査法の開発

  原山 洋

  令和2年度 東海畜産学会 シンポジウム, Dec. 2020, Japanese

  [Invited]

  Keynote oral presentation


• ブタ精子でのFull-type Hyperactivationの発生制御におけるカルモジュリンの役割

  和田篤士, 原山 洋

  第113回日本繁殖生物学会大会, Sep. 2020, Japanese

  Poster presentation


• 家畜精子の受精制御分子の探索と利用

  原山 洋

  第46回山梨大学発生工学研究センターセミナー, Dec. 2019, Japanese, Domestic conference

  [Invited]

  Public discourse


• Involvement of digoxin-sensitive Na+/K+-ATPase in the regulation of bovine sperm motility

  SAHA Soma Rani, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  第112回日本繁殖生物学会, Sep. 2019, English, Domestic conference

  Poster presentation


• Characterization of SPACA1 proteins in the spermatozoa from Japanese Black bulls

  MINAMI Kenta, ASO Miyuki M, YAMADA Ayano, KISHIDA Kazumi, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  第112回日本繁殖生物学会, Sep. 2019, Japanese, Domestic conference

  Poster presentation


• 低繁殖症の雄ウシを検出するための精子分子性状検査法のマーカー「SPACA1タンパク質」の解析

  MINAMI Kenta, ASO Miyuki, YAMADA Ayano, KISHIDA Kazumi, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  日本アンドロロジー学会第38回学術大会, Jun. 2019, Japanese, 大阪国際会議場（グランキューブ大阪）, Domestic conference

  Poster presentation


• Investigation of movement patterns of bull ejaculated sperm to improve the evaluation method of motility

  YAMADA Ayano, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  第111回日本繁殖生物学会大会, Sep. 2018, Japanese, 上田市 信州大学繊維学部, Domestic conference

  Poster presentation


• cAMP-PKAシグナル伝達活性の違いがウシ精子の運動様式に及ぼす影響 － 精子の活力検査における評価基準の見直し －

  YAMADA Ayano, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  日本アンドロロジー学会第37回学術大会, Jun. 2018, Japanese, ラ・スイート神戸オーシャンガーデン, Domestic conference

  Poster presentation


• ウシ精子での細胞外Ca2+依存的なFull-typeハイパーアクチベーションの抑制に機能するカリクリンA感受性プロテインホスファターゼアイソフォームの特定

  ARAI Yuka, HARAYAMA Hiroshi

  第6回関西生殖医学集談会, Feb. 2018, Japanese, 大阪市 ハービスPLAZA, Domestic conference

  Oral presentation


• Mechanism for the extracellular Ca2+-dependent occurrence of full-type hyperactivation in boar sperm treated with a cAMP analog

  OTSUKA Nagisa, HARAYAMA Hiroshi

  Fourth World Congress of Reproductive Biology (WCRB2017), Sep. 2017, English, 沖縄県宜野湾市 沖縄コンベンションセンター, International conference

  Poster presentation


• Calyculin A-sensitive protein phosphatases which are involved in the suppression for full-type hyperactivation of bovine sperm

  ARAI Yuka, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  Fourth World Congress of Reproductive Biology (WCRB2017), Sep. 2017, English, 沖縄県宜野湾市 沖縄コンベンションセンター, International conference

  Poster presentation


• ブタ精子での超活性化運動（Full-type HA）の開始におけるTRPC3チャネルの役割

  OTSUKA Nagisa, HARAYAMA HIROSHI

  第5回関西生殖医学集談会, Mar. 2017, Japanese, 大阪市, Domestic conference

  Oral presentation


• Advanced techniques for the reproduction in the cattle

  HARAYAMA Hiroshi

  UGSVS lecture, Feb. 2017, English, 岐阜大学, Domestic conference

  [Invited]

  Invited oral presentation


• ブタ精子におけるfull-type hyperactivationの開始制御へのTRPC3チャネルの関与

  OTSUKA Nagisa, HARAYAMA Hiroshi

  第109回日本繁殖生物学会, Sep. 2016, Japanese, 一社 日本繁殖生物学会, 相模原市（麻布大学）, Domestic conference

  Poster presentation


• ウシ精子のハイパーアクチベーションを抑制するカリクリンA感受性プロテインホスファターゼの特定

  OTSUKA Nagisa, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  第109回日本繁殖生物学会, Sep. 2016, Japanese, 一社 日本繁殖生物学会, 相模原市（麻布大学）, Domestic conference

  Poster presentation


• 先体タンパク質をマーカーとする哺乳類精子の分子性状検査法

  HARAYAMA HIROSHI

  日本アンドロロジー学会35回学術大会 シンポジウム1：男性不妊症の分子メカニズム, Jun. 2016, Japanese, 前橋市・前橋テルサ, Domestic conference

  [Invited]

  Nominated symposium


• ブタ精子での先体反応誘起に伴う頭部タンパク質SPACA1の変化

  OGURA YUKARI, HARAYAMA HIROSHI

  日本畜産学会第121回大会, Mar. 2016, Japanese, 日本獣医生命大学, Domestic conference

  Poster presentation


• ウシ新鮮射出精子における先体チロシンリン酸化タンパク質の分布状態

  ARAI M. MIYUKI, HARAYAMA HIROSHI

  第4回関西生殖医学集談会・第48回関西アンドロロジーカンファレンス合同研究会, Mar. 2016, Japanese, 大阪市, Domestic conference

  Oral presentation


• ウシ新鮮射出精子の分子性状における個体差 －先体チロシンリン酸化タンパク質と人工授精成績との関係－

  ARAI Miyuki, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  第108回日本繁殖生物学会大会, Sep. 2015, Japanese, 日本繁殖生物学会, 宮崎, Domestic conference

  Poster presentation


• 哺乳類の精子学 ー運動の開始から受精までー

  HARAYAMA HIROSHI

  第35回関西生殖発生毒性フォーラム 基礎教育講演, Apr. 2015, Japanese, 大阪市, Domestic conference

  [Invited]

  Invited oral presentation


• Mammalian sperm acrosomal protein affecting the artificial insemination results and the in-vitro fertilization results

  HARAYAMA HIROSHI

  第60回日本生殖医学会学術講演会 シンポジウム3 「受精に関する最近の話題」, Apr. 2015, Japanese, 横浜, Domestic conference

  [Invited]

  Nominated symposium


• ウシ凍結精子での先体損傷・離脱に伴うIZUMO1の変化

  FUKUDA Masaki, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  日本畜産学会第119回大会, Mar. 2015, Japanese, 宇都宮, Domestic conference

  Oral presentation


• 低繁殖症雄ウシの精子における欠陥型タンパク質の検出および特性解析

  HARAYAMA HIROSHI

  受胎率向上SIG第1回会合, Dec. 2014, Japanese, 東京, Domestic conference

  [Invited]

  Nominated symposium


• The association between spatial distribution patterns of SPACA1 in human ejaculated sperms and outcomes of conventional IVF

  KISHIDA Kazumi, HARAYAMA Hiroshi, KIMURA Fuminori, MURAKAMI Takashi

  World Congress of Reproductive Biology 2014, Sep. 2014, English, Edinburgh, UK, International conference

  Poster presentation


• 黒毛和種精子でのチロシンリン酸化タンパク質の分布状態が先体の安定性に及ぼす影響

  ARAI Miyuki, MINAMI Kenta, OGURA Yukari, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  第107回日本繁殖生物学会大会, Aug. 2014, Japanese, 日本繁殖生物学会, 帯広, Domestic conference

  Poster presentation


• ブタ胚盤胞におけるタイトジャンクション関連タンパク質claudin familyの発現性

  MIYAKE Masashi, TOU Tetsu, SHIBUTANI Mihiro, LEE Jibak, HARAYAMA Hiroshi, MIYANO Takashi

  第107回日本繁殖生物学会大会, Aug. 2014, Japanese, 日本繁殖生物学会, 帯広, Domestic conference

  Oral presentation


• ブタ精子での先体反応に伴う頭部タンパク質「SPACA1」の分布および分子マスの変化

  OGURA Yukari, TAKAGISHI Yuki, KOJIMA Aya, ISHIKAWA Sho, HARAYAMA Hiroshi

  第107回日本繁殖生物学会大会, Aug. 2014, Japanese, 日本繁殖生物学会, 帯広, Domestic conference

  Poster presentation


• ウシ凍結保存精子における先体の損傷・離脱が頭部でのIZUMO1の分布に及ぼす影響

  FUKUDA Masaki, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  第107回日本繁殖生物学会大会, Aug. 2014, Japanese, 日本繁殖生物学会, 帯広, Domestic conference

  Poster presentation


• ウシ精巣におけるcAMP依存性転写調節因子CREMの発現パターンの解析

  NODA Taichi, MINAMI Kenta, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  第107回日本繁殖生物学会大会, Aug. 2014, Japanese, 日本繁殖生物学会, 帯広, Domestic conference

  Poster presentation


• 家畜精子のハイパーアクチベーションに先立つ部位特異的なキャパシテーション

  HARAYAMA HIROSHI

  日本アンドロロジー学会第33回学術大会 シンポジウム 1 「Capacitation の最前線を極める」, Jun. 2014, Japanese, 軽井沢プリンスホテルウエスト, Domestic conference

  [Invited]

  Nominated symposium


• ブタ精子の鞭毛に分布するカルパイン2の役割

  KOJIMA Aya, HARAYAMA Hiroshi

  第2回関西生殖医学集談会, Mar. 2014, Japanese, 兵庫医科大学産婦人科学講座, 大阪市, Domestic conference

  Oral presentation


• ヒト射出精子の頭部におけるSPACA1の分布状態の個体差 －先体主部での分布状態と体外受精成績との関係－

  KISHIDA Kazumi, HARAYAMA Hiroshi, KIMURA Fuminori, MURAKAMI Takashi

  第2回関西生殖医学集談会, Mar. 2014, Japanese, 兵庫医科大学産婦人科学講座, 大阪市, Domestic conference

  Oral presentation


• Full-type hyperactivationの誘起に伴う家畜精子の頸部および中片部における変化

  HARAYAMA Hiroshi, MIZUNO Yohei, KOJIMA Aya

  日本畜産学会第118回大会, Mar. 2014, Japanese, 日本畜産学会, つくば, Domestic conference

  Oral presentation


• 人工授精で雌を受胎させにくい精子を産生する黒毛和種雄個体の検出法 －凍結保存後の精子先体の正常性と体内受精由来の移植可能胚率の関係－

  SAKASE Mitsuhiro, KOJI Reiko, KOHAMA Minako, AKIYAMA Yoshitaka, OKA Akio, HARAYAMA Hiroshi, FDUKUSHIMA Moriyuki

  第106回日本繁殖生物学会大会, Sep. 2013, Japanese, 日本繁殖生物学会, 府中市, Domestic conference

  Oral presentation


• ブタ精子におけるカルパインの検出および機能解析

  KOJIMA AYA, ISHIKAWA Sho, SHIDARA Osamu, HARAYAMA Hiroshi

  第106回日本繁殖生物学会大会, Sep. 2013, Japanese, 日本繁殖生物学会, 府中市, Domestic conference

  Poster presentation


• ヒト射出精子におけるSPACA1の検出 －先体での検出パターンと体外受精成績との相関性－

  KISHIDA Kazumi, HARAYAMA Hiroshi, KIMURA Fuminori, MURAKAMI Takashi

  第106回日本繁殖生物学会大会, Sep. 2013, Japanese, 日本繁殖生物学会, 府中市, Domestic conference

  Poster presentation


• ウシ精子の鞭毛におけるタンパク質チロシンリン酸化および超活性化運動の誘起

  MIZUNO Yohei, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  関西畜産学会第63回大会, Sep. 2013, Japanese, 関西畜産学会, 彦根市, Domestic conference

  Oral presentation


• ブタ精子での後帽部タンパク質の脱リン酸化および先体反応におけるcAMP-EPACシグナリングの役割

  ISONO Ayane, KOJIMA Aya, TAKAGISHI Yuki, ISHIKAWA Sho, SHIDARA Osamu, HARAYAMA Hiroshi

  日本畜産学会第116回大会, Mar. 2013, Japanese, 日本畜産学会, 広島市, Domestic conference

  Oral presentation


• ウシのハイパーアクチベーション精子における鞭毛タンパク質のリン酸化状態

  MIZUNO Yohei, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  日本畜産学会第116回大会, Mar. 2013, Japanese, 日本畜産学会, 広島市, Domestic conference

  Oral presentation


• 哺乳類精子の細胞内cAMPシグナル伝達機構に関する研究

  HARAYAMA HIROSHI

  第105回日本繁殖生物学会大会, Sep. 2012, Japanese, 日本繁殖生物学会, つくば市, Domestic conference

  [Invited]

  Invited oral presentation


• 黒毛和種精子先体前部におけるチロシンリン酸化型SPACA1の解析－サンプル間差および精子成熟に伴う変化について－

  MINAMI Kenta, NODA Taichi, ISONO Ayane, KOJIMA Aya, MIZUNO Yohei, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  第105回日本繁殖生物学会大会, Sep. 2012, Japanese, 日本繁殖生物学会, つくば市, Domestic conference

  Poster presentation


• 黒毛和種精子における先体の耐凍性マーカーの特性解析

  MINAMI Kenta, NODA Taichi, ISONO Ayane, KOJIMA Aya, MIZUNO Yohei, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA HIROSHI

  第62回関西畜産学会大会, Sep. 2012, Japanese, 関西畜産学会, 和歌山市, Domestic conference

  Oral presentation


• cAMP合成酵素ADCY10の異常型スプライスバリアントの検出によるウシ精子の新規の性状検査法

  NODA Taichi, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA HIROSHI

  第105回日本繁殖生物学会大会, Sep. 2012, Japanese, 日本繁殖生物学会, つくば市, Domestic conference

  Oral presentation


• ブタの精巣および射出精子における活性型CREM（cAMP-responsive element modulator）の分子性状の解析

  NODA Taichi, SHIDARA Osamu, ISHIKAWA Sho, HARAYAMA Hiroshi

  日本アンドロロジー学会第31回学術大会, Jun. 2012, Japanese, 日本アンドロロジー学会, 神戸市, Domestic conference

  Oral presentation


• 黒毛和種精子における耐凍能マーカーの検出パターン

  MINAMI Kenta, NODA Taichi, ISONO Ayane, KOJIMA Aya, MIZUNO Yohei, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA HIROSHI

  日本畜産学会第115回大会, Mar. 2012, Japanese, 日本畜産学会, 名古屋市, Domestic conference

  Oral presentation


• ブタ雄性生殖細胞での活性型CREMの検出および分布解析

  NODA Taichi, SHIDARA Osamu, HARAYAMA HIROSHI

  日本畜産学会第115回大会, Mar. 2012, Japanese, 日本畜産学会, 名古屋市, Domestic conference

  Oral presentation


• ウシ精子における断片型ADCY10の合成メカニズム

  NODA Taichi, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA HIROSHI

  第137回日本生殖医学会関西支部集談会, Mar. 2012, Japanese, 大阪市, Domestic conference

  Oral presentation


• ウシの可溶化型アデニル酸シクラーゼ (ADCY10) の断片型は精巣においてmRNAスプライスバリアントから合成される

  NODA Taichi, WATAI Yusuke, SAKASE Mitsuhiro, KOHAMA Namiko, FUKUSHIMA Moriyuki, HARAYAMA HIROSHI

  第104回日本繁殖生物学会大会, Sep. 2011, Japanese, 日本繁殖生物学会, 盛岡市, Domestic conference

  Poster presentation


• ブタ精子でのcAMP依存的な鞭毛超活性化運動に及ぼすカルモジュリンアンタゴニストおよびカルパイン阻害剤

  HARAYAMA Hiroshi, NODA Taichi, SHIDARA Osamu

  第136回日本生殖医学会関西支部集談会, Mar. 2011, Japanese, 大阪市, Domestic conference

  Oral presentation


• 哺乳類精子における部位特異的なcAMPシグナリング

  HARAYAMA HIROSHI

  日本動物学会第80回大会「第8回受精シンポジウム」, Sep. 2010, Japanese, 日本動物学会, 静岡市, Domestic conference

  [Invited]

  Nominated symposium


• ブタ精子中片部のAMPKはcAMP依存性ハイパーアクチベーションの発現制御に関与する

  HARAYAMA Hiroshi, NODA Taichi, SHIDARA Osamu

  日本アンドロロジー学会第29回学術大会, Jul. 2010, Japanese, 日本アンドロロジー学会, 東京, Domestic conference

  Oral presentation


• Paternal exposure to 2,3,7,8-tetrachloro- dibenzo-p-dioxin (TCDD) affects the sex ratio of offspring at birth by change the sex ratio of fertilized egg

  ISHIHARA KANA, TASAKA KEN, TANIDA TAKASHI, WARITA KATSUHIKO, MITSUHASHI TOMOKO, MIYAKE MASASHI, HARAYAMA HIROSHI, YOKOYAMA TOSHIFUMI, KITAGAWA HIROSHI, HOSHI NOBUHIKO

  11th International Symposium on Spermatology, Jun. 2010, English, 宜野湾市, International conference

  Poster presentation


• Paternal exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) affects the sex ratio of offspring at birth by change the sex ratio of fertilized egg

  ISHIHARA K, TASAKA K, TANIDA T, WARITA K, MITSUHASHI T, MIYAKE M, HARAYAMA Hiroshi, YOKOYAMA T, KITAGAWA H, HOSHI N

  11th International Symposium on Spermatology, Jun. 2010, English, Naha, International conference

  Poster presentation


• Immunodetection patterns of tyrosine-phosphorylated proteins in the head of frozen-thawed spermatozoa of Japanese Black bulls

  HARAYAMA Hiroshi, MURASE Tetsuma, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki

  11th International Symposium on Spermatology, Jun. 2010, English, Naha, International conference

  Public symposium


• Identification of cAMP responsive element modulator (CREM) isoforms of the porcine testis

  NODA Taichi, SHIDARA Osamu, HARAYAMA Hiroshi

  11th International Symposium on Spermatology, Jun. 2010, English, Naha, International conference

  Poster presentation


• ブタ精巣におけるcAMP依存性転写調節因子CREMτの発現

  NODA Taichi, SHIDARA Osamu, HARAYAMA Hiroshi

  日本畜産学会第112回大会, Mar. 2010, Japanese, 日本畜産学会, 東京, Domestic conference

  Oral presentation


• ブタの精子形成関連タンパク質の発現を制御するcAMPシグナリング構成分子の同定

  NODA Taichi, HARAYAMA Hiroshi

  第135回日本生殖医学会関西支部集談会, Mar. 2010, Japanese, 大阪市, Domestic conference

  Oral presentation


• ウシ精巣におけるアデニル酸シクラーゼ10(ADCY10)mRNAsのスプライスバリアントの検出

  WATAI Yusuke, NODA Taichi, SAKASE Mitsuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  第135回日本生殖医学会関西支部集談会, Mar. 2010, Japanese, 大阪市, Domestic conference

  Oral presentation


• Subfertility in Japanese Black bulls and new methods to detect abnormal functionality of spermatozoa

  MURASE Tetsuma, MUKOUJIMA K, HARAYAMA HIROSHI, HOSHINO Y, KATO T

  International Symposium of Declining Fertility in Dairy Cows in the World; its Causes and Possible Solutions, Feb. 2010, English, Miyazaki, International conference

  Poster presentation


• Active protein kinase A (PKA) is required for the meiotic resumption of pig oocytes

  CAYO-COLCA I S, HARAYAMA Hiroshi, MIYANO Takashi

  2009 Japan-Taiwan Joint Symposium on Cell Signaling and Gene Regulation, Nov. 2009, English, Kobe, International conference

  Poster presentation


• ブタ単為発生2倍体の胚盤胞への発生に及ぼすヘキソースの影響

  YONEZAWA Jun-ichi, HARAYAMA Hiroshi, MIYAKE Masashi

  第102回日本繁殖生物学会大会, Sep. 2009, Japanese, 日本繁殖生物学会, 奈良市, Domestic conference

  Poster presentation


• Changes in protein kinase A (PKA) activity during meiotic resumption of pig oocytes

  Cayo-Colca I S, HARAYAMA Hiroshi, MIYANO Takashi

  第102回日本繁殖生物学会大会, Sep. 2009, English, 日本繁殖生物学会, 奈良市, Domestic conference

  Poster presentation


• ブタ精巣におけるcAMP依存性転写調節因子(CREM)ファミリーアイソフォームの発現

  NODA Taichi, SHIDARA Osamu, HARAYAMA Hiroshi

  日本アンドロロジー学会第28回学術大会, Jul. 2009, Japanese, 日本アンドロロジー学会, 富山市, Domestic conference

  Oral presentation


• ブタ精子におけるADCY10の検出

  NAKAMURA Kazumi, SUZUKI Chihiro, TATE Shunsuke, LEE J, HARAYAMA HIROSHI

  日本アンドロロジー学会第28回学術大会, Jul. 2009, Japanese, 日本アンドロロジー学会, 富山市, Domestic conference

  Oral presentation


• Detection of soluble adenylyl cyclase (ADCY10) homolog proteins in boar spermatozoa

  NAKAMURA Kazumi, SUZUKI Chihiro, TATE Shunsuke, LEE Jibak, HARAYAMA Hiroshi

  34th Annual Meeting of the American Society of Andrology, Apr. 2009, English, The American Society of Andrology, Philadelphia, International conference

  Poster presentation


• ブタ精子でのハイパーアクチベーションの発現制御における解糖系の役割

  HARAYAMA Hiroshi, NAKAMURA Kazumi, MIYAKE Masashi

  日本畜産学会第110回大会, Mar. 2009, Japanese, 日本畜産学会, 藤沢市, Domestic conference

  Oral presentation


• cAMPアナログ処理により鞭毛超活性化運動が誘起されたブタ精子でのAMPKサブユニットの分子変化

  HARAYAMA Hiroshi, NODA Taichi, MIYAKE Masashi

  第134回日本生殖医学関西支部集談会, Mar. 2009, Japanese, 大阪市, Domestic conference

  Oral presentation


• 哺乳類精子の頚部特異的なcAMPシグナリングによるハイパーアクチベーションの制御

  HARAYAMA Hiroshi

  第40回精子研究会シンポシウム, Jan. 2009, Japanese, 精子研究会, 吹田市, Domestic conference

  [Invited]

  Nominated symposium


• 低受胎率を示した黒毛和種あるいは正常な個体における凍結－融解精子鞭毛の超活性化運動

  MURASE Tetsuma, Ismail EI-kon, MUKAIJIMA Koji, HARAYAMA Hiroshi, TAKASU Masaki, SAKAI Kenji

  第101回日本繁殖生物学会大会, Sep. 2008, Japanese, 日本繁殖生物学会, 福岡市, Domestic conference

  Oral presentation


• cAMP-Epacシグナリングを介したブタ精子キャパシテーションの新規制御メカニズム

  NAKAMURA Kazumi, TATE Shunsuke, HARAYAMA HIROSHI

  日本アンドロロジー学会第27回学術大会, Jul. 2008, Japanese, 日本アンドロロジー学会, 京都市, Domestic conference

  Oral presentation


• Cyclic AMP-induced inactivation of PDK1 is linked to the enhancement of protein tyrosine phosphorylation and flagellar hyperactivation in boar spermatozoa

  HARAYAMA Hiroshi

  33rd Annual Meeting of American Society of Andrology, Apr. 2008, English, American Society of Andrology, Albuquerque, International conference

  Poster presentation


• ウシ精子のハイパーアクチベーション発現制御に関与する候補分子の特定

  NISHIJIMA Kazuhiro, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  第109回日本畜産学会大会, Mar. 2008, Japanese, 日本畜産学会, 水戸市, Domestic conference

  Oral presentation


• マウス精巣上体尾精子の鞭毛主部においてプロテインホスファターゼはPKA活性を抑制する

  Namiko GOTO, Hiroshi HARAYAMA

  The 100th Annual meeting of the Japanese Society of Animal Reproduction, Oct. 2007, Japanese, 日本繁殖生物学会, Tokyo, Domestic conference

  Oral presentation


• ブタ精子頭部でのcAMP依存性細胞内カルシウム濃度上昇およびキャパシテーションにおけるEpacの役割

  Kazumi NAKAMURA, Shunsuke TATE, Osamu SHIDARA, Hiroshi HARAYAMA

  The 100th Annual meeting of the Japanese Society of Animal Reproduction, Oct. 2007, Japanese, 日本繁殖生物学会, Tokyo, Domestic conference

  Oral presentation


• ブタ精子の部位特異的な細胞内cAMPシグナリング：ハイパーアクチベーション誘起に伴う鞭毛主部でのPKAおよびPDK1のcAMP依存的な変化

  HARAYAMA Hiroshi, TATE Shunnsuke, NAKAMURA Kazumi, SHIDARA Osamu, MIYAKE Masashi

  The 100th Annual meeting of the Japanese Society of Animal Reproduction, Oct. 2007, Japanese, 日本繁殖生物学会, Tokyo, Domestic conference

  Poster presentation


• ウシ，ブタおよびマウスの精子でのタンパク質チロシンリン酸化に及ぼすcAMPアナログとタンパク質チロシンホスファターゼの影響

  NISHIJIMA Kazuhiro, GOTO Namiko, NAKAMURA Kazumi, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  The 100th Annual meeting of the Japanese Society of Animal Reproduction, Oct. 2007, Japanese, Tokyo, Domestic conference

  Poster presentation


• ブタ精子の部位特異的な細胞内cAMPシグナリング：頚部のcAMP-PKA-PKCシグナリングはハイパーアクチベーションの発現調節に関与する

  Hiroshi HARAYAMA, Masashi MIYAKE, Osamu SHIDARA

  日本アンドロロジー学会第26回学術大会, Jul. 2007, Japanese, 浦安市, Domestic conference

  Oral presentation


• ウシおよびブタの精子におけるcAMP依存性タンパク質チロシンリン酸化

  Kazuhiro NISHIJIMA, Namiko GOTO, Moriyuki FUKUSHIMA, Hiroshi HARAYAMA

  日本アンドロロジー学会第26回学術大会, Jul. 2007, Japanese, 浦安市, Domestic conference

  Oral presentation


• 人間との共生環境の回復を目的とした有害野生動物の繁殖抑制

  HARAYAMA Hiroshi, GOTO Namiko, TATE Shunsuke

  環境科学総合研究所成果報告会, May 2007, Japanese, 熱海, Domestic conference

  Public discourse


• 凍結融解処理されたブタ精子における32 kDaチロシンリン酸化タンパク質Tyr32の出現メカニズム

  Hiroshi HARAYAMA, Tomohito TABUCHI, Osamu SHIDARA

  日本畜産学会第107回大会, Mar. 2007, Japanese, 相模原市, Domestic conference

  Oral presentation


• ブタ着床前胚におけるタイトジャンクション関連タンパク質―Occludin の発現性

  許 尚丹, HARAYAMA Hiroshi, MIYAKE Masashi

  第100回日本繁殖生物学会大会, Mar. 2007, Japanese, 日本繁殖生物学会, 東京, Domestic conference

  Poster presentation


• ブタ精子のキャパシテーションにおけるcAMP-Epac-カルシウムシグナリングの役割

  Shun-suke TATE, Kazumi NAKAMURA, Hiroshi HARAYAMA

  日本畜産学会第107回大会, Mar. 2007, Japanese, 相模原市, Domestic conference

  Oral presentation


• Localization of claudin family proteins in pig embryos during preimplantation development

  S. XU, J. LEE, Hiroshi HARAYAMA, Masashi MIYAKE

  33rd Annual Conference of the International Embryo Transfer Society, Jan. 2007, English, Kyoto, JAPAN, International conference

  Poster presentation


• Fertilization-related parameters of frozen-thawed spermatozoa from subfertile Japanese Black cattle

  Ken KURODA, Moriyuki FUKUSHIMA, Masashi MIYAKE, Hiroshi HARAYAMA

  33rd Annual Conference of the International Embryo Transfer Society, Jan. 2007, English, Kyoto, Japan, International conference

  Poster presentation


• Assembly of nuclear membrane proteins in bovine IVF and activated oocytes.

  MIYAKE Masashi, ISAJI Mamiko, HARAYAMA Hiroshi

  The 3rd Asian Reproductive Biotechnology Conference, Nov. 2006, English, Asian Reproductive Biotechnology Conference, Hanoi, Vietnam, International conference

  Invited oral presentation


• Detection of cAMP-Epac signaling cascades in mouse epididymal spermatozoa

  GOTO Namiko, AMANO Reiko, LEE Jibak, HARAYAMA Hiroshi

  99th Annual meeting of Japanese Society of Animal Reproduction, Sep. 2006, Japanese, 日本繁殖生物学会, Nagoya, Domestic conference

  Poster presentation


• cAMP-PKA signaling-dependent activation of PKC is involved in regulation of flagellar hyperactivation in boar spermatozoa

  HARAYAMA Hiroshi, MIYAKE Masashi, SHIDARA Osamu

  99th Annual Meeting of Japanese Society of Animal Reproduction, Sep. 2006, Japanese, Japanese Society of Animal Reproduction, Nagoya, Domestic conference

  Oral presentation


• Localization of proteins related to the tight junction in the pig and mouse preimplantational embryos.

  XU Shangdan, KIMURA Hayato, MATSUMURA Hirokazu, LEE Jibak, HARAYAMA Hiroshi, MIYAKE Masashi

  The 99th Annual meeting of the Japanese Society of Animal Reproduction, Sep. 2006, Japanese, 日本繁殖生物学会, Nagoya, Domestic conference

  Poster presentation


• Nuclear localization of a tight Junction protein claudin-1 in the preimplantation embryos

  XU Shangdan, KIMURA Hayato, Matsumura Hirokazu, LEE Jibak, HARAYAMA Hiroshi, MIYAKE Masashi

  The 13th Annual Meeting of Japan Embryo Transfer Society, Aug. 2006, Japanese, 日本胚移植研究会, Hiroshima, Domestic conference

  Oral presentation


• Effects of sodium bicarbonate and PKA inhibitor on cAMP-dependent elevation of intracellular calcium concentration in the boar sperm head

  TATE Shunsuke, SHIDARA Osamu, HARAYAMA Hiroshi

  56th Annual meeting of Kansai Society of Animal Science, Aug. 2006, Japanese, 関西畜産学会, Osaka, Domestic conference

  Oral presentation


• Role of protein kinase C in the cyclic adenosine 3',5'-monophosphate-dependent hyperactivation of boar spermatozoa

  HARAYAMA Hiroshi, MIYAKE Masashi

  31st Annual meeting of American Society of Andrology, Apr. 2006, English, American Society of Andrology, Chicago, USA, International conference

  Poster presentation


• Premature capacitation and acrosome reaction in frozen spermatozoa from subfertile Japanese Black cattle

  KURODA Ken, FUKUSHIMA Moriyuki, HARAYAMA Hiroshi

  106th Annual meeting of Japanese Society of Animal Science, Mar. 2006, Japanese, 日本畜産学会, Fukuoka, Domestic conference

  Poster presentation


• Effects of calyculin A on cAMP-dependent decrease of Ser-/Thr-phosphorylated proteins in the postacrosomal region of boar spermatozoa

  ADACHI Jun, HARAYAMA Hiroshi, MIYAKE Masashi

  106th Annual meeting of Japanese Society of Animal Science, Mar. 2006, Japanese, 日本畜産学会, Fukuoka, Domestic conference

  Poster presentation


• Detection of Ca2+ store marker proteins in boar and mouse spermatozoa

  ONO Mikiko, HARAYAMA Hiroshi, MIYAKE Masashi

  106th Annual meeting of Japanese Society of Animal Science, Mar. 2006, Japanese, 日本畜産学会, Fukuoka, Domestic conference

  Oral presentation


• Low progressive motility and premature expression of fertilizing ability of frozen-thawed spermatozoa from subfertile Japanese Black cattle

  KURODA Ken, HARAYAMA Hiroshi

  Kobe University The 21st Century COE Program Symposium: Japan-Taiwan Symposium on Cell Signaling and Gene Expression (Program and Abstract,18), Nov. 2005, English, Kobe University COE Program "Signaling Mechanisms by Protein Modification Reactions", Kobe, International conference

  Oral presentation


• Low progressive motility and premature capacitation-acrosome reaction in the frozen-thawed spermatozoa from subfertile Japanese Black cattle

  KURODA Ken, FUKUSHIMA Moriyuki, 原山 洋

  98th Annual Meeting of Japanese Society of Animal Reproduction, Sep. 2005, Japanese, 日本繁殖生物学会, Shizuoka, Domestic conference

  Poster presentation


• Effects of PKA inhibitor on the cAMP-dependent elevation of intracellular calcium concentration in boar sperm head

  TATE Shunsuke, 設楽 修, 原山 洋

  98th Annual Meeting of Japanese Society of Animal Reproduction, Sep. 2005, Japanese, 日本繁殖生物学会, Shizuoka, Domestic conference

  Oral presentation


• cAMP-dependent changes in glycochains and phosphoproteins in the postacrosomal region of boar spermatozoa

  ADACHI Jun, Tate Shunsuke, 原山 洋, 三宅 正史

  2005 (55th) Annual Meeting of Kansai Society of Animal Science, Sep. 2005, Japanese, 関西畜産学会, Matsuyama, Domestic conference

  Oral presentation


• Low progressive motility, low PKA activity and premature capacitation of frozen-thawed spermatozoa from subfertile bulls

  KURODA Ken, HARAYAMA Hiroshi

  8th International Congress of Andrology (Int. J. Androl., Vol. 28(Suppl. 1), 79), Jun. 2005, English, International Society of Andrology, Seoul, Korea, International conference

  Poster presentation


• A cyclic AMP-dependent tyrosine phosphorylation of phospholipase C in boar spermatozoa

  Harayama Hiroshi

  8th International Congress of Andrology (Int. J. Androl., Vol. 28(Suppl. 1), 78), Jun. 2005, English, International Society of Andrology, Seoul, Korea, International conference

  Oral presentation


• Analysis of the cAMP signaling cascade in mouse spermatozoa

  IWASA Chika, 原山 洋

  104th Annual Meeting of Japanese Society of Animal Science, Mar. 2005, Japanese, 日本畜産学会, Tokyo, Domestic conference

  Oral presentation


• cAMP-dependent activation of phospholipase Cgamma in boar spermatozoa

  原山 洋, 三宅 正史, Murase Tetsuma

  104th Annual Meeting of Japanese Society of Animal Science, Mar. 2005, Japanese, 日本畜産学会, Tokyo, Domestic conference

  Oral presentation


• The activation of boar sperm tyrosine kinase SYK is regulated via calcium-independent cAMP signaling

  原山 洋

  82th Annual Meeting of Japanese Society of Swine Science, Oct. 2004, Japanese, 日本養豚学会, National Livestock Breeding Center, Domestic conference

  Oral presentation


• cAMP-dependent activation of non-receptor tyrosine kinase SYK in boar spermatozoa

  原山 洋, MUROGA Mai, 三宅 正史

  97th Annual Meeting of Japanese Society of Animal Reproduction, Sep. 2004, Japanese, 日本繁殖生物学会, Hiroshima, Domestic conference

  Poster presentation


• Phosphorylation of PKA substrate proteins in boar capacitating spermatozoa

  KURODA Ken, 原山 洋

  97th Annual Meeting of Japanese Society of Animal Reproduction, Sep. 2004, Japanese, 日本繁殖生物学会, Hiroshima, Domestic conference

  Oral presentation


• Formation of nuclear envelope and dephosphorylation of histone H3 in the in-vitro fertilized bovine oocytes

  ISAJI Mamiko, IWATA Hisataka, HARAYAMA Hiroshi, Lee Jibak, MIYAKE Masashi

  The 11th Annual Meeting of Japan Embryo Transfer Society, Aug. 2004, Japanese, 日本胚移植研究会, Matsuyama, Domestic conference

  Oral presentation


• cAMP-induced increase of 32-kDa tyrosine-phosphorylated protein (TyrP32) is related to the calcium-dependent disintegration of the acrosome in boar sperm.

  HARAYAMA Hiroshi, SASAKI Kiyomi, MIYAKE Masashi

  The American Society of Andrology, 29th Annual Meeting., Apr. 2004, English, The American Society of Andrology, USA, Baltimore, International conference

  Poster presentation


• ウシ受精卵母細胞におけるLys9でメチル化されているヒストンH3の局在性

  伊佐治 麻実子, 岩田 尚孝, 原山 洋, 三宅 正史

  The 103th Annual meeting of Animal Science, Mar. 2004, Japanese, 日本畜産学会, 東京農工大学, Domestic conference

  Oral presentation


• Role of calcium in the cAMP-induced increase of TyrP32 and acrosomal disintegration

  原山 洋, SASAKI Kiyomi, 三宅 正史

  103th Annual Meeting of Japanese Society of Animal Science, Mar. 2004, Japanese, 日本畜産学会, Tokyo University of Agriculture and Technology, Domestic conference

  Oral presentation


• ウシ受精ならびに活性化卵母細胞におけるLAP2βの局在とその凝集時期

  伊佐治 麻実子, 岩田 尚孝, 原山 洋, 三宅 正史

  第27回日本分子生物学会年会, 2004, Japanese, 日本分子生物学会, 未記入, Domestic conference

  Oral presentation


• Viability and protein phosphorylation state of boar sperm undergoing cAMP analogue-induced head-to-head agglutination

  HARAYAMA Hiroshi, SASAKI K, MIYAKE Masashi

  The AmericanSociety of Andrology 28th Annual Meeting,, 2003, English, 未記入, Washington. DC, International conference

  Oral presentation


• Localization of LAP2beta in oocytes and embryos during early development in cattle, The Annual Conference of the International Embryo Transfer Society

  ISAJI M, IWATA H, OHTA M, HARAYAMA Hiroshi, MIYAKE Masashi

  Theriogenology, 2003, English, 未記入, 未記入, International conference

  Oral presentation


• Behavior of chromatins in pronuclei in the cattle and mouse

  ISAJI M, IWATA H, OHTA M, HAYARAMA Hiroshi, MIYAKE Masashi

  36th Annual Meeting of the Society for the Study of Reproduction, 2003, English, 未記入, 未記入, International conference

  Oral presentation

• 関西アンドロロジーカンファレンス


• 関西生殖医学集談会


• 精子研究会


• 日本生殖医学会


• The Society for the Study of Reproduction


• 関西畜産学会


• 日本アンドロロジー学会


• 日本繁殖生物学会


• 日本畜産学会

• Elucidation of roles and signal transduction of INSL3 in bovine gamate maturaiton, fertilization and embryo development

  川手 憲俊, 原山 洋, 古山 敬祐

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (C), Osaka Metropolitan University, 01 Apr. 2024 - 31 Mar. 2027


• Establishment and spread of the new assay for sperm flagellar function to improve artificial insemination results in the cattle

  原山 洋, 中嶋 昭雄

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (C), Kobe University, Apr. 2022 - Mar. 2025


• Studies on expressions and roles of INSL3 secretory dynamics and its receptor expression in reproductive organs and germ cells in female cattle

  川手 憲俊, 西村 亮, 原山 洋, 古山 敬祐

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research, Grant-in-Aid for Scientific Research (C), Osaka Prefecture University, Apr. 2021 - Mar. 2025

  2021年度は、１）ウシの発情周期の主席卵胞と黄体および卵胞嚢腫のINSL3分泌能を調べることを目的とし、主席卵胞と嚢腫卵胞の卵胞液および黄体組織のINSL3と性ステロイド（テストステロン、エストラジオール-17β、プロジェステロン）量を解析するとともに、培養黄体細胞からのINSL3とプロジェステロンの分泌能を分析した。同時に、２）正常雌ウシと卵胞嚢腫牛の血中INSL3濃度の動態を解明するために、雌ウシの血中INSL3濃度を測定できるサンドイッチ時間分解蛍光免疫測定（TRFIA）法の開発に取り組んだ。 １）については、食肉処理場において雌ウシの卵巣を採取し、発情周期中の主席卵胞の卵胞液と黄体の黄体組織のINSL3と性ステロイド量を測定した。その結果、主席卵胞のINSL3分泌はその成熟にともなって増加することが示唆された。卵胞のテストステロンとエストラジオール-17β量は卵胞の嚢腫化により低下するが、INSL3量は増加することが示唆された。黄体組織のINSL3とプロジェステロン含量はその開花期に増加することが示された。一方、黄体から黄体細胞を分離して培養し、培養液のINSL3濃度を測定した結果、INSL3分泌能は開花期に上昇するが、その分泌は黄体形成ホルモン（LH）に依存しないことが示された。 ２）については、サンドイッチTRFIA法に必要な検出用の抗ウシINSL3ペプチド抗体を作成し、そのビオチン標識を実施した。さらに補足用ウシINSL3抗体、ウシINSL3標準品、ビオチン標識検出用抗体およびユーロピウム標識ストレプトアビジンを用いてサンドイッチTRFIAの測定法の諸条件の検討を行った。その結果、最小検出濃度が0.1 ng/mLの高感度測定法を確立することができた。雌ウシ血漿サンプルを本法で測定したところ、血中INSL3濃度を無抽出で測定できることが判明した。


• 融合因子SOFの機能解析を通した精子－卵子の細胞膜融合機構の解明

  野田 大地, 原山 洋

  日本学術振興会, 科学研究費助成事業, 基盤研究(B), Apr. 2020 - Mar. 2024

  sperm-oocyte fusion required (SOF)2～3を特異的に認識する抗体を得ることができなかったが、バックアッププランとして作製していたSOF2や3のC末端側にアフィニティータグを融合させたタンパク質を発現するトランスジェニックマウスを用いて、免疫染色法によりSOF2や3が先体反応後の精子頭部に局在することを明らかにできた。さらに、IP/MS解析によりSOF2や3と相互作用する因子の同定にも成功した。そこで、マウスの心臓、肝臓、脾臓、腎臓、脳、精巣、卵巣、および子宮を採取し、得られた各組織由来のcDNAを使ってPCRを行ったところ、これらの遺伝子は精巣や精巣上体で発現することが明らかになった。
  正常および低繁殖症を示す雄ウシの生殖細胞（精巣や成熟精子など）を採取して、ウェスタンブロティングを行ったところ、両者の間で差がある受精関連遺伝子を見出した。
  マウス既知融合関連遺伝子の機能が種を超えて保存されているかを調べる目的で、他動物種の融合関連遺伝子を発現するトランスジェニックマウスの作製に成功した。


• Analyses of 3-dimensional rotation for the purpose of reconsidering criteria for the evaluation of flagellar functions in bull sperm

  HARAYAMA HIROSHI

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Challenging Research (Exploratory), Challenging Research (Exploratory), Kobe University, Jun. 2019 - Mar. 2021, Principal investigator

  In this study regarding a new candidate "3-dimensional rotation (3D-R)" of the criterion for the evaluation of flagellar functions in bull cryopreserved sperm, we carried out experiments to elucidate the mechanism for its occurrence, to characterize it biologically and to examine its validity. The obtained results were listed below. (1) The cryopreservation-related occurrence of 3D-R is regulated by reduction of the activity of thapsigargin-sensitive Ca2+ pumps in the sperm neck and the consequent increases of the intracellular Ca2+ level. (2) The 3D-R is one of the typical patterns of motility which sperm exhibit during the capacitation process. (3) The 3D-R is a potentially criterial parameter for the evaluation of the flagellar functions in bull cryopreserved sperm.


• Studies on the mechanisms that cholesterol exerts its therapeutic effects on boar spermatozoa affected with summer infertility

  Murase Tetsuma

  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), Gifu University, Apr. 2017 - Mar. 2020, Coinvestigator

  cAMP content of boar spermatozoa stored up to 10 days with added cholesterol was not different among storage days. cAMP content in spermatozoa stored and incubated for stimulation of the acrosome reaction with the calcium ionophore A23187 slightly decreased and then rose during stimulation. Magnesium decreased the acrosome reaction triggered by A23187 while polymyxin B enhanced it. When frozen-thawed bull spermatozoa were used as a model, damaged acrosome, hyperactivation and the acrosome reaction were independent parameters from one another, suggesting that this should be taken into account when boar sperm function is investigated. Exposure of immature oocytes to adipose-derived stem cells (ADSC) and/or mechanical vibration during maturation in culture led to the increase in the sperm penetration and male pronuclear formation. A very effective method to collect, isolate and culture ADSC has been established in order to use them for increasing fertilizing ability of boar spermatozoa.


• Characterization and utilization of SPACA1 proteins for the purpose of establishment of a new method to predict freezability of bull sperm before cryopreservation

  HARAYAMA HIROSHI

  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, Apr. 2016 - Mar. 2019, Principal investigator

  The aims of the present study were to develop a new method to predict freezability of bull sperm before cryopreservation and to increase its reliability by disclosing molecular methodology. In this study, we succeeded in the development of the method to predict freezability of bull (Japanese Black bull) sperm before cryopreservation according to the criterion of the distribution of SPACA1 proteins. We also suggested that aberrant distribution of SPACA1 proteins is related to abnormal maturation of the sperm in the epididymis, and that low fertilizing ability of cryopreserved sperm with aberrant distribution of SPACA1 proteins is linked to poor results in the artificial insemination using them.


• Etiology of boar spermatozoa affected with summer infertility in an attempt to develop a novel therapy

  Murase Tetsuma, SHIMIZU Kenji, +HARAYAMA Hiroshi

  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), Gifu University, Apr. 2014 - Mar. 2017, Coinvestigator

  It was considered that one of the mechanisms underlying the hypersensitive induction of the acrosome reaction may be due to an increased content of cAMP [cAMP]i in boar spermatozoa collected during summer. Addition of cholesterol or removal of bicarbonate in the semen extender did not affect [cAMP]i of spermatozoa. Thus, a mechanism that cholesterol delays the induction of the acrosome reaction may be the effect on the generation of cAMP during the process of the acrosome reaction raher than during storage. Addition of magnesium lowered the occurrence of the acrosome reaction. The staining method of sperm acrosomes with FITC-PNA was modified. Preincubation of boar spermatozoa with adipose-derived stem cells (ADSC) increased the ability of in vitro fertilizing ability. Cholesterol, magnesium and ADSC are prospective in applying to a 'therapy of boar spermatozoa' suffering from summer infertility.


• Expression of claudin family proteins in preimplantation embryos in the pig

  MIYAKE MASASHI, HARAYAMA Hiroshi

  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, Apr. 2013 - Mar. 2016, Coinvestigator

  The expression of claudin family, tight junction (TJ) were examined in pig blastocysts derived from activated diploids. Blastocysts expressed both mRNAs and proteins of claudin-1, -2, -4, -6, -7, -8, -9, -12 and -14. While claudin-4, -6 and -7 localized along the cell-boundary TJ structure, the other claudins were cytoplasmic. These results suggest that claudin-4, -6 and -7 are important for the TJ functions in pig blastocysts. Knockdown of claudin-1 and -4 by their siRNA severely suppressed the early cleavage. To examine whether expression patterns of the claudins will be employed as markers for the potential developmental ability of in vitro produced blastocysts in future, features of glucose metabolism was examined in pig preimplantation embryos. It was shown that recommendation of carbohydrates are different depending on the preimplantation developmental stages and hexose biosynthesis pathway and GlcNAcylation have important roles in pig preimplantation development.


• Development of methods to evaluate molecular characteristics of livestock spermatozoa by the detection of aberrant fertility-regulating factors

  HARAYAMA HIROSHI, MURASE Tetsuma, FUKUSHIMA Moriyuki

  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, Apr. 2013 - Mar. 2016, Principal investigator

  The aim of the present study was to develop a new method to evaluate sperm molecular characteristics for identifying AI-subfertile males of livestock. In this study, we succeeded in the development of the method to evaluate molecular characteristics of bull (Japanese Black bull) cryopreserved and freshly ejaculated spermatozoa by the observation of acrosomal tyrosine-phosphorylated proteins. Moreover, we also showed that outcomes of AI and in vitro fertilization can be predicted by this evaluation for the spermatozoa.


• Enhancement of the tolerance to the storage in boar and mouse spermatozoa by the re-maturation in vitro

  HARAYAMA Hiroshi

  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, Kobe University, 2011 - 2012, Principal investigator

  There are great differences in the freezability of mammalian spermatozoa among species. In cattle, high freezability of the spermatozoa enables us to use frozen semen in the AI program. In this study, we indicated the molecular mechanism for epididymal maturation-related stabilization of the plasma membrane which brings high freezability to bull spermatozoa. In addition, we attempted to develop in vitro incubation system to enhance sperm freezability on the basis of the above-mentioned molecular mechanism.


• Development of clinical therapeutic methods against failure of fertilizing functions causing summer infertility in boars

  MURASE Tetsuma, HARAYAMA Hiroshi, MIYAKE Masashi

  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), Gifu University, 2009 - 2011, Coinvestigator

  In an attempt to improving boar sperm quality which decreases in summer and stable provision with semen leading to stable production of pork during the year, this study investigated a new diluent for boar semen storage, a new method to evaluate frozen-thawed bull sperm quality and development of a method to improve fertilizing ability of frozen-thawed bull spermatozoa. Bull spermatozoa were used as a reference to boar spermatozoa. Modified diluents for boar semen improved conception rates. In bulls, new method for sperm evaluation was found out and modification of diluent for freezing bull semen enabled improvement of sperm fertilizing ability.


• Targeted therapy for defect of fertilizing ability in the spermatozoa from subfertile livestock

  HARAYAMA Hiroshi, MIYAKE Masashi, MURASE Tetsuma, FUKUSHIMA Moriyuki

  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, 2008 - 2010, Principal investigator

  The initial step of animal reproduction is accomplishment of in vivo fertilization. For the purpose of successful fertilization, spermatozoa are required to show a precise process in the exertion of their fertilizing ability. In this study, we indicated that pivotal regulators of the intracellular signaling and glucose metabolism are potential targets to suppress precocious fertilization-related events in the spermatozoa. In addition, we also suggested that defective phosphorylation at the tyrosine residues of acrosomal proteins is one of causal factors for unstable structure in the spermatozoa from subfertile male animals.


• An invention of method to predict reproductive performance of Japanese Black cattle by the evaluation of sperm ability to exhibit flagellar hyperactivation

  HARAYAMA Hiroshi, MIYAKE Masashi, MURASE Tetsuma

  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, 2006 - 2007, Principal investigator

  1.Induction of sperm flagellar hyperactivation: The treatment with a cell-permeable analog cBiMPS (0.1 mM) for 3 h was effective for the induction of sperm flagellar hyperactivation. This reveals that flagellar hyperactivation is regulated via the cAMP singnaling in the sperm from Japanese Black cattle. 2. Characterization of the cAMP signaling: Our previous reports indicate that the cAMP-PKA-SYK signaling cascade is involved in the sperm flagellar hyperactivation in boars. However, this study has suggested that it is regulated via different cAMP signaling cascades in Japanese Black cattle. A possible downstream element of the cAMP signaling has been found as a focal adhesion kinase (FAK)-like protein. 3. Biological markers for the prediction of male reproductive performance: The treatment with cBiMPS rapidly induced transformation of motility pattern to curling movement in the sperm from normal bulls. However delayed transformation was observed in the sperm from subfertile bulls which were produced in Gifu Prefecture. On the other hand, relatively larger reduction of the motility was observed during the cBiMPS treatment in the sperm from subfertile bulls which were produced in Hyogo Prefecture. Namely, different responses of spermatozoa to cBiMPS may be valid as biological markers for prediction of male reproductive performance of Japanese Black cattle. 4. Biochemical markers for the prediction of male reproductive: performance: Detection levels of 130-kDa, 105-kDa and 94-kDa cAMP-dependently tyrosine-phosphorylated proteins were likely correlated with the percentages of hyperactivated sperm after the treatment with cBiMPS. Thus, these proteins are candidates of biochemical markers for the prediction of male reproductive performance of Japanese Black cattle. Additionally the 94-kDa tyrosine-phosphorylated protein has been identified as a FAK-like protein.


• An invention of method to suppress expression of sperm fertilizing ability for the purpose of establishment of production system of porcine embryos

  HARAYAMA Hiroshi, MIYAKE Masashi, MURASE Tetsuma

  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, 2004 - 2005, Principal investigator

  1.Identification of cAMP-dependent protein tyrosine kinase : A spleen tyrosine (Y) kinase (SYK) was activated via the cAMP-PKA signaling in the sperm connecting and principal pieces. The mechanism for SYK activation is quite different between spermatozoa and lymphocytes, suggesting that the activation of sperm SYK is controlled by a novel signaling system. 2.Observation of relationship between SYK activation and expression of sperm fertilizing ability : The cAMP analog-treated spermatozoa exhibited flagellar hyperactivation coincidently with the SYK activation. This indicates a role of SYK as a regulator for sperm fertilizing ability. 3.Analyses of cDNA sequence of testicular SYK : The sequence of PCR product (590 bps) amplified from testicular cDNA was same as that of pig spleen syk gene (the C-terminal catalytic domain of SYK), confirming production of SYK in the testis. 4.Identification and functional analyses of sperm SYK substrate : The SYK substrate was a phospholipase Cγ1 (PLCγ1). This enzyme, which was cAMP-dependently phosphorylated at the Y352 residue, bound to and activated PLCγ1 by the phosphorylation at the Y783 residue. Additionally, chemical inhibition of PLCγ1 with U-73122 was valid to the suppression of over-expression of flagellar hyperactivation. 5.Trial to introduce PLCγ1 peptide fragments into spermatozoa : Although two kinds of commercial kits were used in this experiment, we faced problems of methodology that unknown factors included in the reagents and introduction protocol severely damaged sperm motility. A possible method for peptide introduction remains to be improved. 6.Identification and functional analyses of other regulatory factors of sperm fertilizing ability : Sperm PKC was controlled via the cAMP-PKA signaling. The chemical inhibition of sperm PKC with Ro-32-0432 was valid to the suppression of over-expression of flagellar hyperactivation.


• Changes and functions of Anti-Agglutinin in boar spermatozoa during their capacitation

  KATO Seishiro, HARAYAMA Hiroshi

  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, 1999 - 2001, Coinvestigator

  Three following experiments have been undertaken in this study ; 1. changes of sperm-bound anti-agglutinin during the capacitation, 2 : isolation of anti-agglutinin from seminal plasma and functional characterization of this protein on the regulation of sperm capacitation, and 3 : signaling cascade regulating sperm head-to-head agglutination. Significant results have been obtained in Experiments 1 and 3 and summarized here. Experiment 1 has revealed changes in anti-agglutinin bound to spermatozoa during the incubation designed to promote the capacitation in vitro. The relative amount of sperm-bound anti-agglutinin, assessed by Western blotting-densitometric analyses, was almost halved after the first 45-min incubation and then decreased gradually thereafter. Immunocytocheinical observation showed that the decrease of the sperm-bound anti-agglutinin occurred mainly on acrosomes in many spermatozoa during the incubation. These results indicate that a large portion of the anti-agglutinin bound to sperm acrosomes is released at an early stage of the Capacitation process in vitro. Experiment 3 has been conducted to reveal roles of bicarbonate-adenylyl cyclase (AC)-cAMP-protein kinase (PK) system in sperm head-to-head agglutination. In the samples incubated in Ca-free mKRB supplemented with 100 μM forskolin (an AC activator), significantly higher percentages of the spermatozoa were agglutinated with one another at the acrosome, as compared with control samples. Addition of dibutyryl CAMP (dbcAMP, an activator of CAMP-dependent PK : PKA), instead of forskolin, increased the percentages of head-to-head agglutinated spermatozoa in a dose-dependent manner between 1 and 1,000 μM. However, the effects of the addition of dbcAMP (1,000 μM) were attenuated by treatments with Rp-cAMPS (1,000 μM, a PKA inhibitor) or H-89 (5 μM, a PKA inhibitor). These results indicate that bicarbonate-AC-cAMP-PK system mediates a signaling pathway leading to head-to-head agglutination of spermatozoa.


• ブタ精子のキャパシテーション過程での頭部間凝集に及ぼすカルシウムの影響

  原山 洋

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Encouragement of Young Scientists (A), Grant-in-Aid for Encouragement of Young Scientists (A), Kobe University, 1999 - 2000, Principal investigator

  実験1.カルモジュリン(CaM)の役割:既報(Chan and Dukelow,1985;Leclerc et al.,1990,1992)によると,精子におけるCaM濃度およびCaMの特異タンパク質への結合性はキャパシテーションとともに低下することから,CaMはキャパシテーションに抑制的に作用していると推察されている.そこで,本研究では成分からキャパシテーション誘起因子を除去した媒液中で精子をインキュベートした際のCaM阻害剤の影響について検討した.予備実験においてCalmidazoliumとTrifluoperazineを用いたが,これらの薬剤は精子に対する毒性が強く,本実験での使用には不適切であった.そこで,比較的毒性の低かったW7(5〜10μM)を用いたところ,頭部間凝集精子率は著しく上昇した.この結果から,CaMは精子の頭部間凝集に抑制的に作用していると考えられる. 実験2.電位依存性カルシウムチャンネル(VDCCs)の役割:昨年度の研究において10〜100μMの濃度では精子の頭部間凝集に有意な抑制作用を示さなかったL型VDCCs阻害剤のNifedipineとNitrendipineをより高い濃度(1mM)で添加したところ,頭部間凝集率は有意に低下した.他方,T型VDCCsの役割については,予備実験で精子に対する毒性が比較的少ないことが判明した阻害剤のPimozideとAmiloride(いずれも0.1〜10μM)を用いて検討したが,いずれも凝集精子率に有意な変動は認められなかった.以上の結果から,精子の頭部間凝集の発生においてVDCCsが機能している可能性が示唆されたが,L型阻害剤で抑制効果が認められた濃度は1mMと高く,またT型阻害剤(0.1〜10μM)には効果が認められなかったことから,VDCCsの種類を特定するには至っていない.


• ブタ活性化卵子の初期発生能ならびに単為発生胚の初期胚特異タンパク質の発現性

  MIYAKE Masashi, KUREBAYASHI Satoshi, HARAYAMA Hiroshi

  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 - 1999, Coinvestigator

  This study was designed to establish serum-free culture system for porcine early embryos and to examine the developmental limitation of parthenogenetic porcine diploids. In vitro matured porcine oocytes were electro-stimulated for activation and were treated with cytochalasin B. These oocytes were cultured for 168 hours in various simple media that were based on modified Whitten' medium with 0.5mg/ml hyaluronic acid (mWM). Osmolarity of media, concentration of polyvinyl alcohol (PVA) as substitutes for ovine serum albumin (BSA) and effects of amino acids were examined. These studies clarified following items ; 1.Diploids cultured in isotonic mWM (309mOsmol) throughout culture period resulted in very low frequency (a few %) of development to the blastocyst stage by the failure of compaction and formation of blastocoel. Fourteen to 20% of diploids cultured in hypotonic mWM (254 mOsmol) developed to the blastocyst stage. The cause of the osmotic effect is just osmolarity of a media, but Na+/K+ ratio. 2.Isotonic condition (280 to 320 mOsmol) is more beneficial for the culture of diploids up to 48 hours after activation, and hypotonic condition (220 to 270 mOsmol) is better for the development of diploids from 72 hours after activation. 3.More than 0.5 mg/ml of PVA as substitutes for BSA supports well the development of diploids to the blastocyst stage, but their expansion. The addition of amino acids solution (AA) mixed essential (EA) and nonessential amino acids (NEA) to PVA-mWM from the biginning of culture causes strong 4-cell blocking of diploids. The frequency to the blastocyst stage is significantly lower than AA-free condition. 5.The addition of EA for 0 to 48 hours after activation brings sever 4-cell blocking to the diploids. The blocking effect is weakened by the addition of NEA, and NEA can support the development of diploids to the blastocyst stage as like as BSA. 6.The addition of EA 48 after activation well supports their development to the blastocyst stage as like as BSA. Moreover, most of blastocysts cultured under the condition show the hatching process. To determine the in vivo developmental of parthenogenetic diploids, activated oocytes at the 3- to 4-cell stage 48 hours after activation were transferred to recipients. The animals were sacrificed on 18 to 30 days after activation, and the number of fetuses and existence of corpus lutea was examined. 1.In one of four trails, 30-days fetuses were obtained (35%), but in the other 3 cases no fetuses was obtained. Aborted fetuses were observed on 28- to 29-day after activation in one animal. These results suggest that the developmental limitation of parthenogenetic diploids is around 30 day after activation. 2.Parthenogenetic fetuses on 25-day, at the early stage of post implantation, were obtained in many trials, indicating that the developmental ability of diploids to the stage was relatively high (41% of transferred eggs), however, cyst-like structure of the primitive heart and liver, and acephalus were observed in some of fetuses collected around 20-day. 3.NMR images shows that some of fetuses with normal appearance have abnormal cavity formation in the brain. Expression of some of imprinted genes that have been clarified their imprinting in the mouse and/or human in parthenogenetic porcine diploids was also examined in the present study. 1.As the sequence of most of the imprinted genes in the pig, probes for porcine genes were designed from the sequence of mouse and human genes. PCR products were obtained from normal porcine fetuses using probes for p57KIP2, PEG1/MEST, SNRPN and H19. 2.Expression of candidates of imprinted genes were compared between normal and parthenogenetic fetuses by northern-hybridization using PCR products of IGF2, IGF2R and WT1 in addition to the above PCR products and sub quantitative PCR. The expression of IGF2 is stronger in normal fetuses than parthenogenotes, and that of p57KIP2 was almost same between them.


• Studies on production of pig embryos from growing oocytes

  KATO Seishiro, MIYANO Takashi, MIYAKE Masashi, KANNAN Yasuyuki, TOMINAGA Keiichiro, HARAYAMA Hiroshi

  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 - 1999, Coinvestigator

  Several hundred thausand primordial follicles are contained in the ovaries of domestic animals. The development of in vitro culture systems for the growth of small oocytes in primordial or small follicles is expected to provide a new source of a large population of oocytes for livestock production. This study was conducted to develop Culture systems for small oocytes from pig ovaries. Preantral follicles containing oocytes with various sizes were isolated from pig ovaries and cultured in collagen gel for up to 16 days in the presence of serum, FSH and oestradiol. Waymouth MB7 52/1 was used as a basic culture medium for oocyte growth. After culture, the oocytes enclosed by one or two layers of cumulus cells were cultured for meiotic maturation and inseminated with spermatozoa. The results obtained were as follows. 1. Pig growing oocytes grew to their final size, acquired meiotic competence du ring in vitro culture, and were penetrated by spcermatozoa in vitro, although there was a very low level of success in fertilization. In contrast to the growing oocytes, no culture methods were available for much smaller oocytes. We tried to culture primordial follicles which contained small oocytes 30 μm in diameter, but the integrity of the follicles was soon broken and the follicles survived for only a few days. 2. Using culture systems, we demonstrated that granulosa cells regulate oocyte growth, and that antrum formation of granulosa cells was induced only by meiotic-competent oocytes, indicating that the oocytes secrete a substance(s) that induces differentiation of granulosa cells. 3. Of four glycosaminoglycans examined, hyalronic acids and chondroitin sulfate A supported the development of in vitro-matured and -fertilized pig oocytes to the blastocyst stage. 4. In addition, degradation of cyclin B1 molecules and MAP kinase deph osphorylation during fertilization, localisation of phosphorylated MAP kinase during the transition from meiosis I to meiosis II, assembly and transformation of the spindle during the progression through meiotic cell cycle, and the level and localization of CENP-E and kinetochore numaber during maturation were examined.


• Anti-Agglutinin出現機序の生化学的および免疫組織学的解析

  原山 洋

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Encouragement of Young Scientists (A), Grant-in-Aid for Encouragement of Young Scientists (A), Kobe University, 1997 - 1998, Principal investigator

  3頭の成熟雄ブタから採取した精巣上体を頭2部位,体2部位および尾1部位の合計5部位に区分し,パラホルムアルデヒド固定,凍結およびクリオスタットによる薄切の後に各部位でのAnti-Agglutininの分布を免疫組織学的に観察した。その結果,はじめてAnti-Agglutininが検出されたのは精巣上体の体近位部で,精巣上体管の上皮と管腔内容物の両方において認められた。また,精巣上体体遠位部においてもほぼ同様の検出結果が得られた。さらに,精巣上体尾の管腔内容物においてもAnti-Agglutininは検出されたが,管上皮にはほとんど認められなかった。他方,3頭の成熟雄ブタから12種類の臓器(精巣上体,精巣,精のう腺,前立腺,心臓,肝臓,腎臓,脾臓,胃,小腸,肺および筋肉)を採取し,臓器組織の一部を1mMAPMSF,1mMEDTAおよび1%TritonX-100添加リン酸緩衝生理食塩水中でホモジネートにした後,そのホモジネートを用いてSDSポリアクリルアミドゲル電気泳動およびウエスタンブロッティングを行ったところ,Anti-Agglutininが検出されたのは精巣上体のみで,中でも精巣上体体において最も強い反応が得られた。以上の結果から,精巣上体においてAnti-Agglutininを生産および分泌する部位は体であると考えられる。なお,精巣上体管腔内に分泌されたAnti-Agglutininの一部は精子の先体と結合することを細胞免疫学的手法により合わせて明らかにした。


• 精巣上体液中の精子頭部間凝集抑制タンパク質を修飾する糖鎖の特性解析

  原山 洋

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Encouragement of Young Scientists (A), Grant-in-Aid for Encouragement of Young Scientists (A), Kobe University, 1996 - 1996, Principal investigator

  1.研究実績の概要:哺乳動物の精巣上体における精子の成熟変化のひとつである頭部間凝集性低下の分子機構を明らかにする目的で、精子頭部間凝集抑制タンパク質(Anti-Agglutinin:以下AAとする)を修飾する糖鎖の特性を解析し、以下の結果を得た。 2.糖鎖修飾の違いがAAの精子への結合性に及ぼす影響:AAにおいて、分子不均一性が精子への結合性に大きく影響するが、この分子不均一性の原因を明らかにする目的で、精巣上体尾液(30μg)を尿素存在下で2次元電気泳動した後、レクチンブロッティング法に従ってコムギ胚芽凝集素(WGA)との反応性について調べた。その結果、精子との結合性を備える等電点5.8のスポットはWGAと反応したが、精子と結合しない等電点5.6および5.95のスポットは反応しなかった。従って、糖鎖のシアル酸化の有無が分子不均一性の一因であり、また精子への結合性に影響すると考えられる。 3.修飾糖鎖の分子量測定:AAからGlycoFree-Kitを用いて糖鎖を完全に除去し、その際の分子量の変化をMALDI-TOF-Mass Spectrometric Analysisにより測定したところ、無処理のAAは分子量18900-19600においてブロードしたピークとして検出されたが、糖鎖除去後の試料は最高700小さい分子量においてシャープやピークを示した。このことから、総分子量で700以下の糖鎖がAAを修飾しており、その修飾糖鎖は多種類存在すると考えられる。 4.脱シアル酸がAAの生理活性に及ぼす影響:ノイラミニダーゼ(0.5U/ml)を用いて脱シアル酸化したAAを洗浄精子浮遊液に添加し37℃で5時間インキュベートしたところ、AAの精子凝集抑制作用は若干低下する傾向を示した。