哺乳類の受精における卵と精子の相互作用に関する研究

• 研究課題/領域番号: 09044240
• 研究種目: 基盤研究(B)
• 配分区分: 補助金
• 応募区分: 一般
• 研究分野: 動物生理・代謝
• 研究機関: 岡崎国立共同研究機構
• 研究代表者: 毛利 秀雄 岡崎国立共同研究機構, 基礎生物学研究所, 所長 (70012268)
• 研究分担者: 岡部 勝 大阪大学, 遺伝情報実験施設, 教授 (30089875) ; 石島 純夫 東京工業大学, 理学部, 助手 (70193315) ; 奥野 誠 東京大学, 大学院・総合文化研究科, 助教授 (40143325) ; 小川 和男 岡崎国立共同研究機構, 基礎生物学研究所, 助教授 (30132731) ; YANAGIMACHI R. DEP. OF ANATOMY & REPROD. IO., UNIV. OF HAWAII, PORFESSOR ; SAVAGE J.s. DEP. BIOL., UNIV. NEW ORLEANS, ASSOCIATE PROFESSOR ; SUAREZ S.s. DEP. BIOMEDICAL SCI., CORNELL UNIV., PROFESSOR ; STEWART・SAVA ジェイ ニューオリンズ大学, 生物科学, 副教授 ; SUAREZ S.S. コーネル大学, 獣医学部, 副教授 ; YANAGIMACHI アール ハワイ大学, 医学部, 教授 ; SAVAGE J.STE ニューオリンズ大学, 生物科学, 副教授
• 研究期間 (年度): 1997 – 1999
• 研究課題ステータス: 完了 (1999年度)
• 配分額: 9,400千円 (直接経費: 9,400千円); 1999年度: 2,100千円 (直接経費: 2,100千円); 1998年度: 3,200千円 (直接経費: 3,200千円); 1997年度: 4,100千円 (直接経費: 4,100千円)
• キーワード: 哺乳類精子 / 超活性化 / 受精能獲得 / 鞭毛運動 / 先体反応 / 卵・精子相互作用 / 透明帯通過

研究概要

本研究班はそれぞれのメンバーが随時互いの研究室を訪れて共同研究を行ってきた。この3年間に得られた結果は以下の通りである。(1)本研究の中心課題であった精子の超活性化について、ハムスター、モルモット、ウシ、サルを用いて色々な面から検討が加えられてきた。高速度ビデオ装置等による鞭毛波形の観察から、超活性化をおこすと中片部が大きく曲がるようになって非対称的な運送を示し、さらに先体反応をおこすと一層曲がりやすく振幅も大きくなるが対称的な運動になることが分かった。また精子先体に緑色蛍光を発するGFPタンパク質を発現する形質転換マウスを利用してこの間の経過を追った。(2)超活性化はハムスターやウシではプロカインによって、またモルモットではリソレシチンによって誘起されることが分かった。(3)精子の運動状態には停止状態、弱い活性化状態、強い活性化状態、超活性化状態があり、それぞれに特異的なタンパク質がリン酸化されることが明らかになった。(4)超活性化の初期にはカルシウムの働きで非対称性の波が起こる。またcAMPによって中片部の対称的な振幅の大木運動がひきおこされる。カルシウムの貯蔵場所は鞭毛基部の特殊な構造であることが示されたが、遊離したカルシウムはミトコンドリアの活性を高めて超活性化を支えていると思われる。(5)卵透明帯を通過しつつある精子の鞭毛は対称な振幅の大きな屈曲を作っていることが観察され、このような運動によって精子が透明帯を通過するための充分な推力が与えられることが明らかになった。(6)運動活性化過程におけるグルコースの役割が解析され、また精子鞭毛運動のモーターであるダイニンの軽鎖2本がTctex遺伝子に相当することが示された。(7)この他、精子や精子形成細胞の卵細胞質内注入によるマウスの作成、死体から得られたマウス精子の受精能についての研究などが行われた。

研究成果

• [文献書誌] Kubo-Irie ,M. et al.: "Morphological evidence that spermatogenetic cells undergo apoptosis within testis of diapausing swallowtail pupa(Astrophaneura alcinous)"J.Exp.Zool.. 283. 194-201 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Mohri, H. et al.: "Characterization of outer arm dynein in sea anemone, Anthopleura midori"Cell Motil, Cytoskel.. 44. 202-208 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Mohri, H.: "Tubulin 30 years later"Cell Struct. Funct.. 24. 265-267 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Si, Y. and Okuno, M.: "Regulation of microtublue sliding by 36-kDa phosphoprotein in Hamster sperm flagella"Mol. Reprod. Dev.. 52. 328-334 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Si, Y. and Okuno, M.: "Role of tyrosine phosphorylation of flagellar proteins in hamster sperm hyperactivation"Biol. Reprod. 61. 240-246 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] 石島純夫: "精子運動のメカニズム"日本受精着床学会雑誌. 14. 7-10 (1997)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Ishijima, S. et al.: "Comparative studies on the spermatozoan motility of Japanese fishes"Bull. Ocean Res. Inst. Univ. Tokyo. 33. 139-152 (1998)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Ishijima, S. et al.: "Movement of Turritella spermatozoa : direction of propagation and chirality of flagellar bends"Cell Motil. Cytoskel.. 44. 85-95 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Watanabe, S. et al.: "Mitochondrial thioredoxin reductase in bovine adrenal cortex : its purification, properties, nucleotide/amino acid sequences, and identification of selenocysteine"Eur. J. Biochem.. 204. 74-84 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Inaba, K. et al.: "Tctex2-related outer arm dynein light chain is phosphorylated at activation of sperm motility"Biochem. Biophys. Res. Commum. 256. 177-183 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Miyado, K. et al.: "Requirement of CD9 on the egg plasma membrane for fertilization"Science. 287. 321-324 (2000)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Hamada, Y. et al.: "Mutation in ankyrin repeats of the mouse notch2 gene induces early embryonic lethality"Development. 126. 7-10 (1997)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Nakanishi, T. et al.(Okabe, M.): "Real-time Observation of acrosomal dispersal from mouse sperm using GFP as a marker protein"FEBS Lett. 449. 277-283 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Matsumoto,A. et al.(Okabe, M.): "Suppression of STAT5 function in liver,mammary glands, and T cells in cytokine-inducible SH2-containing protein 1 transgenic mice"Mol. Cell Biol. 19. 6396-6407 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Kato, M. et al.: "Efficient selection of transgenic mouse embryos using EGFP as a marker gene"Mol. reprod. Dev.. 54. 43-48 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Perry, A. C. et al.: "Mammalian transgenesis by intracytoplamic sperm injection"Science. 284. 1180-1183 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Usui, N. et al.: "Ultrastructural changes in hamster spermatogenic cell nuclei after incorporation into homologous oocytes by electrofusion"Mole. Reprod. Dev.. 52. 66-73 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Perry, A. C. F. et al.: "A novel trans-omplementation assay suggests full mammalian oocyte activation is coordinated by multiple submembrane sperm components"Biol. Reprod. 60. 747-755 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Yanagida, K. et al.: "Successful fertilization and pregnancy following ICSI and electrical oocyte activation"Human Reprod. 14. 1307-1311 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Kishikawa, H., Tateno, H., and Yanagimachi, R.: "Fertility of mouse spermatozoa from cadavers maintained at 4^○C"J. Reprod. Feril.. 116. 217-222 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Cummins, J. M. et al.: "Fate of genetically marked mitochondrial DNA from apermatocytes microinjected into mouse zygotes"Zygote. 7. 151-156 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Kasai, T., Hoshi, K and R. Yanagimachi: "Effect of sperm immobilisation and demembranation on the oocyte activation rate in the mouse"Zygote. 7. 187-193 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Ho, H-C, Tnag C-Y, and Suarez, S. S.: "Three-dimensional structure of the Golgi apratus in mouse spermatids : a scanning electron microscope study"Anat. Rec.. 256. 189-194 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Revah, I. et al.: "The physiological state of bull sperm affects fucose and mannose binding properties"Bio. Reprod.. 62(in press). (2000)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Visconti, P. E. et al.(Stewart-Savage, J.): "Roles of bicarbonate, cAMP and protein tyrosine phosphorylation on capacitating and the spontaneous acrosome reaction of hamster sperm"Bio. Reprod.. 61. 76-84 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Stewart-Savage, J. et al.: "Developmental Basis of phenotypic variation in egg production in a colonial ascidian : primary oocyte production versus oocyte development"Bio. Bull.. 196. 63-69 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Suarez,S.S.: "The Male Gemete : From basic sience clinical applications."Cache Riever Press. 10 (1999)
  • 説明: 「研究成果報告書概要(和文)」より
• [文献書誌] Kubo-Irie, M. et al. (Mohri, H): "Morphological evidence that spermatogenic cells undergo apoptosis within testis of diapausing swallowtail pupa (Atrophaneura alcinous)."J. Exp. Zool. 283. 194-201 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Mohri, H. et al.: "Characterization of outer arm dynein in sea anemone, Antholeura midori."Cell Motil. Cytoskel. 44. 202-208 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Mohri, H.: "Tubulin 30 years later"Cell Struct. Funct.. 24. 265-267 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Si, Y. and Okuno, M.: "Regulation of microtubule sliding by 36-kDa phosphoprotein in Hamster sperm flagella."Mol, Reprod. Dev. 52. 328-334 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Si, Y and Okuno, M: "Role of tyrosine phosphorylation of flagellar proteins in hamster sperm hyperactivation."Biol. Reprod. 61. 240-246 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Ishijima, S. et al.: "Comparative studies on the spermatozoan motility of Japanese fishes."Bull. Ocean Res. Inst. Univ. Tokyo. 33. 139-152 (1998)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Ishijima, S. et al.: "Movement of turritella spermatozoa. Direction of propagation and chirality of flagellar bends."Cell Motil. Cytoskel.. 44. 85-95 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Watabe, S. et al. (Ogawa, K.): "Mitochondrial thioredoxin reductase in bovine adrenal cortex: its purification, properties, nucleotide/amino acid sequences, and identification of selenocysteine"Eur. J. Biochem.. 204. 74-84 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Inaba, K. et al. (Ogawa, K.): "Tetex2-related outer arm dynein light chain is phosphory-lated at activation of sperm motility."Biochem, Biophys. Res. Commun.. 256. 177-183 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Miyado, K. et al. (Okabe, M.): "Requirement of CD9 on the egg plasma membrane for fertilization."Science. 287. 321-324 (2000)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Hamada, Y. et al. (Okabe, M.): "Mutation in ankyrin repeats of the mouse noich2 gene induces early embryonic lethality."Development. 126. 7-10 (1997)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Nakanishi, T. et al. (Okabe, M.): "Real-time observation of acrosomal dispersal from mouse sperm using GFP as a marker protein."FEBS Lett. 449. 277-283 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Matsumoto, A. et al. (Okabe, M.): "Suppression of STAT5 functions in liver, mammary glands, and T cells in cytokine-inducible SH2-containing protein 1 transgenic mice."Mol, Cell Biol.. 19. 6396-6407 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Kato, M. et al. (Okabe, M.): "Efficient selection of transgenic mouse embryos using EGFP as a marker gene."Mol. Reprod. Dev.. 54. 43-48 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Perry, A. C. et al. (Okabe, M.): "Mammalian transgenesis by intracytoplasmic sperm injection."Science.. 284. 1180-1183 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Usui, N. et al. (Yanagimachi, R.): "Ultrastructural changes in hamster spermatogenic cell nuclei after incorporation into homologous oocytes by electrofusion."Mole. Reprod. Dev.. 52. 66-73 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Perry, A. C. F. et al. (Yanagimachi, R.): "A novel trans-complementation assay suggests full mammalian oocyte activation is coordinated by multiple submembrane sperm components."Biol. Reprod.. 60. 747-755 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Yanagida, K. et al. (Yanagimachi, R.): "Successful fertilization and pregnancy following ICSI and electrical oocyte activation."Human Reprod.. 14. 1307-1311 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Kishikawa, H., Tateno, H. and Yanagimachi, R.: "Fertility of mouse spermatozoa from cadavers maintained at 4 ℃."J. Reprod. Feril.. 116. 217-222 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Cummins, J. M. et al. (Yanagimachi, R.): "Fate of genetically marked mitochondrial DNA from spermatocytes microinjected into mouse zygotes."Zygote. 7. 151-156 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Kasai, T., Hoshi, K. and Yanagimachi, R.: "Effect of sperm immobilisation and demembranation on the oocyte activation rate in the mouse."Zygote. 7. 187-193 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Ho, H-C, Tang, C-Y and Suarez, S. S.: "Three-dimensional structure of the Golgi apratus in mouse spermatids: a scanning electron microscope study."Anat. Rec.. 256. 189-194 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Revah, I. Et al. (Suarez, S. S.): "The physiological state of bull sperm affects fucose and mannose binding properties."Bio. Reprod.. 62(in press). (2000)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Visconti, P.E. et al. (Stewart-Savage, J.): "Roles of bicarbonate, cAMP and protein tyrosine phosphorylation on capacitation and the spontaneous acrosome reaction of hamster sperm."Bio. Reprod.. 61. 76-84 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Stewart-Savage, J. et al.: "Developmental basis of phenotypic variation on egg production in a colonial ascidian : primary oocyte production versus oocyte development."Biol. Bull.. 196. 63-69 (1999)
  • 説明: 「研究成果報告書概要(欧文)」より
• [文献書誌] Kubo-Irie, M. et al. (Mohri H.): "Morphological evidence that spermatogenic cells undergo apoptosis within testis of diapausing swallowtail pupa (Atrophaneura alcinous)"J. Exp. Zool.. 283. 194-201 (1999)
• [文献書誌] Mohri, H. et al.: "Characterization of outer arm dynein in sea anemone, Anthopleura midori"Cell Motil. Cytoskel. 44. 202-208 (1999)
• [文献書誌] Mohri, H.: "Tubulin 30 years later"Cell Struct. Funct.. 24. 265-267 (1999)
• [文献書誌] Si, Y. and Okuno, M.: "Regulation of microtubule sliding by 36-kDa phosphoprotein in Hamster sperm flagella"Mol. Reprod. Dev.. 52. 328-334 (1999)
• [文献書誌] Si, Y. and Okuno, M.: "Role of tyrosine phosphorylation of flagellar proteins in hamster sperm hyperactivation"Biol. Reprod.. 61. 240-246 (1999)
• [文献書誌] 石島純夫: "精子運動のメカニズム"日本受精着床学会雑誌. 14. 7-10 (1997)
• [文献書誌] Ishijima, S. et al.: "Comparative studies on the spermatozoan motility of Japanese fishes"Bull. Ocean Res. Inst. Univ, Tokyo. 33. 139-152 (1998)
• [文献書誌] Ishijima, S. et al.: "Movement of Turritella spermatozoa : direction of propagation and chirality of flagellar bends"Cell Motil. Cytoskel. 44. 85-95 (1999)
• [文献書誌] Watabe, S. et al. (Ogawa K.): "Mitochondrial thioredoxin reductase in bovine adrenal cortex : its purification, properties, nucleotide/amino acid sequences, and idenification of selenocysteine"Eur. J. Biochem.. 204. 74-84 (1999)
• [文献書誌] Inaba, K. et al. (Ogawa,K.): "Tctex2-related outer arm dynein light chain is phosphorylated at activation of sperm motility"Biochem. Biophys. Res. Commun.. 256. 177-183 (1999)
• [文献書誌] Miyado, K. et al. (Okabe M.): "Requirement of CD9 on the egg plasma membrane for fertilization"Science. 287. 321-324 (2000)
• [文献書誌] Hamada, Y. et al.: "Mutation in ankyrin repeats of the mouse notch2 gene induces early embryonic lethality"Development. 126. 7-10 (1997)
• [文献書誌] Nakanishi, T. et al. (Okabe M.): "Real-time observation of acrosomal dispersal from mouse sperm using GFP as a marker protein"FEBS Lett. 449. 277-283 (1999)
• [文献書誌] Matsumoto, A. et al. (Okabe M.): "Suppression of STAT5 functions in liver, mammary glands, and T cells in cytokine-inducible SH2-containing protein 1 transgenic mice"Mol. Cell Biol.. 19. 6396-6407 (1999)
• [文献書誌] Kato, M. et al. (Okabe M.): "Efficient selection of transgenic mouse embryos using EGFP as a marker gene"Mol. Reprod. Dev.. 54. 43-48 (1999)
• [文献書誌] Perry, A.C. et al. (Okabe M.): "Mammalian transgenesis by intracytoplasmic sperm injection"Science. 284. 1180-1183 (1999)
• [文献書誌] Usui, N. et al. (Yanagimachi R.): "Ultrastructural changes in hamster spermatogenic cell nuclei after incorporation into homologous oocytes by electrofusion"Mole. Reprod. Dev.. 52. 66-73 (1999)
• [文献書誌] Perry, A.C.F. et al. (Yanagimachi,R.): "A novel trans-complementation assay suggests full mammalian oocyte activation is coordinated by multiple submembrane sperm components"Biol. Reprod. 60. 747-755 (1999)
• [文献書誌] Yanagida, K. et al. (Yanagimachi R.): "Successful fertilization and pregnancy following ICSI and electrical oocyte activation"Human Reprod. 14. 1307-1311 (1999)
• [文献書誌] Kishikawa, H., Tateno, H. and Yanagimachi, R.: "Fertility of mouse spermatozoa from cadavers maintained at 4℃"J. Reprod. Feril. 116. 217-222 (1999)
• [文献書誌] Cummins, J.M. et al. (Yanagimachi, R.): "Fate of genetically marked mitochondrial DNA from spermatocytes microinjected into mouse zygotes"Zygote. 7. 151-156 (1999)
• [文献書誌] Kasai, T., Hoshi, K and R. Yanagimachi: "Effect of sperm immobilisation and demembranation on the oocyte activation rate in the mouse"Zygote. 7. 187-193 (1999)
• [文献書誌] Ho, H-C, Tnag C-Y, and Suarez, S.S.: "Three-dimensional structure of the Golgi apratus in mouse spermatids ; a scanning electron microscope study"Anat. Rec.. 256. 189-194 (1999)
• [文献書誌] Revah, I. Et al. (Suarez, S.S.): "The physiological state of bull sperm affects fucose and mannose binding properties"Biol. Reprod. 62(in press). (2000)
• [文献書誌] Visconti, P.E. et al. (Stewart-Savage, J.): "Roles of bicarbonate, cAMP and protein tyrosine phosphorylation on capacitation and the spontaneous acrosome reaction of hamster sperm"Biol. Reprod. 61. 76-84 (1999)
• [文献書誌] Stewart-Savage J. et al.: "Developmental basis of phenotypic variation in egg production in a colonial ascidian : primary oocyte production versus oocyte development"Biol. Bull. 196. 63-69 (1999)
• [文献書誌] Suarez, S.S.: "The Male Gemete : From basic science to clinical applications"Cache Riever Press. 10 (1999)
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