1999 Fiscal Year Annual Research Report

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

Research Project

Project/Area Number	09044240
Research Institution	OKAZAKI NATIONAL RESEARCH INSTITUTE
Principal Investigator	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)
Co-Investigator(Kenkyū-buntansha)	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)
Keywords	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
Research Abstract	(1)精子の超活性化に関しては、ハムスターでの高速度ビデオ装置による記録から、受精能を獲得して超活性化すると中片部が大きく曲がるようになり、さらに先体反応をおこすと一層曲がりやすく大きな振幅を示すようになることが分った。また屈曲の仕方も非対称的な運動から対称的な運動へと変化した。しかし蛍光タンパクGFPを先体に発現させたマウスで観察したところでは、先体反応の前後でこのような変化はみられていない。超活性化はハムスターおよびウシでプロカインによって誘起された。(2)精子の運動の活性化については、停止状態、弱い活性化状態、強い活性化状態、超活性化状態に応じてそれぞれに特異的なタンパク質のリン酸化がおこることが明らかになった。超活性化では80kDaのタンパク質がリン酸化され、このタンパク質は鞭毛主部に局在する。(3)超活性化の初期ではカルシウムの働きによって非対称の波形が観察される。またcAMPによって中片部の対称な大きな振幅の運動がひきおこされる。カルシウムの精子内の貯蔵場所は鞭毛の基部にある特殊な構造であることがハムスターやウシの精子で明らかになった。(4)遊離したカルシウムはミトコンドリアに働きATP生産を高めて超活性化を支えると考えられる。(5)卵の透明帯を通過しつつある精子の運動の観察から、鞭毛は対称な振幅の大きな屈曲を作りながら通過していることが分かった。このような運動により精子が透明帯を通過するのに必要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の解析や、鞭毛のモータータンパク質であるダイニンのATPアーゼ活性部位の同定が行われた。(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。

Research Products

(27 results)

All Other

All Publications (27 results)

[Publications] 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)
[Publications] Mohri, H. et al.: "Characterization of outer arm dynein in sea anemone, Anthopleura midori"Cell Motil. Cytoskel. 44. 202-208 (1999)
[Publications] Mohri, H.: "Tubulin 30 years later"Cell Struct. Funct.. 24. 265-267 (1999)
[Publications] Si, Y. and Okuno, M.: "Regulation of microtubule sliding by 36-kDa phosphoprotein in Hamster sperm flagella"Mol. Reprod. Dev.. 52. 328-334 (1999)
[Publications] Si, Y. and Okuno, M.: "Role of tyrosine phosphorylation of flagellar proteins in hamster sperm hyperactivation"Biol. Reprod.. 61. 240-246 (1999)
[Publications] 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)
[Publications] Ishijima, S. et al.: "Comparative studies on the spermatozoan motility of Japanese fishes"Bull. Ocean Res. Inst. Univ, Tokyo. 33. 139-152 (1998)
[Publications] Ishijima, S. et al.: "Movement of Turritella spermatozoa : direction of propagation and chirality of flagellar bends"Cell Motil. Cytoskel. 44. 85-95 (1999)
[Publications] 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)
[Publications] 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)
[Publications] Miyado, K. et al. (Okabe M.): "Requirement of CD9 on the egg plasma membrane for fertilization"Science. 287. 321-324 (2000)
[Publications] Hamada, Y. et al.: "Mutation in ankyrin repeats of the mouse notch2 gene induces early embryonic lethality"Development. 126. 7-10 (1997)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] Perry, A.C. et al. (Okabe M.): "Mammalian transgenesis by intracytoplasmic sperm injection"Science. 284. 1180-1183 (1999)
[Publications] 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)
[Publications] 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)
[Publications] Yanagida, K. et al. (Yanagimachi R.): "Successful fertilization and pregnancy following ICSI and electrical oocyte activation"Human Reprod. 14. 1307-1311 (1999)
[Publications] Kishikawa, H., Tateno, H. and Yanagimachi, R.: "Fertility of mouse spermatozoa from cadavers maintained at 4℃"J. Reprod. Feril. 116. 217-222 (1999)
[Publications] Cummins, J.M. et al. (Yanagimachi, R.): "Fate of genetically marked mitochondrial DNA from spermatocytes microinjected into mouse zygotes"Zygote. 7. 151-156 (1999)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] 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)
[Publications] Suarez, S.S.: "The Male Gemete : From basic science to clinical applications"Cache Riever Press. 10 (1999)

1999 Fiscal Year Annual Research Report

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

Principal Investigator

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)

Research Products

[Publications] 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)

[Publications] Mohri, H. et al.: "Characterization of outer arm dynein in sea anemone, Anthopleura midori"Cell Motil. Cytoskel. 44. 202-208 (1999)

[Publications] Mohri, H.: "Tubulin 30 years later"Cell Struct. Funct.. 24. 265-267 (1999)

[Publications] Si, Y. and Okuno, M.: "Regulation of microtubule sliding by 36-kDa phosphoprotein in Hamster sperm flagella"Mol. Reprod. Dev.. 52. 328-334 (1999)

[Publications] Si, Y. and Okuno, M.: "Role of tyrosine phosphorylation of flagellar proteins in hamster sperm hyperactivation"Biol. Reprod.. 61. 240-246 (1999)

[Publications] 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)

[Publications] Ishijima, S. et al.: "Comparative studies on the spermatozoan motility of Japanese fishes"Bull. Ocean Res. Inst. Univ, Tokyo. 33. 139-152 (1998)

[Publications] Ishijima, S. et al.: "Movement of Turritella spermatozoa : direction of propagation and chirality of flagellar bends"Cell Motil. Cytoskel. 44. 85-95 (1999)

[Publications] 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)

[Publications] 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)

[Publications] Miyado, K. et al. (Okabe M.): "Requirement of CD9 on the egg plasma membrane for fertilization"Science. 287. 321-324 (2000)

[Publications] Hamada, Y. et al.: "Mutation in ankyrin repeats of the mouse notch2 gene induces early embryonic lethality"Development. 126. 7-10 (1997)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] Perry, A.C. et al. (Okabe M.): "Mammalian transgenesis by intracytoplasmic sperm injection"Science. 284. 1180-1183 (1999)

[Publications] 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)

[Publications] 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)

[Publications] Yanagida, K. et al. (Yanagimachi R.): "Successful fertilization and pregnancy following ICSI and electrical oocyte activation"Human Reprod. 14. 1307-1311 (1999)

[Publications] Kishikawa, H., Tateno, H. and Yanagimachi, R.: "Fertility of mouse spermatozoa from cadavers maintained at 4℃"J. Reprod. Feril. 116. 217-222 (1999)

[Publications] Cummins, J.M. et al. (Yanagimachi, R.): "Fate of genetically marked mitochondrial DNA from spermatocytes microinjected into mouse zygotes"Zygote. 7. 151-156 (1999)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] 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)

[Publications] Suarez, S.S.: "The Male Gemete : From basic science to clinical applications"Cache Riever Press. 10 (1999)

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)