1993 年度実績報告書

鞭毛・繊毛運動を制御する分子調節機構の解明

研究課題

研究課題/領域番号	04304006
研究機関	東京大学
研究代表者	森沢正昭東京大学, 理学部, 教授 (40013594)
研究分担者	奥野誠東京大学, 教養学部, 助教授 (40143325) 豊島陽子東京大学, 教養学部, 助教授 (40158043) 神谷律東京大学, 理学部, 教授 (10124314) 高橋景一国際基督教大学, 教養学部, 教授 (40011481) 毛利秀雄放送大学, 教養学部, 教授 (70012268)
キーワード	鞭毛・繊毛運動調節 / 微小管滑り / ダイニン / 再構成運動系 / モータードメイン / 遺伝子解析 / プロテアソーム / タンパク質リン酸化
研究概要	高橋は断片化したウニ精子鞭毛軸糸にエラスターゼを作用させ、ATP中で滑り運動を解析し、滑り速度のCa^<2+>依存性を示した。真行寺は除膜ウニ精子で、振動数を変えて鞭毛微小管滑り速度を解析し、滑り速度の振動数に伴う変化は外腕を抽出しても起きることを示した。豊島はキネシン頭部の遺伝子を用い特定部位を修飾し、頭部のみで微小管を滑らせる能力があることを明らかにした。小川はダイニンβ重鎖機能ドメインを、その相当領域をコードするcDNAをベクターにつなげ、バキュロウイルス発現系を用い量産する方法を確立した。神谷はクラミドモナスでダイニン外腕β鎖が短い変異株、内腕重鎖一部欠失株を単離、解析し、外腕の機能にβ重鎖が重要で、内腕重鎖の機能が重複していることを示した。毛利は系統上重要な腔腸動物イソギンチャクで精子鞭毛外腕ダイニンを生化学的・超微形態学的に検討し、2頭構造であることを示した。中村はキモトリプシン感受性の著しく異なるαチューブリン由来のチューブリンアイソフォームを見い出した。森沢は海産魚精子運動開始に細胞内イオン強度の増加が必要であることを示し、更にニシン精子活性化物質のcDNAクローニングを完成した。稲葉はシロサケ精子で、プロテアソームが鞭毛軸糸と細胞膜を連結している構造に局在していることを免疫電顕法により明らかにした。奥野は哺乳類精子で、繊維鞘の滑りが微小管#5-6のそれに伴って起きることから#5-6のダイニンの活性が高いことを示した。中岡は繊毛波の形成をcGMPが調節していることを示した。野口はゾウリムシ繊毛の有効打の方向制御機構に、29,65kDaタンパクのcAMP依存的リン酸化が重要であることを示した。馬場は精子鞭毛断片の周波数測定で、高Ca^<2+>でオシレーター2が特異的に阻害されることを示した。

研究成果

(52件)

すべてその他

すべて文献書誌 (52件)

[文献書誌] Morisawa,M.: "Regulatory mechanisms of sperm motility." In Biology of the germ line.In animals and man.(H.Mohri,M.Takahashi,and C.Tachi,eds.). 81-93 (1993)
[文献書誌] Morisawa,S.: "Roles of pH and cyclic adenosine monophosphate in the aquisition of potential for sperm motilityduring migration from the river in chum salmon." Mol.Rep.Dev.34. 420-460 (1993)
[文献書誌] Oda,S.: "Rises of intracelular Ca^<2+> and pH mediate the initiation of sperm motility by hyperosmolality in marine teleosts." Cell Motil.Cytoskeleton.25. 497-506 (1993)
[文献書誌] Yoshida.M.: "Sperm chemotaxix during the process of fertilization in the ascidians Ciona savygni and Ciona intestinalis." Dev.Biol.157. 497-506 (1993)
[文献書誌] Yamada,A.: "Sudden increase in speed of an filament moving on myosin cross-bridges of“mismatched"polarity observed when its leading end begins to interact with cross-bgidges of“matched"polarity." J.Biochem.111. 676-680 (1992)
[文献書誌] Murakami,A.: "Analysis of gravitational responce of Paramecium by free-all experiments." Proc.of the 9th Space Utilization Symp.102-104 (1992)
[文献書誌] Murakami,A.: "Analysis of gravitation and response to microgravity in Paramecium." Proc.of the 10th Space Utilization Symp.86-87 (1993)
[文献書誌] 高橋景一: "鞭毛運動" Annual Review細胞生物学1993. 165-178 (1993)
[文献書誌] Eshel,D.: "The phase of sperm flagellar beating is not conserved over a brief imposed interruption." Exp.Cell Res.202. 552-555 (1992)
[文献書誌] Ogawa,K.: "Primary structure and function of a dynein motor molecule." Zool.Sci.9. 265-274 (1992)
[文献書誌] 小川和男: "鞭毛モーターダイニンの分子構造" Cell Science. 9. 33-41 (1993)
[文献書誌] Ogawa,K.: "cDNA cloning for mouse caltractin" Biochim.Biophys.Acta.1216. 126-128 (1993)
[文献書誌] Kamimura,S.: "High-frequency vibration in flagellar axonemes with amplitudes reflecting the size of tubulin." J.Cell Biol.116. 1443-1454 (1992)
[文献書誌] Kamiya,R.: "Molecular mechanism of ciliary and flagellar movement" Advances in Comparative and Enviromental Physiology. 12. 206-226 (1992)
[文献書誌] Takada,S.: "Three-headed outer arm dynein from Chlamydomonas that can functionally combine with outer arm-missing axonemes." J.Biochem.111. 758-762 (1992)
[文献書誌] Kagami,O.: "Translocation and lotation of microtubules caused by multiple species of Chlamydomonas inner-arm dynein." J.Cell Biol.103. 653-664 (1992)
[文献書誌] Sakakibara,H.: "A Chlamydomonas outer arm dynein mutant with a truncated beta heavy chain." J.Cell Biol.122. 653-662 (1993)
[文献書誌] Kato,T.: "Isolation of two species of Chlamydomonas reinhardtii flagellar mutants,ida5 and ida6,that lack a newly identified heavy chain of the inner dynein arm" Cell Structure and Function. (印刷中). (1994)
[文献書誌] Johara,M.: "Charge-reversion mutagenesis of Dictyostelium actin to map the surface recognized by myosin during ATP-driven sliding motion" Proc.Natl.Acad.Sci.USA.90. 2127-2131 (1993)
[文献書誌] Itakura,S.: "Force-generating domain of myosin motor." Biochem.Biophys.Res.Commun.196. 1504-1510 (1993)
[文献書誌] 豊島陽子: "in vitro運動系" Annual Review細胞生物学1993. 3. 145-155 (1993)
[文献書誌] Iwamoto,T.: "Purification and characterization of a sperm motility-dynein ATPase inhibitor from boar seminal plasma." Mol.Reprod.Deveop.31. 55-62 (1992)
[文献書誌] Wada,S.: "Dynein of sperm flagella of oyster belonging to Protostomia also has a two headed structure" Biol.Cell. 76. 311-317 (1992)
[文献書誌] Mohri,H.: "Role of tubulin and dynein in spermatozoan motility" Mol.Reprod.Develop.36. 221-223 (1993)
[文献書誌] 高谷知恵子: "電気泳動法を応用したタンパク質の分離精製" 比較生理生化学. 9. 156-159 (1992)
[文献書誌] Wazawa,T.-I.: "Hydrogen peroxide plays a key role in the oxidation reaction of myoglobin by molecular oxygen." Biophys.J.63. 544-555 (1992)
[文献書誌] Jin,Z.X.: "Isolation and properties of a protein complex that cantains flagellar movement initiating-phosphoprotein from testis of chum salmon." J.Biochem.(印刷中). (1994)
[文献書誌] Jin,Z.X.: "Monoclonal antibodies against the protein complex that contains the flagellar movement initiating-phosphoprotein of Onchorhynchus keta." J.Biochem.(印刷中). (1994)
[文献書誌] 森沢正昭: "特集を終えるにあたって" Cell Science. 9. 1993 (1992)
[文献書誌] Inaba,K.: "Two high molecular mass proteases from sea urchin sperm" Biochem.Biophys.Res.Commun.182. 667-674 (1992)
[文献書誌] Inaba,K.: "Chymotrypsin-like protease activity associated with demembranated chum salmon sperm." Biol.Cell. 76. 329-333 (1992)
[文献書誌] 稲葉一男: "精子のプロテアソーム" Cell Science. 8. 750-758 (1992)
[文献書誌] Inaba,K.: "Interactions among the heterogenous components of dynein" In Comparative Spermatology 20 Years After(Baccetti,B.,ed.)Raven Press,New York.76. 385-386 (1992)
[文献書誌] Inaba,K.: "Purification of fproteasomes from salmonid fish sperm and their localization along sperm flagella." J.Cell Sci.104. 907-915 (1993)
[文献書誌] Mohri,H.: "Conformational changes of dynein molecule" In Comparative Spermatology 20 Years After(Baccetti,B.,ed.)Raven Press,New York.76. 413-416 (1992)
[文献書誌] Ishijima,S.: "Separation of X- and Y-chromosome-bearing sperm by free-flow electrophoresis:Evaluation of separation using PCR." Zool.Sci.9. 601-606 (1992)
[文献書誌] Ishijima,S.: "Identification of X- and Y-chromosome-bearing sperm separated by free-flow elecrophoresis using chromosome-specific polymerase chain reaction." Biomed.Res.13. 221-224 (1992)
[文献書誌] Ishida,K.: "Microflashing analysis of sperm motility." J.Reprod.Dev.39. 7-12 (1993)
[文献書誌] Si,Y.: "Multiple activation of mouse sperm motility." Mol.Reprod.Deveop.36. 89-95 (1993)
[文献書誌] Si,Y.: "The sliding of fibrous sheath through the axoneme proximally together with microtubule extrusion." Exp.Cell Res.208. 170-174 (1993)
[文献書誌] Nakaoka,Y.: "Hyperpolarization-activated inward current associated with the frequency increase in ciliary beating of Paramecium." J.Comp.Physiol.A. 170. 723-727 (1992)
[文献書誌] Nakaoka,Y.: "Cationic inhibition of Ca current and Ca-dependent ciliary responses in Paramecium." J.Comp.Physiol.A. (印刷中). (1994)
[文献書誌] Okamoto,K.: "Reconstitution of the metachronal waves in ciliated cortical sheet of Paramecium;I.Wave stabilities." J.Exp.Biol.(印刷中). (1994)
[文献書誌] Okamoto,K.: "Reconstitution of the metachronal waves in ciliated cortical sheet of Paramecium;II.Asymmetry of the ciliary movements." J.Exp.Biol.(印刷中). (1994)
[文献書誌] Noguchi,M.: "Effects of proteolytic digestion on the control mechanism of ciliary orientation in ciliated sheets from Paramecium." Zool.Sci.10. 267-273 (1993)
[文献書誌] Mogami,Y.: "Beating cilia of sea urchin embryos:Acritical comparison of the normal and reversed beating of cilia of isolated cells." J.Exp.Biol.175. 251-266 (1993)
[文献書誌] Mogami,Y.: "Regulation of ciliary movement in sea urchin embryos:Dopamine and 5-HT change the swimming behaviour." Comp.Biochem.Physiol.101C. 251-254 (1992)
[文献書誌] Ooya,M.: "Gravity-induced changes in propulsion of Paramecium:A possible role of gravireception in Protozoan behaviour." J.Exp.Biol.163. 153-167 (1992)
[文献書誌] Baba,S.A.: "High resolution analysis of flagellar bends." In Comparative Spermatology 20 Years After(Baccetti,B.,ed.)Raven Press,New York.75. 333-336 (1992)
[文献書誌] Toyoshima,Y.Y.: "Plenum Publishing Co.(eds.Sugi,H.and Pollack,G.H.)" How are myosin fragments bound to nitrocellulose film?In“Mechanism of myofilament sliding in muscle contraction"., 870(7) (1993)
[文献書誌] 毛利秀雄: "精子学" 東京大学出版会, 350 (1992)
[文献書誌] 神谷律: "細胞の運動" 培風館, 182 (1992)

1993 年度 実績報告書

鞭毛・繊毛運動を制御する分子調節機構の解明

研究代表者

森沢 正昭 東京大学, 理学部, 教授 (40013594)

研究成果

[文献書誌] Morisawa,M.: "Regulatory mechanisms of sperm motility." In Biology of the germ line.In animals and man.(H.Mohri,M.Takahashi,and C.Tachi,eds.). 81-93 (1993)

[文献書誌] Morisawa,S.: "Roles of pH and cyclic adenosine monophosphate in the aquisition of potential for sperm motilityduring migration from the river in chum salmon." Mol.Rep.Dev.34. 420-460 (1993)

[文献書誌] Oda,S.: "Rises of intracelular Ca^<2+> and pH mediate the initiation of sperm motility by hyperosmolality in marine teleosts." Cell Motil.Cytoskeleton.25. 497-506 (1993)

[文献書誌] Yoshida.M.: "Sperm chemotaxix during the process of fertilization in the ascidians Ciona savygni and Ciona intestinalis." Dev.Biol.157. 497-506 (1993)

[文献書誌] Yamada,A.: "Sudden increase in speed of an filament moving on myosin cross-bridges of“mismatched"polarity observed when its leading end begins to interact with cross-bgidges of“matched"polarity." J.Biochem.111. 676-680 (1992)

[文献書誌] Murakami,A.: "Analysis of gravitational responce of Paramecium by free-all experiments." Proc.of the 9th Space Utilization Symp.102-104 (1992)

[文献書誌] Murakami,A.: "Analysis of gravitation and response to microgravity in Paramecium." Proc.of the 10th Space Utilization Symp.86-87 (1993)

[文献書誌] 高橋景一: "鞭毛運動" Annual Review細胞生物学1993. 165-178 (1993)

[文献書誌] Eshel,D.: "The phase of sperm flagellar beating is not conserved over a brief imposed interruption." Exp.Cell Res.202. 552-555 (1992)

[文献書誌] Ogawa,K.: "Primary structure and function of a dynein motor molecule." Zool.Sci.9. 265-274 (1992)

[文献書誌] 小川和男: "鞭毛モーターダイニンの分子構造" Cell Science. 9. 33-41 (1993)

[文献書誌] Ogawa,K.: "cDNA cloning for mouse caltractin" Biochim.Biophys.Acta.1216. 126-128 (1993)

[文献書誌] Kamimura,S.: "High-frequency vibration in flagellar axonemes with amplitudes reflecting the size of tubulin." J.Cell Biol.116. 1443-1454 (1992)

[文献書誌] Kamiya,R.: "Molecular mechanism of ciliary and flagellar movement" Advances in Comparative and Enviromental Physiology. 12. 206-226 (1992)

[文献書誌] Takada,S.: "Three-headed outer arm dynein from Chlamydomonas that can functionally combine with outer arm-missing axonemes." J.Biochem.111. 758-762 (1992)

[文献書誌] Kagami,O.: "Translocation and lotation of microtubules caused by multiple species of Chlamydomonas inner-arm dynein." J.Cell Biol.103. 653-664 (1992)

[文献書誌] Sakakibara,H.: "A Chlamydomonas outer arm dynein mutant with a truncated beta heavy chain." J.Cell Biol.122. 653-662 (1993)

[文献書誌] Kato,T.: "Isolation of two species of Chlamydomonas reinhardtii flagellar mutants,ida5 and ida6,that lack a newly identified heavy chain of the inner dynein arm" Cell Structure and Function. (印刷中). (1994)

[文献書誌] Johara,M.: "Charge-reversion mutagenesis of Dictyostelium actin to map the surface recognized by myosin during ATP-driven sliding motion" Proc.Natl.Acad.Sci.USA.90. 2127-2131 (1993)

[文献書誌] Itakura,S.: "Force-generating domain of myosin motor." Biochem.Biophys.Res.Commun.196. 1504-1510 (1993)

[文献書誌] 豊島陽子: "in vitro運動系" Annual Review細胞生物学1993. 3. 145-155 (1993)

[文献書誌] Iwamoto,T.: "Purification and characterization of a sperm motility-dynein ATPase inhibitor from boar seminal plasma." Mol.Reprod.Deveop.31. 55-62 (1992)

[文献書誌] Wada,S.: "Dynein of sperm flagella of oyster belonging to Protostomia also has a two headed structure" Biol.Cell. 76. 311-317 (1992)

[文献書誌] Mohri,H.: "Role of tubulin and dynein in spermatozoan motility" Mol.Reprod.Develop.36. 221-223 (1993)

[文献書誌] 高谷知恵子: "電気泳動法を応用したタンパク質の分離精製" 比較生理生化学. 9. 156-159 (1992)

[文献書誌] Wazawa,T.-I.: "Hydrogen peroxide plays a key role in the oxidation reaction of myoglobin by molecular oxygen." Biophys.J.63. 544-555 (1992)

[文献書誌] Jin,Z.X.: "Isolation and properties of a protein complex that cantains flagellar movement initiating-phosphoprotein from testis of chum salmon." J.Biochem.(印刷中). (1994)

[文献書誌] Jin,Z.X.: "Monoclonal antibodies against the protein complex that contains the flagellar movement initiating-phosphoprotein of Onchorhynchus keta." J.Biochem.(印刷中). (1994)

[文献書誌] 森沢正昭: "特集を終えるにあたって" Cell Science. 9. 1993 (1992)

[文献書誌] Inaba,K.: "Two high molecular mass proteases from sea urchin sperm" Biochem.Biophys.Res.Commun.182. 667-674 (1992)

[文献書誌] Inaba,K.: "Chymotrypsin-like protease activity associated with demembranated chum salmon sperm." Biol.Cell. 76. 329-333 (1992)

[文献書誌] 稲葉一男: "精子のプロテアソーム" Cell Science. 8. 750-758 (1992)

[文献書誌] Inaba,K.: "Interactions among the heterogenous components of dynein" In Comparative Spermatology 20 Years After(Baccetti,B.,ed.)Raven Press,New York.76. 385-386 (1992)

[文献書誌] Inaba,K.: "Purification of fproteasomes from salmonid fish sperm and their localization along sperm flagella." J.Cell Sci.104. 907-915 (1993)

[文献書誌] Mohri,H.: "Conformational changes of dynein molecule" In Comparative Spermatology 20 Years After(Baccetti,B.,ed.)Raven Press,New York.76. 413-416 (1992)

[文献書誌] Ishijima,S.: "Separation of X- and Y-chromosome-bearing sperm by free-flow electrophoresis:Evaluation of separation using PCR." Zool.Sci.9. 601-606 (1992)

[文献書誌] Ishijima,S.: "Identification of X- and Y-chromosome-bearing sperm separated by free-flow elecrophoresis using chromosome-specific polymerase chain reaction." Biomed.Res.13. 221-224 (1992)

[文献書誌] Ishida,K.: "Microflashing analysis of sperm motility." J.Reprod.Dev.39. 7-12 (1993)

[文献書誌] Si,Y.: "Multiple activation of mouse sperm motility." Mol.Reprod.Deveop.36. 89-95 (1993)

[文献書誌] Si,Y.: "The sliding of fibrous sheath through the axoneme proximally together with microtubule extrusion." Exp.Cell Res.208. 170-174 (1993)

[文献書誌] Nakaoka,Y.: "Hyperpolarization-activated inward current associated with the frequency increase in ciliary beating of Paramecium." J.Comp.Physiol.A. 170. 723-727 (1992)

[文献書誌] Nakaoka,Y.: "Cationic inhibition of Ca current and Ca-dependent ciliary responses in Paramecium." J.Comp.Physiol.A. (印刷中). (1994)

[文献書誌] Okamoto,K.: "Reconstitution of the metachronal waves in ciliated cortical sheet of Paramecium;I.Wave stabilities." J.Exp.Biol.(印刷中). (1994)

[文献書誌] Okamoto,K.: "Reconstitution of the metachronal waves in ciliated cortical sheet of Paramecium;II.Asymmetry of the ciliary movements." J.Exp.Biol.(印刷中). (1994)

[文献書誌] Noguchi,M.: "Effects of proteolytic digestion on the control mechanism of ciliary orientation in ciliated sheets from Paramecium." Zool.Sci.10. 267-273 (1993)

[文献書誌] Mogami,Y.: "Beating cilia of sea urchin embryos:Acritical comparison of the normal and reversed beating of cilia of isolated cells." J.Exp.Biol.175. 251-266 (1993)

[文献書誌] Mogami,Y.: "Regulation of ciliary movement in sea urchin embryos:Dopamine and 5-HT change the swimming behaviour." Comp.Biochem.Physiol.101C. 251-254 (1992)

[文献書誌] Ooya,M.: "Gravity-induced changes in propulsion of Paramecium:A possible role of gravireception in Protozoan behaviour." J.Exp.Biol.163. 153-167 (1992)

[文献書誌] Baba,S.A.: "High resolution analysis of flagellar bends." In Comparative Spermatology 20 Years After(Baccetti,B.,ed.)Raven Press,New York.75. 333-336 (1992)

[文献書誌] Toyoshima,Y.Y.: "Plenum Publishing Co.(eds.Sugi,H.and Pollack,G.H.)" How are myosin fragments bound to nitrocellulose film?In“Mechanism of myofilament sliding in muscle contraction"., 870(7) (1993)

[文献書誌] 毛利秀雄: "精子学" 東京大学出版会, 350 (1992)

[文献書誌] 神谷律: "細胞の運動" 培風館, 182 (1992)

1993 年度実績報告書

森沢正昭東京大学, 理学部, 教授 (40013594)