1992 Fiscal Year Annual Research Report

神経細胞・細胞骨格の分子細胞生物学的研究

Research Project

Project/Area Number	04102005
Research Institution	The University of Tokyo
Principal Investigator	廣川信隆東京大学, 医学部, 教授 (20010085)
Co-Investigator(Kenkyū-buntansha)	吉武玲子東京大学, 医学部, 助手 (40230718) 中田隆夫東京大学, 医学部, 助手 (50218004) 金井克光東京大学, 医学部, 助手 (80214427) 岡部繁男東京大学, 医学部, 助手 (60204012) 依藤宏東京大学, 医学部, 講師 (00158544)
Keywords	MAP2 / MAP2C / Tau / 新しいモータ分子 / 神経細胞 / 細胞骨格 / 形態形成 / 物質輸送
Research Abstract	1〕形態形成(1)樹状突起に特異に発現されるMAP2(1828aa)と軸索に多量に発現されるMAP2C(467aa)とTau(432aa)の機能をバキュロウィルスSf9Cell系を用いて解析した。N末Projection domainの長さの差により樹状突起や軸索に特異な微小管領域が形成されることがわかった。(2)MAPの微小管のdynamicsへ与える影響を調べるため、MAP2CcDNAを形質導入した線維芽細胞へCaged fluorescein label tubulinを微量注入し紫外線照射によるphotoactivation法により微小管のターンオーバーを解析し、MAP2Cが微小管を安定化する事を明らかにした。 2〕物質輸送(1)神経軸索内の物質輸送の新しいモーター分子を分子生物学的手法を用いて探索し、従来同定されていたキネシンと脳ダイニン以外の5種類の新しいモーター分子を同定し、その内4種類のcDNAクローニングと一次構造の決定を行い。in situ hybridization法によりこの5類種がすべて一つの神経細胞に発現されている事を明らかにした。(2)神経軸索内のニューロフィラメントの動態と遅い軸索輸送の機構について、ニューロフィラメントL蛋白にfluorescein標識し、培養神経細胞に微量注入その後のレーザー光によりphotobleach recovery法で解析した。その結果ニューロフィラメントは軸索内でターンオーバーするdynamicな構造で、その輸送はpalymerの形でなくsmalloligomerとして動いている事が強く示唆された。 3〕シナプス領域での伝達物質の放出及び小胞膜のrecyclingの機構シナプス小胞のrecyclingの過程で重要な機能が予想されるdynamin(100KD)α大量精製法を確立し、これが微小管により活性化されるGTPase活性を有し、免疫細胞化学により順行性に運ばれる膜小器官に結合してシナプス結末に運ばれる事を示した。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] Takeuchi,M.: "The 72KD microtubule associated protein form porcine brain." Journal of Neurochemistry. 58. 1510-1516 (1992)
[Publications] Okabe,S.: "Differential behavior of photoactivated microtubules in growing axon of mouse and frog neurons." Journal of Cell Biology. 117. 105-120 (1992)
[Publications] Hirokawa,N.: "Microtubules on the move ?" Current Biology. 2. 193-195 (1992)
[Publications] Nakata,T.: "Ordanization of cortical cytoskeleton of cultured chromaff in cells and involvement in secretion as revealed by quick-freeze,deep-etchig,and double-label immunoelectrom microscopy." Journal of Neuroscience. 12. 2186-2197 (1992)
[Publications] Oshima,T.: "Immunocytochemical localization of 205 kDa microtubule-associated protein (205kDa MAP) in the guinea pig organ of corti." Brain Research. 590. 53-65 (1992)
[Publications] Maeda,K.: "Dynamin:its enzymatic activity and interaction with microtubules." Molecular Biology of the Cell. 3. 1181-1194 (1992)
[Publications] Takemura,R.: "Reorganization of brain spectrin (fodrin) during differentiation of PC12 cells." Neuroscience. 52. 381-391 (1993)
[Publications] Umeyama,T.: "Dynamics of microtubules bundled by microtubule associated protein 2 C (MAP2)." Journal of Cell Biology. 120. 451-465 (1993)
[Publications] Okabe,S.: "Do photobleached fluorescent microtubules move ? Re-evaluation of fluorescence laser photobleaching both in vitro and in growing Xenopus axon." Journal of Cell Biology. (1993)
[Publications] Noda,Y.: "Localization association with membraneous organelles transported of dynamin:Widespread distribution in neurons and anterogradely." Neuroscience. (1993)
[Publications] Okabe,S.: "Dynamics of the neuronal intermediate filaments." Journal of Cell. (1993)
[Publications] Nakata,T.: "T-dynamin,a novel member of dynamin-family protein is expressed specific to testis." Journal of Cell Science. (1993)
[Publications] Kanai,Y.: "Microtubule bundling by tau proteins in vivo:analysis of the functional domains." EMBO Journal. 11. 3953-3961 (1992)
[Publications] Aizawa,H.: "Kinesin family in murine central nervous system." Journal of Cell Biology. 119. 1287-1296 (1992)
[Publications] Tanaka,Y.: "Chronological expression of microtubule-associated proteins (MAPs) in EC cell p 19 after neuronal inductin by retinoic acid." Brain Research. 596. 269-278 (1992)
[Publications] Sato-Yoshitake: "Phosphorylation of kinesin reguiates its binding to synaptic vesicles." Journal of Biological Chemistry. 267. 23930-23936 (1992)
[Publications] Chen,J.: "Projection domains of MAP2 and tau determine spacings between microtubules in dendrites and axons." Nature. 360. 674-677 (1992)
[Publications] Takemura,R.: "Increased microtubule stability and alpha tubulin acetylation in cells transfected with microtubule-associated proteins MAP1B,MAP2 or tau." Journal of Cell Science. 103. 953-964 (1992)

1992 Fiscal Year Annual Research Report

神経細胞・細胞骨格の分子細胞生物学的研究

Principal Investigator

廣川 信隆 東京大学, 医学部, 教授 (20010085)

Research Products

[Publications] Takeuchi,M.: "The 72KD microtubule associated protein form porcine brain." Journal of Neurochemistry. 58. 1510-1516 (1992)

[Publications] Okabe,S.: "Differential behavior of photoactivated microtubules in growing axon of mouse and frog neurons." Journal of Cell Biology. 117. 105-120 (1992)

[Publications] Hirokawa,N.: "Microtubules on the move ?" Current Biology. 2. 193-195 (1992)

[Publications] Nakata,T.: "Ordanization of cortical cytoskeleton of cultured chromaff in cells and involvement in secretion as revealed by quick-freeze,deep-etchig,and double-label immunoelectrom microscopy." Journal of Neuroscience. 12. 2186-2197 (1992)

[Publications] Oshima,T.: "Immunocytochemical localization of 205 kDa microtubule-associated protein (205kDa MAP) in the guinea pig organ of corti." Brain Research. 590. 53-65 (1992)

[Publications] Maeda,K.: "Dynamin:its enzymatic activity and interaction with microtubules." Molecular Biology of the Cell. 3. 1181-1194 (1992)

[Publications] Takemura,R.: "Reorganization of brain spectrin (fodrin) during differentiation of PC12 cells." Neuroscience. 52. 381-391 (1993)

[Publications] Umeyama,T.: "Dynamics of microtubules bundled by microtubule associated protein 2 C (MAP2)." Journal of Cell Biology. 120. 451-465 (1993)

[Publications] Okabe,S.: "Do photobleached fluorescent microtubules move ? Re-evaluation of fluorescence laser photobleaching both in vitro and in growing Xenopus axon." Journal of Cell Biology. (1993)

[Publications] Noda,Y.: "Localization association with membraneous organelles transported of dynamin:Widespread distribution in neurons and anterogradely." Neuroscience. (1993)

[Publications] Okabe,S.: "Dynamics of the neuronal intermediate filaments." Journal of Cell. (1993)

[Publications] Nakata,T.: "T-dynamin,a novel member of dynamin-family protein is expressed specific to testis." Journal of Cell Science. (1993)

[Publications] Kanai,Y.: "Microtubule bundling by tau proteins in vivo:analysis of the functional domains." EMBO Journal. 11. 3953-3961 (1992)

[Publications] Aizawa,H.: "Kinesin family in murine central nervous system." Journal of Cell Biology. 119. 1287-1296 (1992)

[Publications] Tanaka,Y.: "Chronological expression of microtubule-associated proteins (MAPs) in EC cell p 19 after neuronal inductin by retinoic acid." Brain Research. 596. 269-278 (1992)

[Publications] Sato-Yoshitake: "Phosphorylation of kinesin reguiates its binding to synaptic vesicles." Journal of Biological Chemistry. 267. 23930-23936 (1992)

[Publications] Chen,J.: "Projection domains of MAP2 and tau determine spacings between microtubules in dendrites and axons." Nature. 360. 674-677 (1992)

[Publications] Takemura,R.: "Increased microtubule stability and alpha tubulin acetylation in cells transfected with microtubule-associated proteins MAP1B,MAP2 or tau." Journal of Cell Science. 103. 953-964 (1992)

廣川信隆東京大学, 医学部, 教授 (20010085)