| Project/Area Number |
62065007
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| Research Category |
Grant-in-Aid for Specially Promoted Research
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| Allocation Type | Single-year Grants |
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| Research Institution | University of Tokyo |
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Principal Investigator |
HIROKAWA Nobutaka Sch. of Med, Univ. of Tokyo, Professor, 医学部(医), 教授 (20010085)
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| Co-Investigator(Kenkyū-buntansha) |
HARADA Reiko Sch. of Med, Univ. of Tokyo, Assistant, 医学部(医), 助手 (40230718)
NAKATA Takao Sch. of Med, Univ. of Tokyo, Assistant, 医学部(医), 助手 (50218004)
KANAI Yoshimitsu Sch. of Med, Univ. of Tokyo, Assistant, 医学部(医), 助手 (80214427)
OKABE Shigeo Sch. of Med, Univ. of Tokyo, Assistant, 医学部(医), 助手 (60204012)
YORIFUJI Hiroshi Sch. of Med, Univ. of Tokyo, Lecturer, 医学部(医), 講師 (00158544)
久永 真市 東京大学, 医学部, 助手 (20181092)
竹村 玲子 沖中記念成人病研究所, 研究員 (50171674)
塩村 洋子 東京大学, 医学部解剖学教室, 助手 (00136962)
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| Project Period (FY) |
1987 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥243,000,000 (Direct Cost: ¥243,000,000)
Fiscal Year 1991: ¥30,000,000 (Direct Cost: ¥30,000,000)
Fiscal Year 1990: ¥30,000,000 (Direct Cost: ¥30,000,000)
Fiscal Year 1989: ¥33,000,000 (Direct Cost: ¥33,000,000)
Fiscal Year 1988: ¥60,000,000 (Direct Cost: ¥60,000,000)
Fiscal Year 1987: ¥90,000,000 (Direct Cost: ¥90,000,000)
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| Keywords | Neuron / Cytoskeleton / Microtubule / MAPs / Morphogenesis / Axonal transpsort / Kinesin / Synaptic Transmission / タウ蛋白 / 微小管関連蛋白MAP1B / 副腎髄質細胞 / 伝達物質放出 / カルパクチン1 / ニューロフィラメントの重合メカニズム / ニューロフィラメントリン酸化 / 蛍光消褪法 / 軸索輸送のメカニズム / 分子構築と動態 / ニューロフィラメントアクチン / シナプス / ニューロフィラメント |
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| Research Abstract |
(1) We purified the microtubule associated protein 2C, which specifically expressed in juvenile brains, and characterized them biochemically and ultrastructurally. The MAP2C CDNA was cloned from rat brain CDNA library and was sequenced. To analyze functions of MAP2 in vivo, we carried out transfection experiments. A. fibroblast cell line which was transfected with the CDNA expressed MAP2C and formed thick bundles of 'microtubules. Ineterestingly intermediate filaments also changed their organization and colocalized with microtubule bundles.
(2) To elucidate the mechamism which control bidirectional transport of membranous organelle we examined the effect of phosphorylation of kinesin on their binding to the membranes ; We found phosphorylation of kinesin by A-kinase considerably decreased the binding. Thus A-kinase probably play an important role on the dissociation of kinesin from the membranous, organelles in the. axon.
(3) To study the dynamics of tubulin in axons we microinjected caged fluorescein labeled tubulin into cultured neurons and subsequently analysed its turnover by UV photoactivation method. The results suggested that most of the polymers in the axon are stationary and the free oligomers could be the form to be transported.
(4) We established a method to purify 10OKD microtubule Associated idrotein (dynamin) from mammalian brains. Dynamin revealed a potent GTPase activity activated by microtubules. CDNA of dynamin was cloned and sequenced. Dynaim was proved to be a new GTP binding protein which specifically expressed in mature nerve cells.
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