1989 Fiscal Year Annual Research Report

細胞骨格の分子構築と機能〜細胞生物学及び分子生物学的研究

Research Project

Project/Area Number	62065007
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)	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) 久永真市東京大学, 医学部, 助手 (20181092) 依藤宏東京大学, 医学部, 講師 (00158544)
Keywords	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
Research Abstract	1.神経細胞の主要な微小管関連蛋白(MAPs)の1つである(MAP1B)をモノクロナール抗体吸着カラムを用いて単離しその単分子構造、細胞内局在を明らかとした。又リン酸化形MAP1Bを認識するモノクローン抗体を作製しリン酸化形は軸索に多いが、脱リン酸化型は樹状突起に多い事及び特に発生初期、軸索の形成に伴いリン酸化MAP1Bが発現される事を明らかとした。 2.シナプスにおける小胞分泌のモデルとして初代培養副脊髄質細胞を用い、Ca^<++>流入に伴うCa^<++>感受性蛋白の関与を検討した。リポコルチン族のカルパクチン1の分子構造及びその抗体を用いた免疫細胞化学によりカルパクチン1が形質膜内側に局在しCa^<++>流入に伴い分子形態を変化させ分泌顆粒膜と形質膜を癒合させ分泌をうながす事を明らかとした。 3.ニューロフィラメント内でL蛋白単分子がどの様に配列して一本のフィラメントを形成するのか、又長さの伸長にはNa^+イオンが単分子の側方の結合にはMg^<++>イオンが重要である事などassemblyの機構を明らかとした。又、cAMP依存性カイネースやCカイネースによるL蛋白のN末部位のリン酸化によりニューロフィラメントが8量体にばらけ、又8量体から一本のフィラメントの重合が阻害される事が明らかとなった。 4.初代培養神経細胞へ蛍光標識チュブリン及びアクチンを微量注入しその後、レーザー光による蛍光消褪を行いその後の回復を観察する事によってチュブリン及びアクチンの遅い輸送が従来考えられていた様に重合体が動くのではなくダイマーないしオリゴマーの形で運ばれて軸索局所で新しい線維に組み込まれる事を明らかとした。

Research Products

(24 results)

All Other

All Publications (24 results)

[Publications] Hirokawa,N.: "The arrangement of actin filaments in the postsynaptic cytoplasm revealed with the quick freeze deep etch electron microscopy." Neuroscience Research.6. 269-275 (1989)
[Publications] Hirokawa,N.: "The cytoskeletal architecture of the presynaptic terminal and molecular structure of synapsin 1." Journal of Cell Biology. 108. 111-126 (1989)
[Publications] Okabe,S.: "Axonal transport." In“Current Opininon in Cell Biology,Vol.1,no.1 editors M.Kirschner,and K.Weber,Gower Academic Journal. 91-97 (1989)
[Publications] Yorifuji,H.: "Localization of 4.1 related proteins in cerebellar neurons." European Journal of Cell Biology. 48. 104-115 (1989)
[Publications] Okabe,S.: "Immunocytochemical localization of microtubule-associated proteins 1A and 2 in the rat retina." Brain Research. 483. 335-346 (1989)
[Publications] Hirokawa,N.: "Submolecular domains of bovine brain kinesin identified by electron microscopy and monoclonal antibody decoration." Cell. 56. 867-878 (1989)
[Publications] Sato,R.: "Molecular structure and localization of microtubuleーassociated protein 1B and its phosphorylation-dependent expression in the developing neurons." Neuron. 3. 229-238 (1989)
[Publications] Maekawa,S.: "Purification and characterization of a Ca^<2+>-dependent actin filament severing protein from bovine adrenal medulla." Journal of Biological Chemistry. 264. 7458-7465 (1989)
[Publications] Okabe,S.: "Incorporation and turnover of biotin-labeled actin microinjected into fibroblastic cells:An immunoelectron microscopic study." Journal of Cell Biology. 109. 1581-1595 (1989)
[Publications] Shiomura,Y.: "A novel heat-stable 205KD microtubule-associated protein is involved in the neural development." Brain Research. 502. 356-364 (1989)
[Publications] Nakata,T.: "Conformational change and localization of calpactin 1 complex involved in exocytosis as revealed by quickーfreeze,deep-etch technique and immunocytochemistry." Journal of Cell Biology. 110. 13-25 (1990)
[Publications] Okabe,S.: "Turnover of fluorescently labeled tubulin and actin in the axon." Nature. 343. 479-482 (1990)
[Publications] Hirokawa,N.: "Cytoskeletal architecture of the mitotic spindle." “Cell Movement vol.11 Kinesin,Dynein and Microtubule Dynamics."R.McIntosh and F.Warner editors Alan R.Liss Inc.New York,. 2. 383-402 (1989)
[Publications] Hisanaga,S.: "The effects of dephosphorylation on the structure of the projections of neurofilament." Journal of Neuroscience. 9. 959-966 (1989)
[Publications] Hisanaga,S.: "The molecular structure of adrenal medulla kinesin." Cell Motility and Cytoskeleton.12. 264-272 (1989)
[Publications] Okabe,S.: "Rapid turnover of microtubule-associated protein 2 in the axon." Proc.Natl Acad Sci.USA. 86. 4127-4131 (1989)
[Publications] Yorifuji,H.: "Cytoskeletal architecture of neuromuscular junction-Localization of vinculin." Journal of Electron Microscopic Technique. 12. 160-171 (1989)
[Publications] Kanai,Y.: "Expression of multiple tau isoforms and microtubule bundle formation in fibroblasts transfected with a single tau cDNA." Journal of Cell Biology.109. 1173-1184 (1989)
[Publications] Hisanaga,S.: "Molecular architecture of neurofilament.I.Subunit arrangement of neurofilament-L protein in the intermediate sized filament." Journal of Molecular Biology. (1990)
[Publications] Hisanaga,S.: "Molecular architecture of neurofilament.II.In vitro reassembly process of neurofilament-L protein." Journal of Molecular Biology. (1990)
[Publications] Hisanaga,S.: "Effects of phosphorylation of the neurofilament L protein on filamentous structures." Cell Regulation. (1990)
[Publications] Takemura,R.: "Developmental expression of tau mRNAs in rat brain." (1990)
[Publications] Harada,A.: "Developmental changes of synapsin 1 expression in the rat cerebellar neurons." (1990)
[Publications] Hirokawa,N.: "Brain dynein(MAP1C)localizes on both anterogradely and retrogradely transported membranous organelles." (1990)

1989 Fiscal Year Annual Research Report

細胞骨格の分子構築と機能〜細胞生物学及び分子生物学的研究

Principal Investigator

廣川 信隆 東京大学, 医学部, 教授 (20010085)

Research Products

[Publications] Hirokawa,N.: "The arrangement of actin filaments in the postsynaptic cytoplasm revealed with the quick freeze deep etch electron microscopy." Neuroscience Research.6. 269-275 (1989)

[Publications] Hirokawa,N.: "The cytoskeletal architecture of the presynaptic terminal and molecular structure of synapsin 1." Journal of Cell Biology. 108. 111-126 (1989)

[Publications] Okabe,S.: "Axonal transport." In“Current Opininon in Cell Biology,Vol.1,no.1 editors M.Kirschner,and K.Weber,Gower Academic Journal. 91-97 (1989)

[Publications] Yorifuji,H.: "Localization of 4.1 related proteins in cerebellar neurons." European Journal of Cell Biology. 48. 104-115 (1989)

[Publications] Okabe,S.: "Immunocytochemical localization of microtubule-associated proteins 1A and 2 in the rat retina." Brain Research. 483. 335-346 (1989)

[Publications] Hirokawa,N.: "Submolecular domains of bovine brain kinesin identified by electron microscopy and monoclonal antibody decoration." Cell. 56. 867-878 (1989)

[Publications] Sato,R.: "Molecular structure and localization of microtubuleーassociated protein 1B and its phosphorylation-dependent expression in the developing neurons." Neuron. 3. 229-238 (1989)

[Publications] Maekawa,S.: "Purification and characterization of a Ca^<2+>-dependent actin filament severing protein from bovine adrenal medulla." Journal of Biological Chemistry. 264. 7458-7465 (1989)

[Publications] Okabe,S.: "Incorporation and turnover of biotin-labeled actin microinjected into fibroblastic cells:An immunoelectron microscopic study." Journal of Cell Biology. 109. 1581-1595 (1989)

[Publications] Shiomura,Y.: "A novel heat-stable 205KD microtubule-associated protein is involved in the neural development." Brain Research. 502. 356-364 (1989)

[Publications] Nakata,T.: "Conformational change and localization of calpactin 1 complex involved in exocytosis as revealed by quickーfreeze,deep-etch technique and immunocytochemistry." Journal of Cell Biology. 110. 13-25 (1990)

[Publications] Okabe,S.: "Turnover of fluorescently labeled tubulin and actin in the axon." Nature. 343. 479-482 (1990)

[Publications] Hirokawa,N.: "Cytoskeletal architecture of the mitotic spindle." “Cell Movement vol.11 Kinesin,Dynein and Microtubule Dynamics."R.McIntosh and F.Warner editors Alan R.Liss Inc.New York,. 2. 383-402 (1989)

[Publications] Hisanaga,S.: "The effects of dephosphorylation on the structure of the projections of neurofilament." Journal of Neuroscience. 9. 959-966 (1989)

[Publications] Hisanaga,S.: "The molecular structure of adrenal medulla kinesin." Cell Motility and Cytoskeleton.12. 264-272 (1989)

[Publications] Okabe,S.: "Rapid turnover of microtubule-associated protein 2 in the axon." Proc.Natl Acad Sci.USA. 86. 4127-4131 (1989)

[Publications] Yorifuji,H.: "Cytoskeletal architecture of neuromuscular junction-Localization of vinculin." Journal of Electron Microscopic Technique. 12. 160-171 (1989)

[Publications] Kanai,Y.: "Expression of multiple tau isoforms and microtubule bundle formation in fibroblasts transfected with a single tau cDNA." Journal of Cell Biology.109. 1173-1184 (1989)

[Publications] Hisanaga,S.: "Molecular architecture of neurofilament.I.Subunit arrangement of neurofilament-L protein in the intermediate sized filament." Journal of Molecular Biology. (1990)

[Publications] Hisanaga,S.: "Molecular architecture of neurofilament.II.In vitro reassembly process of neurofilament-L protein." Journal of Molecular Biology. (1990)

[Publications] Hisanaga,S.: "Effects of phosphorylation of the neurofilament L protein on filamentous structures." Cell Regulation. (1990)

[Publications] Takemura,R.: "Developmental expression of tau mRNAs in rat brain." (1990)

[Publications] Harada,A.: "Developmental changes of synapsin 1 expression in the rat cerebellar neurons." (1990)

[Publications] Hirokawa,N.: "Brain dynein(MAP1C)localizes on both anterogradely and retrogradely transported membranous organelles." (1990)

廣川信隆東京大学, 医学部, 教授 (20010085)