1991 Fiscal Year Annual Research Report

細胞骨格の分子講築と機能〜細胞生物学及び分子生物学的研究

Research Project

Project/Area Number	62065007
Research Institution	University of Tokyo
Principal Investigator	廣川信隆東京大学, 医学部(医), 教授 (20010085)
Co-Investigator(Kenkyū-buntansha)	吉武玲子東京大学, 医学部(医), 助手 (40230718) 中田隆夫東京大学, 医学部(医), 助手 (50218004) 金井克光東京大学, 医学部(医), 助手 (80214427) 岡部繁男東京大学, 医学部(医), 助手 (60204012) 依藤宏東京大学, 医学部(医), 講師 (00158544)
Keywords	神経細胞 / 細胞骨格 / 微小管 / 微小管関連蛋白 / 形態形成 / 軸索輸送 / キネシン
Research Abstract	(1)神経細胞の幼若期に特に多量発現され、主要な微小管関連蛋白(MAP_s)の一つであるMAP_2Cの精製を確立し生化学的性状を解析し、 _cDNAのクロ-ニングを行った。一次構造を決め _cDNAを線維芽細胞に形質導入すると線維芽細胞内に微小管の束が形成され突起の中に伸長した。又この細胞で中間径線維も微小管と平行に配列する事が分かりMAP_2が微小管の重合促進と束化だけでなく、中間径線維を配行させる機能をもつ事が示された。(2)軸索内の順行性及び逆行性の膜小器官輸送の制御機構として順行性膜輸送のモ-タ-であるキネシンがAキナ-ゼでリン酸化を受けると膜小器官からはずれる事を示し、これが終末部でキネシンが順行性膜小器官から解離するため、重要な役割りをはたしている事が示唆された。(3)細胞骨格蛋白の遅い輸送と関連して、神経細胞内のチュブリンの動態を、Caged fluorescein標識チュブリンを初代培養神経細胞への微量注入その後の紫外線照射による蛍光活性化法によって解析し、哺乳類神経細胞ではチュブリン等の細胞骨格蛋白は、重合体の滑し運動でなくフリ-のオリゴマ-の形で輸送される事を示した。しかし両生類神経細胞等のごく一部の非常に突起伸長速度の速い細胞では、重合体が軸索膜と一体として成長端により引っ張られ受動的に動く現象もみられた。(4)哺乳類脳から、ヌクレオチド依存性に、微小管との結合が変化する100KDの蛋白ダイナミンを生化学的に大量精製する方法を確立した。その結果ダイナミンは、微小管により活性化されるGTPaseである事が分かった。さらにダイナミンCDNAはクロ-ニングを行いその一次構造を決定した。その結果ダイナミンは新しいGTP結合蛋白である事が判明しまた神経細胞特に成熟した神経細胞に多く発現される蛋白である事が分かった。

Research Products

(17 results)

All Other

All Publications (17 results)

[Publications] Hirokawa,N.: "Molecular architecture and dynamics of the neuronal cytoskeleton.in Neuronal cytoskeleton." Wily Liss,Inc.,New York. 5-74 (1991)
[Publications] Okabe,s.: "Actin dynamics in growth cones." J.of Neuroscience. 11. 1918-1929 (1991)
[Publications] Hirokawa,N.: "Kinesin associates with anterogradely transported membraous organelles." J.of Cell biology. 114. 259-302 (1991)
[Publications] Nakata,T.: "Predominant and developmentally regulated expression of dynamin in neurons." Neuron. 7. 461-469 (1991)
[Publications] Takemura,R.: "Developmental expression of tau mRNAs in rat brain." Neuroscience. 44. 393-407 (1991)
[Publications] Takeuchi,M.: "The 72KD microtubule associated protein from porcine brain." J.of Neurochemistry. (1992)
[Publications] Nakata,T.: "Organization of cortical cytoskeleton of cultured chromaffin cells and involvement in secretion as revealed by quick-freeze,deep-etching,and double-label immunoelectron microscopy." J.of Neuroscience. (1992)
[Publications] Okabe,S.: "Differential behavior of photoactivated microtubules in qrowing axon of mouse and frog neurons." J.of Cell Biology,. (1992)
[Publications] Kanai,Y.: "Microtubule bundling by tau proteins in vivo:analysis of the functional domains."
[Publications] Sato-Yoshitake,R.: "Phosphorylation of kinesin regulates its binding to synaptic vesicles."
[Publications] Takemura,R.: "Reorganization of brain spectrin (fodrin) during differentiation of PC12 cells."
[Publications] Okada,Y.: "Functional differentiation of axonal microtubules;Tyrosinated alpha-tubulin rich microtubules for transport,and detyrosinated alpha-tubulin rich microtubules for architecture."
[Publications] Noda,Y.: "Localization of dynamin:Widespread distribution in neurons and association with membraneous organelles transported anterogradely."
[Publications] Nakata,T.: "T-dynamin,a novel member of dynamin-family protein is expressed specific to testis."
[Publications] Maeda,K.: "Dynamin:its enzymatic activity and interaction with microtubules."
[Publications] Oshima,T.: "Immunocytochemical localization of 205 kDa microtubule-associated protein (205kDa MAP) in the guinea pig organ of corti."
[Publications] Hirokawa,N.: "Selective stabilization of microinjected tau in the axon and microtubule associated proteins 2 (MAP2) in dendrites of the neuronal cytoskeleton."

1991 Fiscal Year Annual Research Report

細胞骨格の分子講築と機能〜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(医), 教授 (20010085)

Research Products

[Publications] Hirokawa,N.: "Molecular architecture and dynamics of the neuronal cytoskeleton.in Neuronal cytoskeleton." Wily Liss,Inc.,New York. 5-74 (1991)

[Publications] Okabe,s.: "Actin dynamics in growth cones." J.of Neuroscience. 11. 1918-1929 (1991)

[Publications] Hirokawa,N.: "Kinesin associates with anterogradely transported membraous organelles." J.of Cell biology. 114. 259-302 (1991)

[Publications] Nakata,T.: "Predominant and developmentally regulated expression of dynamin in neurons." Neuron. 7. 461-469 (1991)

[Publications] Takemura,R.: "Developmental expression of tau mRNAs in rat brain." Neuroscience. 44. 393-407 (1991)

[Publications] Takeuchi,M.: "The 72KD microtubule associated protein from porcine brain." J.of Neurochemistry. (1992)

[Publications] Nakata,T.: "Organization of cortical cytoskeleton of cultured chromaffin cells and involvement in secretion as revealed by quick-freeze,deep-etching,and double-label immunoelectron microscopy." J.of Neuroscience. (1992)

[Publications] Okabe,S.: "Differential behavior of photoactivated microtubules in qrowing axon of mouse and frog neurons." J.of Cell Biology,. (1992)

[Publications] Kanai,Y.: "Microtubule bundling by tau proteins in vivo:analysis of the functional domains."

[Publications] Sato-Yoshitake,R.: "Phosphorylation of kinesin regulates its binding to synaptic vesicles."

[Publications] Takemura,R.: "Reorganization of brain spectrin (fodrin) during differentiation of PC12 cells."

[Publications] Okada,Y.: "Functional differentiation of axonal microtubules;Tyrosinated alpha-tubulin rich microtubules for transport,and detyrosinated alpha-tubulin rich microtubules for architecture."

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

[Publications] Nakata,T.: "T-dynamin,a novel member of dynamin-family protein is expressed specific to testis."

[Publications] Maeda,K.: "Dynamin:its enzymatic activity and interaction with microtubules."

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

[Publications] Hirokawa,N.: "Selective stabilization of microinjected tau in the axon and microtubule associated proteins 2 (MAP2) in dendrites of the neuronal cytoskeleton."

廣川信隆東京大学, 医学部(医), 教授 (20010085)