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ANALYSIS OF MOLECULAR MECHANISMS FOR NEURONAL POLARITY AND AXON FORMATION BY CRMP-2

Research Project

Project/Area Number 13680872
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Neuroscience in general
Research InstitutionNARA INSTITUTE OF SCIENCE AND TECHNOLOGY

Principal Investigator

INAGAKI Naoyuki  Nara Institute of Science and Technology, Division of Signal Transaction, Associate Professor, バイオサイエンス研究科, 助教授 (20223216)

Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
KeywordsNeuron / Axon / Dendrite / Cell Polarity / CRMP-2 / Tublin / Microtubule / Proteome / プロテオーム
Research Abstract

Identification of CRMP-2 as a molecule involved in neuronal polarity and axon formation
Formation of an axon and dendrites is a fundamental step in neuronal development. In this research, we found that CRMP-2 was enriched in growing axons of cultured hippocampal neurons. Overexpression of CRMP-2 in the cells led to the formation of supernumerary axons without remarkable increase in the total number of neurites. CRMP-2 not only enhanced the formation of multiple axons during the initial stages of axon specification but also induced axon sprouting from already formed dendrites. Furthermore, expression of truncated CRMP-2 mutants suppressed the formation of primary axon in a dominant negative manner. Thus, local concentrations of CRMP-2 in neurites appear to play a critical role in axon induction of hippocampal neurons, thereby establishing and maintaining neuronal polarity.
In order to examine the molecular mechanisms for CRMP-2 induced axonal formation, we also investigated the CRMP-2 bin … More ding proteins. Here, we identified two activities of CRMP-2: tublin-heterodimer binding and the promotion of microtuble assembly. CRMP-2 bound tublin dimers with higher affinity than it bound microtubules. Association of CRMP-2 with microtubules was enhanced by tublin polymerization in the presence of CRMP-2. The binding property of CRMP-2 with tubulin was apparently distinct from that of Tau, which preferentially bound microtubules. In neurons, overexpression of CRMP-2 promoted axonal growth and branching. A mutant of CRMP-2, lacking the region responsible for microtuble assembly, inhibited axonal growth and branching in a dominant-negative manner. Taken together, our results suggest that CRMP-2 regulate axonal growth and branching as a partner of the tublin hetereodimer, in a different fashion from traditional MAPs.
Search for molecules involved in neuronal polarity and axon formation by protemic strategy
We further started to investigated the molecules involved in axon and polarity formation in addition to CRMP-2 by proteomic strategy. In order to searched molecules involved in axon and polarity formation, we established a 2-DE two-dimensional gel electrophoresis (2-DE) system of highresolution. We utilized fourteen large 2-DE gels (twelve 24 cm x 70 cm gels and two 18 cm x 70 cm gels) which were assembled into a 93 cm x 103 cm cybergel. Our data suggested that the cyber gel can display more than 11,000 protein spots expressed in a 1-10^5 dynamic range in cells. The utilization of present strategy led to about 500% increase in the number of detected spots in comparison to a standard procedure. With this system, we are currently screening molecules which are expressed during the processes of neuronal polarity formation. Less

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (40 results)

All Other

All Publications (40 results)

  • [Publications] Inagaki, N. et al.: "CRMP-2 induces Axons in Cultured Hippocampal Neurons"Nature Neurosci.. 4. 872-873 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Canossa, M., et al.: "Regulated secretion of neurotrophins by metabotropic glutamate group I (mGluRI)-and Trk-receptor activation is mediated via phospholipase C signaling pathways"EMBO J.. 20. 1641-1650 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Taya, S., et al.: "Direct Interaction of Insulin-like growth factor-1 receptor with leukemia-associated RhoGEF"J.Cell Biol.. 155. 809-819 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Oguri: "Proteome analysis of rat hippocampal neurons by multiple large gel two-dimensional electrophoresis"Proteomics. 2. 666-672 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] N.Inagaki, et al.: "High resolution large gel two-dimensional electrophoresis for proteomics"BIO forum Int.. 6. 324-325 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y.Fukuta, et al.: "CRMP-2 binds to tublin heterodimers to promote microtubule assembly"Nature Cell Biol.. 4. 583-591 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Inagaki, N., et al.: "Electrophorese bidimension nelle haute sur gel extra large pour la proteomique"BIOforun Europe Edition Francaise. 1. 28-29 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 稲垣直之, 貝淵弘三: "CRMP-2は海馬神経細胞の軸索形成を誘導する"細胞工学. 20. 1408-1409 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 深田優子, 稲垣直之, 貝淵弘三: "神経細胞の極性:神経軸索と樹状突起の運命決定"細胞工学. 52. 230-234 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 有村奈利子, 稲垣直之, 貝淵弘三: "CRMP-2による神経極性形成の制御機構"実験医学. 17. 2309-2311 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 稲垣直之, 貝淵弘三: "神経細胞の軸索および極性形成の分子機構"生体の科学. 52. 230-234 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 稲垣直之: "高解像度ラージゲルを用いた二次元電気泳動法による細胞内発現蛋白質の網羅的検出"実験医学. 20. 85-88 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 稲垣直之: "細胞内の空間シグナル"細胞工学. 21. 345 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 稲垣直之: "ポストシナプス・樹状突起スパインにおけるCaMKIIの空間的シグナリング"生体の科学. 54. 90-96 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 勝田和大, 野村英子, 稲垣直之: "Multiple large gel two-dimensional electrophoresis for proteomics"Jap.J.Electrophresis. (in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 稲垣直之, 稲垣昌樹: "神経ネットワークとシナプスダイナミックス"塩坂貞夫編、金芳堂(in press). (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Inagaki, N., Chikara, K., Arimura, N., Menager, C., Kawamoto, Y., Matsuo, N., Nishimura, T., Amano, M., and Kaibuchi, K: "CRMP-2 Induces Axons in Cultured Hippocampal Neurons"Nature Neurosci.. 4. 781-782 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Cannosa, M., Gartner, A., Campana, G., Inagaki, N., and Thoenen, H.: "Regulated secretion of neurotrophins by metabotropic glutamate group l (mGluRl)- and Trk-receptor activation is mediated via phospholipase C signaling pathway"EMBO J.. 20. 1641-1650 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Taya, S., Inagaki, N., Sengiku, H., Makino, H., Iwamatsu, A., Urakawa, I., Nagano, K., Kataoka, S., and Kaibuchi, K.: "Direct Interaction of Insulin-like growth factor-1 receptor with leukemia-associated RhoGEF"J. Cell Bio.. 155. 809-819 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Oguri, T., Takahata, I., Katsuta, K., Nomura, E., Hidaka, M., and Inagaki, N.: "Proteome analysis of rat hippocampal neurons by multiple large gel two-dimensional electrophoresis"Proteomics. 2. 666-672 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Fukata, Y., Itoh, T.J., Kimura, T., Menager, C., Nishimura, T., Shiromizu, T., Watanabe, H., Inagaki, N., Iwamatsu, A., Hotani, H., Kaibuchi, K.: "CRMP-2 binds to tubulin heterodimers to promote microtubule assembly"Nature Cell Biol.. 4. 583-591 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Inagaki, N., Katsuta, K., Nomura, E., Ueda, T., Toriyama, M., and Mori, T.: "High resolution large gel two-dimensional electrophoresis for proteomics"BIOforun Int.. 6. 324-325 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Inagaki, N., Katsuta, K., Nomura, E., Ueda, T., Toriyama, M., and Mori, T.: "Electrophorese bidimensionnelle haute resolution sur gel extra large pour la proteomique"BIOforun Europe Edition Franchise. 1. 28-29 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Oguri: "Proteome analysis of rat hippocampal neurons by multiple large gel two-dimensional electrophoresis"Proteomics. 2. 666-672 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] N.Inagaki: "High resolution large gel two-dimensional electrophoresis for proteomics"BIO forum Int.. 6. 324-325 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Y.Fukata: "CRMP-2 binds to tubulin heterodimers to promote microtubule assembly"Nature Cell Biol.. 4. 583-591 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 稲垣直之: "細胞内の空間シグナル"細胞工学. 21・4. 345 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 勝田和大: "Multiple large gel two-dimensional electrophoresis for proteomics"Jap. J. Electrophresis. (In press). (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 稲垣直之: "高解像度ラージゲルを用いた二次元電気泳動法による細胞内発現蛋白質の網羅的検出"実験医学. 21・1. 85-88 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 稲垣直之: "ポストシナプス・樹状突起スパインにおけるCaM KIIの空間的シグナリング"生体の科学. 54・2(In press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 稲垣直之: "神経ネットワークとシナプスダイナミックス"塩坂貞夫編、金芳堂(In press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Canossa, M., et al.: "Regulated secretion of neurotrophins by metabotropic glutamate group I(mGluRI)-and Trk-receptor activation is mediated via phospholipase C signaling pathways"EMBO J.. 20. 1641-1650 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Inagaki, N., et al.: "CRMP-2 induces Axons in Cultured Hippocampal Neurons"Nature Neurosci.. 4. 872-873 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Taya, S., et al.: "Direct Interaction of Insulin-like growth factor-1 receptor with leukemia-associated RhoGEF"J.Cell Biol.. 155. 809-819 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Oguri, T., et al.: "Proteome analysis of rat hippocampal neurons by multiple large gel two-dimensional electrophoresis"Proteomics. (印刷中). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] 稲垣直之, 貝淵弘三: "CRMP-2は海馬神経細胞の軸索形成を誘導する"細胞工学. 20(10). 1408-1409 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 深田優子, 稲垣直之, 貝淵弘三: "神経細胞の極性:神経軸索と樹状突起の運命決定"細胞工学. 52(3). 230-234 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 有村奈利子, 稲垣直之, 貝淵弘三: "CRMP-2による神経極性形成の制御機構"実験医学. 17(11). 2309-2311 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 稲垣直之: "高解像度ラージゲルを用いた二次元電気泳動法による細胞内発現蛋白質の網羅的検出"実験医学. 21(1). 85-88 (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] 稲垣直之, 貝淵弘三: "神経細胞の軸索および極成形性の分子機構"生体の科学. 52(3). 230-234 (2001)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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