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Molecular mechanisms of the synapse formation and maintenance in central nervous system

Research Project

Project/Area Number 03454154
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field General medical chemistry
Research InstitutionTOKYO METROPOLITAN INSTITUTE FOR NEUROSCIENCES

Principal Investigator

KURODA Yoichiro  TOKYO METOROPOLITAN INSTITUTE FOR NEUROSCIENCES, 神経生化学研究部門, 副参事研究員 (30073084)

Co-Investigator(Kenkyū-buntansha) KOBAYASHI Kazuo  東京都神経科学総合研究所, 神経生化学研究部門, 主事研究員 (80100139)
ICHIKAWA Masumi  東京都神経科学総合研究所, 解剖発生研究部門, 主事研究員 (20124414)
Project Period (FY) 1991 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 1993: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1992: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1991: ¥3,400,000 (Direct Cost: ¥3,400,000)
KeywordsSynapse formation / Cultured neuron / Asaay system / Intracellular Ca^<2+> / Protein kinase / Ganglioside / Process / Central nervous system / タンパク質リン酸化 / エクト・プロテインキナーゼ / 大脳皮質 / カルシュウム / エンドグルコセラミダーゼ / 海馬 / 培養ニュ-ロン / プロテインキナ-ゼ / ATP
Research Abstract

Synapse formation between cultured cerebral cortical neurons was observed by multi-site Ca^<2+> fluorometry(Muramoto, K., et al., Proc.Japan Acad.Ser.B, 64,319,1988). Most neurons in the culture showed synchronous oscillations of the fluorescence intensity which correspond to spontaneous synaptic activity. Continuous application of K-252b, a protein kinase inhibitor, blocked the synchronous firing and significantly decreased the number of synapses identified morphologically using electron microscopy (Kuroda, Y., et al., Neurosci.Lett., 135,255,1992). Since K-252b does not permeate the cell membrane, the data strongly suggest that phosphorylation of cell surface by an ecto-protein kinase (Ehrlich, Y.H., et al., Nature, 320,67,1986) is inhibited by K-252b. Since it has been reported that an ecto-protein kinase activity is regulated by a specific type of gangliosides (Tsuji, S., et al., J.Biochem., 104,498,1988), we applied purified endoglycoceramidase (EGCase) which can remove all carbohydrate moieties from any ganglioside but not from proteins and others (Ito, M., et al., J.Biol.Chem., 264,9510,1989) to the cortical culture. Continuous presence of EGCase together with its activator protein inhibited the occurrence of synchronous oscillation after 8 days in culture, when control culture showed oscillation which represents a significant amount of synapse formation between cerebral cortical neurons. The inhibition was dose-dependent and without any significant morphological changes of neurons and their neurites which was observed by phase-contrast microscopy and immunostaining of MAP 2 and neurofilament.

Report

(4 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • 1991 Annual Research Report
  • Research Products

    (28 results)

All Other

All Publications (28 results)

  • [Publications] Muramoto,K.: "Frequency of synchronous oscillations of neuronal activity increases during development and is correlated to the number of synapses in culture cortical neuron networks" Neuroscience Letters. 163. 163-165 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Ichikawa,M.: "Formation and maturation of synapses in primary cultures of rat cerebral cortical cells:an electron microscopic study" Neuroscience Research. 16. 95-103 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kuroda,Y.: "Block of synapse formation between cerebral cortical neurons by a protein kinase inhibitor" Neuroscience Letters. 135. 255-258 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 村本和世: "ニューロン間のネットワーク形成における糖鎖の役割" 蛋白質核酸酵素. 37. 1894-1901 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 黒田洋一郎: "シナプスの死、ニューロンの死-アルツハイマー病の原因・発症機序をめぐって" 実験医学. 10. 54-57 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Ichikawa,M.: "Expression of synaptophysin during synapse formation between dissociated cortical neurons" Neuroscience Research. 12. 452-458 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 黒田洋一郎: "『ボケの原因を探る』" 岩波新書, 223 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Muramoto, K.: "Frequency of synchronous oscillations of neuronal activity increases during development and is correlated to the number of synapses in culture cortical neuron networks" Neuroscience Letters. 163. 163-165 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Ichikawa, M.: "Formation and maturation of synapses in primary cultures of rat cerebral cortical cells : an electron microscopic study" Neuroscience Research. 16. 95-103 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Kuroda, Y.: "Block of synapse formation between cerebral cortical neurons by a protein kinase inhibitor" Neuroscience Letters. 135. 255-258 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Ichikawa, M.: "Expression of syanptophysin during synapse formation between dissociated cortical neurons" Neuroscience Research. 12. 452-458 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Muramoto,K.: "Frequency of synchronous oscillations of neuronal activity increases during development and is correlated to the number of synapses in culture cortical neuron networks" Neuroscience Letters. 163. 163-165 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Ichikawa,M.: "Formation and maturation of synapses in primary cultures of rat cerebral cortical cells:an electron microscopic study" Neuroscience Research. 16. 95-103 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Kuroda,Y.: "Block of synapse formation between cerebal cortical neurons by a protein kinase inhibitor." Neuroscience Letters. 135. 255-258 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 黒田 洋一郎: "記憶と忘却の脳内メカニズム" BIOmedica. 7. 698-707 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kazuyo MURAMOTO: "Evidence for that gangliosides are essential for synapse formation between cerebral cortical neurons" Society for Neuroscience Abstracts. 18. 1113- (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kazuo KOBAYASHI: "Characterization of synapse formation between CNS neurons in culture" Society for Neuroscience Abstracts. 18. 1113- (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 村本 和世: "ニューロン間のネットワーク形成における糖鎖の役割" 蛋白質核酸酵素. 37. 1894-1901 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.P.C.Robinson: "Characterization of Spontaneous Synchronized Calcium Transients in Cultured Networks of Cerebral Cortical Neurons." J.Neurophysiol.(1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 黒田 洋一郎: "ボケの原因を探る" 岩波新書(岩波書店), 223 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Kuroda,Y.,Ichikawa,M.,Muramoto,K.,Kobayashi,K.,Matsuda,Y.,Ogura,A.,Kudo,Y.: "Block of syn apse formation between cerebal cortical neurons by a protein kinase inhibitor." Neuroscience Letters. 135. 255-258 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kuroda.Y.: "Rapid consolidation of assciations between multi-modal memories by intrahippoc ampal connections through cortico-hippocampal closed circuit." Concepts In Neuroscience 2.(1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Sekino,Y.,Ito,K-I.,Miyakawa,H.,Kato,H.,Kuroda,Y.: "Adenosine(A_2)antagonist inhibits induction of long-term potentiation of evoked synaptic potentials but not of the population spike in hippocampal CA-1 neurons." Bilchem.Biophys.Res.Comm.,. 181. 1010-1014 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kobayashi,K.,Sawada,H.& Hayashi,M.,Ishikawa,M.,Kuroda,Y.,: "Idetiication of a brai n-specific 27/26-kDa extracellular protein with the monoclonal antibody to differentiated PC12 h pheochromocytoma cells." Experimental Cell Research,. 193. 151-154 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Ichikawa,m.,Kimura-Kuroda,J.,Yasui,K.& Kuroda,Y.: "Expression of synaptophysin during synapse formation bwtween dissociated cortical neurons." Neuroscience Research. 12. 452-458 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kuroda,Y.: "Activity-dependent release of ATP and adenosine derivatives can trigger molecular caseades for the memory process in human brain.Role of adenosine and adenine nucleotides in the biological system.S.Imai & M.Nakazawa,eds.," Chapter 55,605-615,Elsevier Science Publishers BV.(1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 黒田 洋一郎: "脳の機能と遺伝子『脳と遺伝子』" 平凡社215ー224, (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 黒田 洋一郎(伊藤 正男編): "記憶・思考の物質過程ー記憶における“くりかえし"の重要性ー『脳と思考』" 紀ノ国屋書店PP193ー194, (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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