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Metabolism and biological activities of lnsP5 and InsP6 in neuronal cells

Research Project

Project/Area Number 09670104
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General pharmacology
Research InstitutionSophia University (1998)
Keio University (1997)

Principal Investigator

SASAKAWA Nobuyuki  Sophia Univ. Faculty of Science and Technology, Associate Prof, 理工学部, 助教授 (20187107)

Co-Investigator(Kenkyū-buntansha) KUMAKURA Konosuke  Sophia Univ. Faculty of Science and Technology, Associate Prof, 理工学部, 教授 (70129790)
Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 1998: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1997: ¥3,200,000 (Direct Cost: ¥3,200,000)
KeywordsInositol polyphosphates / exocytosis / adrenal chromaffin cell / シナプトタグミン
Research Abstract

Several kinds of stimulants are known to induce inositol1,3,4,5,6-pentakisphosphate (InsP5) and inositol hexakisphosphate (InsP6) accumulations in intact adrenal chromaffin cells, N1E-115 cells and rat cerebellar granule cells. Recently, we suggested that the binding of inositol polyphosphates to the C2B domain of synaptotagmin may clamp spontaneous fusion of the docked or primed vesicles in adrenal chromaffin cells. In this study, we investigated the CaィイD12+ィエD1-evoked release of InsP5 and InsP6, and the effects of the antibodies against synaptotagmin C2A and C2B domains in digitonin-permeabilized adrenal chromaffin cells. When the cells were stimulated with CaィイD12+ィエD1 (100μM), transient increase of InsP5 and InsP6 in the cytosolic medium were observed. Upon a CaィイD12+ィエD1-challenge, amount of InsP5 and InsP6 in the cytosolic medium reached the maximum at 15 sec. Anti-C2A antibody (C2A Ab) completely inhibited CaィイD12+ィエD1-induced InsP5 and InsP6 accumulation in cytosolic medium. Anti-C2B antibody (C2B Ab) also inhibited CaィイD12+ィエD1-induced InsP5 and InsP6 accumulation in the cytosolic medium. Since pretreatment with C2B Ab during permeabilization, but not with C2A Ab, induced increase of InsP5 and InsP6 in the permeabilizing buffer, the mechanisms of the inhibitory effects of the C2B Ab is different from those of C2A Ab. These results strongly suggest that the binding of increased intracellular CaィイD12+ィエD1 to the C2A domain liberate InsP5 and InsP6 from the C2B domain of synaptotagmin, supporting our idea of the clamp of fusion by synaptotagmin and inositol polyphosphates.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (22 results)

All Other

All Publications (22 results)

  • [Publications] Sasakawa, et al.: "Increase in cytosolic inositol pentakis and hexakisphosphate--"Japanese J. Pharmacol. 73.suppl.. 226 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ohara-Imaizumi, M. et al.: "Distinct roles of C2A and C2B domains of synaptotagmin in tha--"Proc. Matl. Acad. Sci. USA.. 94. 287-291 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sasakawa, et al.: "Regulation of exocytosis by inositol polyphosphates Via--"Japanese J. Pharmacol. 76.suppl.l. 136 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Misonou, H. et al.: "Protein kinase C controls the priming step of regulatory exocytosis-"Cell. Mol. Neurobiol. 18. 379-390 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 今泉美佳: "開口放出機構におけるシナプトタグミンC2ドメインの役割"生化学、. 70. 1283-1288 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kumakura, K. et al.: "The Adrenal Chromaffin Cell"Hokkaidou Univ. Press ; Kanno, T., Nakazato, Y., and Kumakura, K.(eds). 394 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kumakura, K., et al.: "Keio Univ. Symp. Life Sci. and Med., 2, Neural Development,"Springer : Uemura., Kawamura, K., and Yazaki, T.(eds). 544 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ohara-Imaizumi, M., Fukuda, M., Niinobe, M., Misonou, H., Ikeda, K., Murakami, T., Kawasaki, M., Mikoshiba, K. and Kumakura, K.: "Distinct Roles of C2A and C2B Domains of Synaptotagmin in the Regulation of Exocytosis in Adrenal Chromaffin Cells"Proc. NatL Acad. Sci. USA.. 94. 287-2913 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Misonou, H., Ohara-Imaizumi, M., and Kumakura, K.: "Protein kinase C controls the priming step of regulatory exocytosis in adrenal chromaffin cells"Cell. Mol. Neurobiol. 18. 379-390 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ohara-Imaizumi, M.: "Roles of C2 domains of synaptotagmin in the regulation of exocytosis"Seikagaku. 70. 1283-1288 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Ohara-Imaizumi, M., Kumakura K.: "Roles of inositol polyphosphates in the regulation of exocytosis"Protein, Nucleic acid and Enzyme. 43. ]1789-1793 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kumakura, K., Ohara-Imaizumi, M., Misonou, H.: "Inhibitory clamps of regulated exocytosis and the role of Ca2+ : clamp by synaptotanmin and inositol phosphates. In Kanno, T., Nakazato, Y., and Kumakura, K. (Eds.) The Adrenal Chromaffin Cell"Hokkaido Univ. Press, Sapporo. 199-211, 384 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kumakura, K., Ohara-Imaizumi, M., Sasakawa, N., Fukuda, M., and Ninobe, M., and Mikoshiba, K.: "Roles of synaptotagmin and inositol polyphosphates in the mechanism of exocytosis : The clamp hypothesis. In : Uyemura, K., Kawamura, K. and Yazaki, T. (Eds.) Keio University Symposia for Life Science and Medicine, Vol.2, Neural Development"Springer press. 450-455, 544 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shintani,et al.: "Bioactive 6-Nitronorepinephrine ldentified in mammalian Brain" J.Biol.Chem.271.23. 13561-13565 (1996)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sasakawa,et al.: "lncrease in cytosolic inositol pentakis and hexakisphosphatesーー" Japanese J.Pharmacol.73.suppl.l. 226 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Ohara-lmaizumi,M.et al.: "Distinct roles of C2A and C2B domains of synaptotagmin in theーー" Proc.Natl.Acad.Sci.USA.94. 287-291 (1997)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sasakawa,et al.: "Regulation of exocytosis by inositol polyphosphates via ーーー" Japanese J.Pharmacol.76.suppl.136 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Misonou,H.et al.: "Protein kinase C controls the priming step of regulatory exocytosis -" Cell.Mol.Neurobiol.18. 379-390 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 今泉美佳,他(1998): "イノシトールポリリン酸の役割:開口分泌における調節作用" 蛋白質核酸酵素. 43. 1789-1793 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kumakura,K.et al.: "The Adrenal Chromaffin Cell" Hokkaido Univ.Press;Kanno,T.,Nakazato,Y.,and Kumakura,K.(eds), 394 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Kumakura,K.,et al.: "Keio Univ.Symp.Life Sci.and Med.,2,Neural Development," Springer: Uemura,K.,Kawamura,K.,and Yazaki,T.(eds), 544 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sasakawa,et al.: "Regulation of exocytosis by inositol polyphosphates via synaptotagmin in--" Japanese J.Pharmacol.76.suppl.I. 136P (1998)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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