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Mechanisms by which protein kinases modify catecholamine secretion and analyses of them by evanescent-wave fluorescence microscopy

Research Project

Project/Area Number 10670037
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field General physiology
Research InstitutionNIIGATA UNIVERSITY

Principal Investigator

WARASHINA Akira  School of Medicine, NIIGATA UNIVERSITY, Associate Professor, 医学部, 助教授 (50064580)

Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2000: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥1,400,000 (Direct Cost: ¥1,400,000)
Keywordschromaffin cell / catecholamine secretion / protein kinase C / nicotine / muscarine / evanescent-wave microscope / acridine orange / exocytosis / ラット副腎髄質細胞 / wortmannin / LY294002 / PI3-キナーゼ / プロテイン キナーゼC / 分泌顆粒 / ラット副腎髄質 / プロテインキナーゼA / プロテインキナーゼC / ホルボルジブチレイト / ホルスコリン / ミオシン軽鎖キナーゼ
Research Abstract

Effects of the activation of different types of protein kinases in adrenal chromaffin cells on secretory responses stimulated by various secretagogues were analyzed. The following results were obtained.
1. Ca^<2+>-indicator-loaded chromaffin cells in the perfused rat adrenal medulla were stimulated by various stimulants and changes in [Ca^<2+>]_i (Ca-response) and in catecholamine secretion (secretory response) were measured simultaneously. On activating protein kinase C (PKC) with phorbol dibutyrate, secretory processes were affected through at least, two different pathways. One is to inhibit the early-stage of secretory processes stimulated by muscarine, bradykinin and histamine. This inhibition was associated with a reduction of Ca^<2+> entry mediated by the receptor/G-protein. The other is to promote the late-stage of secretory processes occurring after an increase in [Ca^<2+>]_i. This promotion involves any kind of secretagogues including nicotine and the receptor agonists mentione … More d above. The activation of PKA with forskolin promoted the late-stage of secretory processes in a similar manner to PKC.Although wortmannin and LY29004 have been reported to suppress catecholamine secretion by inhibition of Pl3-kinase, these agents may reduce Ca^<2+> entry through their nonspecific effects on Ca^<2+> channels.
2. To investigate the mechanism by which PKC promote the late-stage secretory processes, the evanescent-wave fluorescence microscopy was applied to visualize secretory granules near plasma membrane of chromaffin cells. Some of acridine orange-stained granules were disappeared when cells were stimulated by electrical stimuli, indicating that observed granules were docked on the membrane to prepare exocytotic secretion. It was found in a cooperative study with Dr.Terakawa at Hamamatsu Medical School that the activation of PKC increases the number of granules docking on the plasma membrane. This result may provide one of possible mechanisms by which PKC promotes the late-stage of secretory processes. Less

Report

(4 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] Warashina,A.: "Modulation of early and late secretory responses by activation of protein kinases in the rat adrenal medulla."Biological Signals and Receptors. 7. 307-320 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 藁科彬: "分泌刺激とPKC"生体の科学. 49. 261-263 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Warashina,A.: "Light-evoked recovery from wortmannin-induced inhibition of catecholamine secretion and synapse transmission."Archives of Biochemistry and Biophysics. 367. 303-310 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Warashina,A.: "Mechanism of wortmannin-induced in hibition of secretory responses in rat adrenal medullary cells."Life Sciences. 67. 2587-2593 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Warashina,A.: "Mechanism by which wortmannin and LY294002 inhibit catecholamine secretion in rat adrenal medullary cells"Cell Calcium. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Fujita,T.: "The Adrenal Medulla"Hokkaodo University Press (Edited by Kanno,T. et al.). 394-(9) (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Warashina, A.: "Modulation of early and late secretory responses by activation of protein kinases in the rat adrenal medulla."Biological Signals and Receptors. 7. 307-320 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Warashina, A.: "Bunpitu sigeki to PKC"Seitai no Kagaku. 49(in Japanese). 261-263 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Warashina, A.: "Light-evoked recovery from wortmannin-induced catecholamine secretion and synaptic transmission."Archives of Biochemistry and Biophysics. 367. 303-310 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Warashina, A.: "Mechanism of wortmannin-induced inhibition of secretory responses in rat adrenal medullary cells."Life Sciences. 67. 2587-2593 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Warashina, A.: "Mechanism by which wortmannin and LY-294002 inhibit catecholamine secretion in rat adrenal medullary eells."Cell Calcium. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Fujita, T., et al.: "The Adrenal Medulla"Hokkaido University Press. 9/394 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Warashina,A.: "Mechanism of wortmannin-induced inhibition of secretory responses in rat adrenal medullary cells."Life Sciences. 67. 2587-2593 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Warashina,A.: "Mechanism by which wortmannin and LY294002 inhibit catecholamine secretion in rat adrenal medullary cells."Cell Calcium. (in press).

    • Related Report
      2000 Annual Research Report
  • [Publications] Warashina,A: "Light-evoked recovery from wortmannin-induced inhibition of catecholamine secretion and synaptic transmission."Archives of Biochemistry and Biophysics. 367. 303-310 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Warashina,A: "Modulation of early and late secretory processes by activation of protein kinases in the rat adrenal medulla." Biological Signals and Receptors. 7・6. 307-320 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] 藁科 彬: "分泌刺激とPKC" 生体の科学. 49・4. 261-263 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Warashina,A(分担著者): "The Adrenal Chromaffin Cell: Archetype and Exemplar of Cellular Signaling in Secretory Control." Edited by Kannno,T.,Nakazato,Y.& Kumakura,K.Hokkaido University Press, 169-177 (1998)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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