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Biochemical Investigation of the Changes of Cytoskeletons during the Exocytosis

Research Project

Project/Area Number 04807119
Research Category

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

Allocation TypeSingle-year Grants
Research Field Obstetrics and gynecology
Research InstitutionOsaka University

Principal Investigator

NISHIZAKI Takamichi  Osaka Univ.Med.School, Assistant Professor, 医学部, 助手 (20237689)

Co-Investigator(Kenkyū-buntansha) MIYAKE Akira  Osaka Univ.Med.School, Lecturer, 医学部, 講師 (90093468)
YOSHIMOTO Yasuhiro  Osaka Univ.Med.School, Assistant Professor, 医学部, 助手 (10182833)
TASAKA Keiichi  Osaka Univ.Med.School, Assistant Professor, 医学部, 助手 (50155058)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1994: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1993: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsExocytosis / Cytoskeleton / Actin / Phalloidin / ファイロジン
Research Abstract

Recent researches on the exocytosis revealed the mechanism of the signal transduction of a lot of kinds of endocrine cells. And the increase to intracellular concentration of calcium is now considered the key step to exocytosis. However the pathway (s) by which elevations of intracellular free calcium stimulate exocytosis remains to be identified. On adrenal chromaffin cells, it was demonstrated that cortical F-actin was decreasing after calcium dependent exocytosis. This study was designed to investigate the changes of cytoskeletons during the exocytosis using rat pituitary cell line, GH_3 cells. We revealed the following results that suggest the changes of cytoskeletons might be important for the exocytosis. First we demonstrated the calcium dependent proinhibit depolymerization of actin filaments, using GH_3 cells that were permeabilized by the treatment of digitonin. Second, we observed GH_3 cells by staining with FITC-phalloidin (which binds to only F-actin) with confocal laser microscopy. At resting stage F-actin was localized only subplasmalemmal region, on the other-hand after daldium stimutation F-actin of subplasmalemmal region was decreasing dramatically and subtle increase of F-actin was observed in plasma region of the cells. These results are coincident with the recent idea that the actin filaments is the essential step to the exocytosis.

Report

(4 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (7 results)

All Other

All Publications (7 results)

  • [Publications] Yamaguchi,M.: "Post-transcriptional inhibitionof mouse placental lactogen-II secretion by transforming growth factor-β1:synergitic effects with epidermal growth factor and interleukin-6" Biol.Reprod.(in Press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kasahara,K.: "High magnesium concentration inhibits ligand-stimulated calcium influx and hormone secretion in rat pituitary lactotropes with involvement of intracellular free magnesium" Biochem,Biophys,Res,Cpmmun.197. 92-99 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Nishizaki,T.: "A Key role for a 145-KDa cytosolic protein in the stimulation of Ca2+ -depend secretion by protein kinase C" J.Biol.Chem.267. 23972-23981 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yamaguchi, M., et al: "Post-transcriptional inhibition of mouse placental lactogen-II secretion by transforming growth factor-beta_1 : synergitic effects with epidermal growth factor and interleukin-6" Biol.Reprod.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kasahara, K., et al: "High magnesium concentration inhibits ligand-stimulated calcium influx and hormone secretion in rat pituitary lactotropes with involvement of intracellular free magnesium" Biochem.Biophys.Res.Commun.197. 92-99 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Nishizaki, T., et al: "A key role for a 145-kDa cytosolic protin in the stimulation of Ca ^<2+>-depend secretion by protein kinase C" J.Biol.Chem.267. 23972-23981 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yamaguchi,M.: "Post-transcriptional inhibition of mouse placental lactogen-II secretion by transforming growtn factor-β1:synergitic effects with epidermal growth factor and interleukin-6" Biol.Reprod.(in press).

    • Related Report
      1994 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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