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signal transduction in cells analyzed by imaging and protein rotation

Research Project

Project/Area Number 07458165
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biophysics
Research InstitutionThe University of Tokyo

Principal Investigator

KAWATO Suguru  The University of Tokyo, Graduate School of Arts and Sciences Professor, 大学院・総合文化研究科, 教授 (50169736)

Co-Investigator(Kenkyū-buntansha) OHTA Yoshihiro  The University of Tokyo, Graduate School of Arts and Sciences, Assistant, 大学院・総合文化研究科, 助手 (10223843)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 1996: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1995: ¥4,800,000 (Direct Cost: ¥4,800,000)
Keywordscalcium signaling / glial cells / adrenocortical cells / endosomes / カルシウムオシレーション / 副腎皮質束状層細胞 / ACTH / ステロイドホルモン
Research Abstract

We have examined Ca^<2+> signaling, LDL uptake and the activity of cytochrome P450scc in individual rat brain glial cells and bovine adrenocortical fasciculata cells in order to investigate the molecular mechanisms of steroid hormonc synthesis. With fluorescence video-enhanced microscopy imaging, we have demonstrated that the Ca^<2+> signaling occurred in individual Calcium Green-1 loaded cells upon stimulation with neurotransmitters (scrotonin, glutamate, histamine) to astroglial cells and adrenocorticotropin (ACTH) to adrenal cells at physiological concentration. We observed three patterns of Ca^<2+> signaling which were 1)Ca^<2+> oscillations, 2)step increase in Ca^<2+> concentration, and 3)Ca^<2+> oscillations superimposed on a step-like increase in Ca^<2+>. The results indicated that the Ca^<2+> signaling is the second messenger.
Traffic of cndosomes which contain DiI labeled LDL in astroglial cells and adrcnal cells was observed in the time scale of 30 min with video-cnhanced microscopy Some localization of LDL in the cytoplasm was observed for astrocytes. For oligodendrocytes, incorporated LDL was distributed over the entire cytoplasmic region of both cell body and multiple branched cell processes.
confocal laser scanning microscopy.
Confocal imaging of the activity of cytochrome P450scc was performed in adrenocortical cells. Upon conversion of cholesterol-resorufin by P450scc to pregncnolonc and resorufin, the P450scc activity was observed by the strong fluorescence spots and weak fluorescence patches of resorufin.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (19 results)

All Other

All Publications (19 results)

  • [Publications] 川戸 佳: "膜蛋白質のダイナミクスと機能" 日本膜学会. 21. 240-246 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kimoto,T.: "Adrenoconicouopin induces Calcium Oscillations in Adrenal Fasciculata Cells : Single Cell Imaging" Biochem.Biophys.Res.Comm.221. 25-32 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kimoto,T.: "Digital Fluorescence Imaging of Elementary Steps of Neurosteroid Synlhesis in Rat Brain Glial Cells" J.Pharm.Biomed.Anal.(印刷中). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Homma,R: "Time-resolved Study of Effect of Chlorpromazine on Mobility of Cytochrome P450 and Phospholipids in the Inner Membrane of Adrenocortical Mitochondria" J.Pharm.Biomed.Anal.(印刷中). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] KImoto,T.: "Imaging of Calcium Oscillations and Activity of Cytochrome P450scc in Adrenocortical Cells" Oxygen Homeostasis and and Its Dynamics. (印刷中). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 川戸 佳: "活動する脳グリア細胞のイメージング" 蛋白質核酸酵素. 42(表紙). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kimoto, T.: "Adrenocorticotropin induces Calcium oscillations in Adrenal Fasciculata Cells : Signale Cell Imaging" Biochem.Biophys.Res.Comm.221. 25-32 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kimoto, T.: "Digital Fluorescence Imaging of Elementary Steps of Neurosteroid Synthesis in Rat Brain Glial Cells" J.Pharm.Biomed.Anal.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Homma, R: "time-resolved Study of Effect of Chlorpromazine on Mobility of Cytochrome P450 and Phospholipids in the Inner Membrane of Adrenocortical Mitochondria" J.Pharm.Biomed.Anal.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Kimoto, T.: "Imaging of Calcium Oscillations and Activity of Cytochrome P450scc in Adrenocortical Cells" Oxygen Homeostasis and and Its Dynamics. (in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 川戸 佳: "膜蛋白質のダイナミクスと機能" 日本膜学会. 21. 240-246 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kimoto,T.: "Adrencoricotropin induces Calcium Oscillations in Adrenal Fasciculata Cells:Single Cell Imaging" Biochem.Biophys.Res Comm.221. 25-32 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kimoto,T.: "Digital Fluorescence Imaging of Elementary Steps of Neurosteroid Synthesis in Rat Brain Glial Cells" J.Pharm.Biomed.Anal.(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Homma,R.: "Time-resolved Study of Effect of Chlorpromazine on Mobility of Cytochrome P450 and Phospholopids in the Inner Menbrane of Adrenocorical Mitochondria" J.Pharm.Biomed.Anal.(印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Kimoto,T.: "Imaging of Calcium Oscillations and Activity of Cytochrome P450scc in Adrenocortical Cells" Oxygen Homeostasis and and Its Dynamics. (印刷中). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 川戸 佳: "活動する脳グリア細胞のイメージング" 蛋白質核酸酵素. 42. (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] 川戸佳: "シトクロムP450の膜中での運動と細胞応答・信号伝達" 蛋白質核酸酵素. 40. 43-51 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Yamada.M.: "Dynamic Interactions of Rabbit Liver Cytochrome P450IA2 and P45011B4 with Cytochrome b5 and NADPH-Chtochrome P450 Reductase in Proteoliposomes" Biochemistry. 345. 10113-10119 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Kimoto,T.: "Multi-view Imaging of Calcium Signaling and distribution of Steroidogenic Enzymes in Adrenal Fasciculate Cells" 2nd Hamamatsu Symposium on Cell-Cell Communications. (印刷中). (1996)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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