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Ultracytochemical study of gap junctions under low electron beam damage condition

Research Project

Project/Area Number 61440024
Research Category

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

Allocation TypeSingle-year Grants
Research Field General anatomy (including Histology/Embryology)
Research InstitutionKyoto University

Principal Investigator

OGAWA Kazuo  Kyoto University, Faculty of Medicine, Professor, 医学部, 教授 (20077556)

Co-Investigator(Kenkyū-buntansha) NODA Toru  Kyoto University, Faculty of Medicine, Assistant, 医学部, 助手 (50156204)
SAKAI Masahiro  Kyoto University, Faculty of Medicine, Assistant, 医学部, 助手 (40183363)
FUJIMOTO Kazushi  Kyoto University, Faculty of Medicine, Lecturer, 医学部, 講師 (50159125)
FUJIMOTO Toyoshi  Kyoto University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (50115929)
Project Period (FY) 1986 – 1988
Project Status Completed (Fiscal Year 1988)
Budget Amount *help
¥31,000,000 (Direct Cost: ¥31,000,000)
Fiscal Year 1988: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1987: ¥10,000,000 (Direct Cost: ¥10,000,000)
Fiscal Year 1986: ¥20,000,000 (Direct Cost: ¥20,000,000)
Keywordsgap junction / calmodulin / cryo-electron microscope / Ca^<++>-ATPase / コネクソン / Ca^<++>ーATPase活性 / クライオトランスファーホルダー / 低電子線損傷 / 凍結超薄切片方法 / 細胞分画法 / 電顕細胞化学 / カルモジュリン
Research Abstract

In this project, the ultrastructure of gap junctions which play an important role in cell-to cell communication was investigated by using ultracytochemical thchnique under a cryo-electron microscope. The data we obtained are as follows:
1. By an ultracytochemical method that we developed to detect the intracellular localization of calmodulin (one of the calcium-binding proteins) binding sites, it was demonstrated that rat liver and heart gap junctions have calmodulin binding sites on their cytoplasmic surface, suggesting that the function of gap junctions may be regulated by calcium ions.
2. The cryo-electron microscopic observation of negatively-stained isolated gap junctions revealed a quite different image as compared with that observed under a conventional electron microscope in terms of the connexon distribution pattern and the average diameter of connexons.
3. The localization of Ca^<++>-ATPase on isolated heart gap junctions under a cryoelectron microscope was found around connexon particles with a very precise and uniform distribution pattern, whereas coarse and aggregated reaction products were observed under a convertional electron microscope. These results indicate that isolated gap junction observation under a cryo-electron microscope can be considered very useful in detecting near-to-native structure.

Report

(4 results)
  • 1988 Annual Research Report   Final Research Report Summary
  • 1987 Annual Research Report
  • 1986 Annual Research Report
  • Research Products

    (28 results)

All Other

All Publications (28 results)

  • [Publications] 酒井眞弘: 解剖学雑誌. 64. (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Fujimoto: Journal of Histochemistry and Cytochemistry. 37. 249-256 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M.Sakai: Acta histochemica et cytochemica. 21. 535-549 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M.Sakai: Journal of Electron Microscopy. 37. 271 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] S.Kondo: Acta histochemica et eytochemica. 21. 521-533 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Fujimoto: Acta histochemica et cytochemica. 20. 341-342 (1987)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T.Tanaka: "Proceeding of the IVth Asia-Pacific Conference and Workshopon Electron Microscopy" V.Mangclavivaj,W.Banchorndhevakul and P.Ingkaninun, 537-538 (1988)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K.Fujimoto: "Electron Microscopy 1986,Vol.III,Biology(I)" T.Imura,S.Maruse and T.Suzuki, 1869-1872 (1986)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K. Fujimoto: "Ultracytochemistry of calmodulin binding sites in myocardial cells by staining of frozen thin sections with colloidal gold-labeled calmodulin." Journal of Histochemistry and Cytochemistry. 37. 249-256 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M. Sakai: "Ultrastructure and Ca^<++>-ATPase activity of gap junctions under cryo-electron microscopic observation" Acta histochemica et cytochemica. 21. 535-549 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] M. Sakai: "Ultrastructure of gap junctions at ultra-low temperatures by using cryo-transfer holder system" Journal of Electron Microscope. 37. 271 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] S. Kondo: "Calcium-activated adenosine triphosphatase and gap junctions in rat epidermis" Acta histochemica et cytochemica. 21. 521-533 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] K. Fujimoto: "Cytochemistry of calmodulin-binding sites in myocardial cells by staining frozen thin-sections with colloidal gold labeled calmodulin" Acta histochemica et cytochemica. 20. 341-342 (1987)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] T. Tanaka: Proceeding of the IVth Asia-Pacific Conference and Workshop on Electron Microscopy. V. Mangclaviraj, W. Banchorndhevakul and P. Ingkaninun, 537-538 (1988)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1988 Final Research Report Summary
  • [Publications] 酒井眞弘: 解剖学雑誌. 64. (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] K.Fujimoto: Journal of Histochemistry and Cytochemistry. 37. 249-256 (1989)

    • Related Report
      1988 Annual Research Report
  • [Publications] M.Sakai: Acta histochemica et cytochemica. 21. 535-549 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] M.Sakai: Journal of Electron Microscopy. 37. 271 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] S.Kondo: Acta histochemica et cytochemica. 21. 521-533 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] K.Fujimoto: Acta histochemica et cytochemica. 20. 341-342 (1987)

    • Related Report
      1988 Annual Research Report
  • [Publications] T.Tanaka: "Proceedings of the IVth Asia-Pacific Conference and Workshop on Electron Microscopy" V.Mangclaviraj,W.Banchorndhevakul and P.Ingkaninun, 537-538 (1988)

    • Related Report
      1988 Annual Research Report
  • [Publications] K.Fujimoto: "Electron Microscopy 1986,vol.III,Biology(I)" T.Imura,S.Maruse and T.Suzuki, 1869-1872 (1986)

    • Related Report
      1988 Annual Research Report
  • [Publications] K. Fuzimoto: Proc., XIth, Int., Cong., on Electron Microrcopy, Kyoto. 3. 1869-1872 (1986)

    • Related Report
      1987 Annual Research Report
  • [Publications] K. Fuzimoto: J. Histochem. Cytochem.

    • Related Report
      1987 Annual Research Report
  • [Publications] M. Sakai: J. Electron Microsc.37. (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] T. Tanaka: Proc.,4th, Asia-Pacific, Conference on Electron Microscopy, Bargkok. (1988)

    • Related Report
      1987 Annual Research Report
  • [Publications] K.Fujimoto: Proc.【XI】th Int.Cong.on Electron Microscopy,Kyoto. 3. 1869-1872 (1986)

    • Related Report
      1986 Annual Research Report
  • [Publications] K.Fujimoto: J.Histochem.Cytochem.

    • Related Report
      1986 Annual Research Report

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Published: 1987-03-31   Modified: 2016-04-21  

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