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Cryoelectron Microscopy and Cytochemistry of Gap Junctions

Research Project

Project/Area Number 01480107
Research Category

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

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

Principal Investigator

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

Co-Investigator(Kenkyū-buntansha) SAKAI Masahiro  Kyoto University, Faculty of Medicine, Department of Anatomy, Assistant, 医学部, 助手 (40183363)
FUJIMOTO Kazushi  Kyoto University, Faculty of Medicine, Department of Anatomy, Lecturer, 医学部, 講師 (50159125)
FUJIMOTO Toyoshi  Kyoto University, Faculty of Medicine, Department of Anatomy, Associate Professo, 医学部, 助教授 (50115929)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1990: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1989: ¥5,800,000 (Direct Cost: ¥5,800,000)
Keywordsgap junction / cryoelectron microscopy / cytochemistry / protein kinase C / phorbol ester / immunoelectron microscopy / connexin 32 / adherent junction / 肝細胞 / 生後変化 / 凍結割断レプリカ / 荷電
Research Abstract

In this project, we have demonstrated the following cytochemical aspects of gap junctions.
1. Enzyme cytochemistry : Ca^<++>-ATPase activity is localized on the cytoplasmic side of gap junctional membranes.
2. Cytochemical demonstration of charge on gap junctions : Cytoplasmic side of gap junctions shows the high density distribution of anionic sites.
3. Immunocytochemistry of Protein Kinase C (PKC) on gap junctions : The phorbol ester, 12-O-Tetradecanoyl Phorbol-13-Acetate (TPA), one of the most potent tumor-promoting agents, is known to inhibit metabolic cooperation between cultured cells. In addition, TPA is also known to activate protein kinase C both in vivo and in vitro. In this project we have demonstrated that TPA-treatment causes a translocation of cytosolic PKC to the gap junction. Our results imply that PKC have a crucial role in TPA-induced inhibition of metabolic cooperation.
4. Immunocytochemistry of gap junction protein (connexin 32) : We have studied the intracellular distribution of connexin 32 in newly born rat hepatocytes. Immunoelectron microscopy demonstrated that the mouse monoclonal antibody against connexin 32 binds directly to the adherent junction region as well as the gap junction in 6 day-old rat hepatocytes. Our findings imply that the adherent junction mediates an early adhesion event between hepatocytes that is a prerequisite for the assembly of connexin 32 of gap junctions.

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (18 results)

All Other

All Publications (18 results)

  • [Publications] Tanaka,T: "Localization of acid phosphatase activty in the lenses of ICR rats." Acta histochem.cytochem.22. 739- (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kitaoka,T: "Distribution of anionic sites in rod photoreceptor cells and pigment epthelial cells revealed by labeling frozen sections with cationized ferritin." Biomedical Res.3. 549-556 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Tanaka,T: "Ultrastructural studies during cataract development in ICR rat lens." Invest.Ophthal.Visual Sci.30. 145- (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kitaoka,T: "Cytochemical demonstration of anionic sites in rod photoreceptor cells and pigment epithelium of rat retinas." J.Histochem.Cytochem.38. 1032- (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] TanaKa,T: "Morphometric analysis of gap junctions in the rat lens during cataract formation." Acta histochem.cytochem.23. 781-792 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Tanaka,T: "Cytochemical localization of acid phosphatase activity in normal and cataractous rat lenses." Acta histochem.cytochem.23. 793-804 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Tanaka, T., Sakai, M. and Ogawa, K.: "Localization of acid phosphatase activity in the lenses of ICR rats." Acta histochem. cytochem.22. 739 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kitaoka, T., Fujimoto, K. and Ogawa, K.: "Distribution of anionic sites in rod photoreceptor cells and pigment epithelial cells revealed by labeling frozen thin sections with cationized ferritin." Biomedical Research. 3. 549-556 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Tanaka, T., Sakai, M., Ihara, N., Ueno, S., Honda, Y. and Ogawa, K.: "Ultrastructural studies during cataract development in ICR rat lens." Invest. Ophthal. Visual Sci.30. 145 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kitaoka, T., Fujimoto, K,. Honda, Y. and Ogawa, K.: "Cytochemical demonstration of anionic sites in rod photoreceptor cells and pigment epithelium of rat retinas." J. Histochem. Cytochem.38. 1032 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Tanaka, T., Sakai, M., Fujimoto, K. and Ogawa, K.: "Morphometric analysis of gap junctions in the rat lens during cataract formation." Acta histochem. cytochem.23. 781-792 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Tanaka, T., Sakai, M., and Ogawa, K.: "Cytochemical localization of acid phosphatase activity in normal and cataractous rat lenses." Acta histochem. cytochem.23. 793-804 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kitaoka,T: "Cytochemical demonstration of anionic sites in rod photoreceptor cells and pigment epithelium of rat retinas." J.Histochem.Cytochem.38. 1032 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Tanaka,T: "Morphomtric analysis of gap junctions in the rat lens during cataract formation." Acta histochem.cytochem.23. 781-792 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Tanaka,T: "Cytochemical localization of acid phosphatase activity in normal and cataractous rat lenses." Acta histochem.cytochem.23. 793-804 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 藤本 和: "肝細胞ギャップ結合の生後発達について" 1990年度生理学研究所研究会ギャップ結合研究会(3). 4 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 藤本和,酒井眞弘,小川和朗: "ギャップ結合の細胞化学" 生体の科学. 40. 645-649 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Kitaoka,T.,Fujimoto,K.and Ogawa,K: "Distribution of anionic sites in rod photoreceptor cells and pigment epithelial cells revealed by labeling frozen thin sections with cationized ferritin" Biomedical Research. 10. (1989)

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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