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1990 Fiscal Year Final Research Report Summary

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
Keywordsgap junction / cryoelectron microscopy / cytochemistry / protein kinase C / phorbol ester / immunoelectron microscopy / connexin 32 / adherent junction
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.

  • Research Products

    (12 results)

All Other

All Publications (12 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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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

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Published: 1993-08-12  

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