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1995 Fiscal Year Final Research Report Summary

Purification and characterization of switching factor in hepatocyte signaling and its regulation

Research Project

Project/Area Number 06670158
Research Category

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

Allocation TypeSingle-year Grants
Research Field General medical chemistry
Research InstitutionThe Tokyo Metropolitan Institute of Medical Science

Principal Investigator

KAJIYAMA Yasuo  The Tokyo Metropolitan Insitute of Medical Science Dept.of Biochemical Cell Research Researcher, 生命情報・研究部門, 研究員 (70250214)

Project Period (FY) 1994 – 1995
KeywordsHepatocytes / Adhesion / Glycoconjugates / Ganglioside GM3
Research Abstract

alpha_1-Adrenergic receptor-mediated response are overwhelming, or alpha_1-responses are dominant, in adult rat hepatocytes. Inversely, beta-responses are predominant over alpha1-responses in hepatocytes that have been cultured at a low-cell density (Adrenergic response "alpha*beta exchange"). We found "alpha*beta exchange" was not previously induced in cell-to-cell contact and we examined the adhesion of hepatocytes to terminal galactoside in detail during this project. As a result, it was suggested that adult hepatocytes were bound to terminal galactosides without binding specificity, and then alpha*beta exchange could not occur. Adhesion of hepatocytes to terminal galactoside was required by Mg^<2+> but not Ca^<2+>. In contrast, it was suggested that immature hepatocytes could not bind to the same sites because sialic acids were added to the galactosides on hepatocyte surfaces. However, we found it very difficult to isolate this unknown factor from hepatocyte surfaces. During the period of this project, we found that 24 day-hepatocytes after birth were strongly bound to ganglioside GM3. Therefore, we prepared a monoclonal antibody that would inhibit adhesion of hepatocytes to GM3. Then we examined the adhesion of hepatocytes to GM3 by using 6-18 mAb. As a result, it was suggested that 6-18 mAb recognized molecules that transiently founctioned on hepatocyte surfaces, but were expressed in rat of all ages on FACS analysis. From the result of fluorescent immunostaining, 6-18 mAb recognized molecules were localised from the periportal to pericentral regions of the lobule. We examined the binding specificity of the adhesion of hepatocytes to GM3. Hepatocytes recognized the terminal Siaalpha2-3 Gal-of GM3. This recognition was not required for divalent cations. Furthermore, from the result of western blotting analysis following immunoprecipitation, it was suggested that 6-18 mAb recognized molecules indicated 48 KDa. This molecule may be an unknown protein.

  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] Yasuo Kajiyama and Michio Ui: "Switching from α1 to β-subtypes in adrenergic response during primary culture of adult-rat hepatocytes as affected by the cell-to-cell interaction through plasma membranes" Biochem. J. 303. 313-321 (1994)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasuo Kajiyama, Yutaka Sanai, Michio Ui: "Essential role of asialoglycoprotein for plasma-membrane-induced inhibition of the switching from α1to β-subtypes in adrencergic response during primary culture of rat hepatocytes" Biochem. J. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 梶山 泰生、佐内 豊: "分子間相互作用実験法(細胞・糖鎖相互作用実験法)" 羊土社(発行予定),

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yasuo Kajiyama: "Switching from alpha1 to beta-subtypes in adrenergic response during primary culture of adult rat hepatocytes as affected by the cell-to-cell interaction through plasma membranes" Biochemical J.303. 313-321 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yasuo Kajiyama: "Essential role of asialoglycoprotein for plasma-membrane-induced inhibition of the switching from alpha1 to beta-subtypes in adrenergic response during primary culture of rat hepatocytes" Biochemical J.(in press).

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yasuo Kajiyama: Intermolecular interaction Methods in Cell-to Carbohydrate Interactions. Yodosha, (in press),

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1997-03-04  

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