1990 Fiscal Year Final Research Report Summary
Clarification of Biological Meaning of Gap-Junctional Intercellular Communication by Using Monoclonal Antibodies
| Project/Area Number |
62440022
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
General physiology
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| Research Institution | Hiroshima University |
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Principal Investigator |
KANNO Yoshinobu Hiroshima University school of Dentistry, Professor, 歯学部, 教授 (00034158)
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| Co-Investigator(Kenkyū-buntansha) |
HIRONO Chikara Hiroshima University school of Dentistry, Research Associate, 歯学部, 助手 (10199135)
SHIBA Yoshiki Hiroshima University school of Dentistry, Associate Professor, 歯学部, 助教授 (90110452)
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| Project Period (FY) |
1987 – 1990
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| Keywords | Gap junctions / Intercellular communication / Monoclonal antibody / Rat liver / Rat salivary gland / Cultured cells / Cell proliferation / Secretion |
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| Research Abstract |
In this project, we investigated the biological meaning of gap-junctional intercellular communication, by using monoclonal antibodies against gap junction proteins. Gap junction protein (27KDa) was purified from sodium hydrochoride-insoluble fraction of rat liver homogenate, and a monoclonal antibody against purified gap junction protein was obtained according to the conventional procedure. Monoclonal antibody (14-84Al) recognized 27 KDa gap junction protein in the crude membrane fraction from rat liver, and stained the interecellular membrane of hepatocytes as spotted appearance. To clarify the biological meaning of gap junctional communication, its relationship to the cell proliferation and the secretory activity was investigated. The gap junction proteins in the isolated hepatocytes rapidly disappeared after culturing in vitro. These results suggest that gap-junctional intercellular communication is not required in the proliferating hepatocytes. The gap-junctional communication in cultured 3T3-L1 cells also exerted suppressive action on the proliferation.
The gap-junctional communication in the salivary gland acinar cells was related to the activity of the acinar cells to secrete the salivary juice.
The control of the expression of gap junction protein was also investigated by using monoclonal antibodies. Cultured cells derived from the liver was not stained with 14-84Al monoclonal antibody, but these cell lines exhibited well-developed dye-coupling, suggesting the modulation of the expression of gap junction proteins by in vitro cultures. The gap junctions of acinar cells in rat salivary glands were composed of 27 KDa gap junction proteins, and required the innervation of autonomic nerve system for its maintenance. Further studies using monoclonal antibodies against an other gap junctional proteins are necessary to clarify the control mechanism of the expression of gap junction proteins.
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