1995 Fiscal Year Final Research Report Summary
A study of the mechanism of intra and inter cellular signal transduction of liver cells.
| Project/Area Number |
04807054
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Gastroenterology
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| Research Institution | Showa University |
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Principal Investigator |
YONEYAMA Keiichiro Showa University School of Medicine, Istructor, 医学部, 講師 (60192431)
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| Project Period (FY) |
1992 – 1994
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| Keywords | intracellular Ca^<2+> / intracellular pH / confocal laser scanning microscopy / gap junction / regenerating rat liver / signal transduction / cell to cell communication |
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| Research Abstract |
(1) Changes in the expression of gap junction protein and their relation to DNA synthesis of hepatocytes in regenerating rat liver were examined immunohistchemically with monoclonal connexin 32 antibody. The results suggest that alteration of liver gap junction induced by galactosamine was not related to cell proliferation, uinlike previously reported alteration after hepatectomy.
(2) The intracellular pH (pHi) of cultured rat hepatocytes stained with BCECF was determined repetitively for minutes confocal laser scanning microscopy. Stable fluorescence under repetitive laser excitation is necessary for monitoring of pHi to study changes of pHi for required time during the reaction. Under the most optimal condition with addition of tis buffers and application of ND filter, the most suitable concentrations of BCECF were determined as 2,4 and 6muM.
(3) In order to analyze the mechanism of intercellular signal transduction, we developed a method to estimate intracellular Ca^<2+>([Ca^<2+>]i) using rat liver tissue slices by confocal laser scanning microscopy (CLSM). It was possible to observe changes in [Ca^<2+>]i in rat cultured liver tissue slices loaded with fluorogenic calcium probe by CLSM.Increase in [Ca^<2+>]i following the induction by [Ca^<2+>]i stimulants was different in the different portious of hepatic acinus and this finding suggests the dofference in intralobular signal transduction in the hepatic acinus. The estimation of [Ca^<2+>]i using rat cultured liver tissue slices by CLSM is useful for the evaluation of intercellular signal transduction in hepatic acinus.
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