The study of cholestasis using immunoelectron microscopy and rapid freezing deep-etching replica
| Project/Area Number |
62480198
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Gastroenterology
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
OKANOUE Takeshi Kyoto Prefectural University of Medicine, Department of Medicine, Associate Professor, 医学部, 助教授 (20150568)
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| Co-Investigator(Kenkyū-buntansha) |
MORIOKA Hiroyuki Kyoto Prefectural University of Medicine, Department of Medicine, Assistant Prof, 医学部, 助手 (50079951)
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| Project Period (FY) |
1987 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 1989: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1988: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1987: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Keywords | Intrahepatic cholestasis / Cytoskeleton of hepatocyte / Intermediate filament / Microfilament / Pericanalicular sheath / Desmosome / Immunoelectron microscopy / 肝内胆汁うつ滞 / 肝細胞中間径フィラメント / 肝細胞マイクロフィラメント / デスモプラキン / 胆細胞細胞骨格 / 免疫螢光法 / 走査電顕 / 急速凍結ディープエッチングレプリカ法 |
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| Research Abstract |
We investigated the cytoskeletal changes of hepatocytes in various kinds of cholestatic rat models using immunofluorescence, transmission electron microscope(TEM) and immunoelectron microscope(IEM). The rats treated with phalloidin, cytokalasin B and cholestatic factor were used as the model of in- trahepatic cholestasis and the rats as the model of obstructive jaundice.
Pericanalicular sheath(PS) was clearly demonstrated by TEM in the liver tissues treated by the method of Katsuma et al. Almost all filaments made up PS were identified as intermediate filaments(IFs) by IEM.
In the phalloidin treated and bile duct ligated rats, microfilaments(MFS) and IFs increased in amounts and PS was dilated and thickened. The focal disruption of the PS was observed in the bile duct ligated rats, which disappeared after recanalization of the bile ducts. These results suggest that IFs increase in amounts according to the increase of MFs in cholestasis.
We also investigated the immunological hetetogeneity of IFs in hepatocytes using a few kinds of monoclonal anti cytokeratin(CK) antibodies including No.8 and No. 18(Moll's classification) and the change of IFs of hepatocytes was noted in the hyperplastic nodules in the experimental model of hepatocacinogenesis. Immunofluorescence and immunoelectron microscopy demonstrated that the staining pattern of cytokeratin polypeptides changed in the process of hepatocarcinogenesis, which might be useful in the early detection of hepatocellular carcinoma.
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Report
(4 results)
Research Products
(23 results)
