The study of cholestasis using immunoelectron microscopy and rapid freezing deep-etching replica

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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.