Rearrangement of hepatocellular F-actin precedes the formation of rosette-like structures in parenchyma of cholestatic rat liver.

Abstract

The involvement of hepatocytes in proliferation of bile ductule-like structures during cholestasis remains controversial. The present study was an attempt to address the issue of whether hepatocytes transform into ductular epithelial cells in response to cholestasis and, if so, which mechanisms are involved. Cholestasis was induced by common bile duct-ligation (CBDL) in rat liver for 2, 7, and 14 days. Immunofluorescence microscopy of cytokeratin 19 (CK19) was performed to assess the proliferation of bile ductules. Hepatocellular filamentous actin (F-actin) was studied using fluorescence microscopy of 7-nitrobenzene-2-oxa-1,3-diazole phallacidin and electron microscopy. Double labeling of F-actin and laminin was performed to reveal the relationship between rearrangement of F-actin and deposition of the extracellular matrix protein. The results showed that the localization of F-actin in hepatocytes underwent considerable changes after CBDL, from an even distribution at the entire plasma membrane in control liver to a more concentrated localization at one domain of the plasma membrane. This was followed by formation of rosette-like structures in pericentral and midzonal areas of the parenchyma. CK19 was expressed, as in the control liver, in the epithelial cells of proliferated bile ductules in enlarged portal tracts but not in rosette-like structures of CBDL livers. Furthermore, CBDL induced increasing amounts of laminin in the basal lamina of bile ducts and in connective tissue of portal tracts. In parenchyma, the newly deposited laminin was in close association with the rosette-like structures. It is concluded that the rearrangement of hepatocellular F-actin after CBDL precedes the formation of rosette-like structures. It is speculated that the altered F-actin contracts at one side of hepatocytes leading to tubular structures. Laminin may play an important role in this transformation process.