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Role of cytokeratin intermediate filaments in transhepatic transport and canalicular secretion.The cytoplasmic distribution of ubiquitin was diffuse and particulate.
The effects of nickel persisted 24 hr after its removal from the medium.
These aggregates were associated with intense ubiquitin immunoreactivity.
Only a few attachments of the cytokeratin filaments to the cell cortex remained.
Cytokeratins and ubiquitin were observed using immunofluorescence and immunoelectron microscopy.
F-actin remained attached to the cell cortex in the areas where the cytokeratin filaments had become detached.
Cytokeratin intermediate filaments were selectively dissociated from the cell cortex by nickel treatment.
Further, our study indicates that cytokeratin ubiquitination occurs during collapse and aggregation of the cytokeratin filaments.
The pericanalicular sheath of cytokeratin filaments and the bile canaliculi disappeared and actin was dispersed over the entire cell periphery.
The role of cytokeratin filaments in the function of hepatocytes was investigated using a nickel-treated hepatocyte in vitro model.
Fluorescein diacetate secretion and horseradish peroxidase uptake were almost completely absent in the hepatocytes treated with nickel. Paper-6533.
Hepatocytic function was assessed by visualizing uptake, transhepatic transport and secretion of fluorescein diacetate and horseradish peroxidase into the bile canaliculi. Paper-6533.
The formation of cytokeratin-ubiquitin conjugates during aggregation suggests a role of ubiquitin in the control of cytokeratin organization in hepatocytes in the response to cell stress.
In control primary cultures, most of the bile canaliculi were surrounded by an inner layer of actin filaments and an outer pericanalicular sheath of cytokeratin filaments and microtubules.
It is concluded that cytokeratin intermediate filaments play a critical role in the formation of the bile canaliculus, secretion of fluorescein diacetate and uptake of horseradish peroxidase. Paper-6533.
After treatment with NiCl2 (150 micrograms/ml) for 24 hr, the cytokeratin filaments and desmoplakin became focally detached from the cell cortex and retracted to form an aggregate around the nucleus. Paper-6533.
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