The role of the hepatocyte cytokeratin network in bile formation and resistance to bile acid challenge and cholestasis in mice.
The intermediate filament cytoskeleton of hepatocytes is composed of keratin (K) 8 and K18 and has important mechanical and nonmechanical functions. However, the potential role of the K8/K18 network for proper membrane targeting of hepatocellular adenosine triphosphate-binding cassette transporters and bile formation is unknown. We therefore designed a comparative study in K8 and K18 knockout mice and respective wild-type controls to test the hypothesis that intermediate filaments of hepatocytes play a role in normal bile formation. In addition, we challenged mice either with a 1% cholic acid-supplemented diet or a diet containing the porphyrinogenic xenobiotic 3,5-diethoxycarbonyl-1,4-dihydrocollidine to determine the effect of K8/K18 loss on bile flow/composition and liver injury under different physiological and toxic stress stimuli. Protein expression levels and membrane localization of various transporters and anion exchangers were compared using western blotting and immunofluorescence microscopy, respectively, and bile flow and composition were determined under various experimental conditions. Our results demonstrate that loss of the intermediate filament network had no significant effect on bile formation and composition, as well as expression levels and membrane targeting of key hepatobiliary transporters under baseline and stress conditions. However, loss of K8 significantly increased liver injury in response to toxic stress.


CONCLUSIONS
The intermediate filament network of hepatocytes is not specifically required for proper bile formation in mice.
Fickert
Peter
P
Laboratory of Experimental and Molecular Hepatology, Division of Gastroenterology and Hepatology, Medical University of Graz, Graz, Austria.
Fuchsbichler
Andrea
A
Wagner
Martin
M
Silbert
Dagmar
D
Zatloukal
Kurt
K
Denk
Helmut
H
Trauner
Michael
M
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000818:Animals; D001646:Bile / Q000378:metabolism*; D001647:Bile Acids and Salts / Q000097:blood; D002779:Cholestasis / Q000235:genetics / Q000503:physiopathology*; D019826:Cholic Acid / Q000494:pharmacology; D022781:Hepatocytes / Q000378:metabolism*; D007382:Intermediate Filaments / Q000187:drug effects / Q000378:metabolism*; D053538:Keratin-18 / Q000378:metabolism; D053533:Keratin-8 / Q000378:metabolism; D007633:Keratins / Q000378:metabolism*; D008099:Liver / Q000201:enzymology / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout
D018528:ATP-Binding Cassette Transporters; D001647:Bile Acids and Salts; D053538:Keratin-18; D053533:Keratin-8; D007633:Keratins; D019826:Cholic Acid