Molecular and functional heterogeneity of cholangiocytes from rat liver after bile duct ligation. Alpini, Gianfranco, Charles Ulrich, Stuart Roberts, John O. Phillips, Yoshiyuki Ueno, Prasad V. Podila, Oscar Colegio, Gene D. Lesage, Laurence J. Miller, Nicholas F. Larusso. Center for Basic Research in Digestive Diseases, Division of Gastroenterology and Internal Medicine, and Department of Biochemistry and Molecular Biology, Mayo Medical School, Clinic, and Foundation, Rochester, MN, 55905
APStracts 3:0178G, 1996.
Cholangiocytes, the epithelial cells that line intrahepatic bile ducts, participate in bile secretion via basal and agonist-stimulated absorption and secretion of solutes and water. Based on subtle structural differences between cholangiocytes lining small versus large bile ducts, as well as known phenotypic variations among transporting epithelia in other organs (e.g., renal tubules and small intestine), we demonstrated that cholangiocytes are functionally heterogeneous along the intrahepatic biliary tree of normal rats. In studies reported here, we confirm and extend the concept of functional heterogeneity of cholangiocytes by employing the bile duct ligated rat, an important model of cholestasis associated with selective cholangiocyte proliferation, commonly used to evaluate bile duct secretory processes. Using novel isolation and separatory techniques, we prepared subpopulations of essentially pure small, medium, and large cholangiocytes from bile duct ligated rats and compared them with regard to gene expression and basal or agonist -responsive transport activities. While similar levels of transcripts for [gamma]-glutamyltranspeptidase and cytokeratin 19, two cholangiocyte-specific proteins, and glyceraldehyde-3-phosphate dehydrogenase, a house-keeping gene, were present in all three subpopulations, genes for several proteins involved in solute transport [Cl-/HCO3- exchanger, cystic fibrosis transmembrane regulator (CFTR) and secretin receptor (SR)] were expressed only in medium and large cholangiocytes. Consistent with these findings, secretin increased intracellular levels of cAMP and 36Cl- efflux rates in medium and large but not small cholangiocytes. Also, forskolin/cpt-cAMP stimulated 36Cl- efflux rates only in medium and large cholangiocytes, consistent with selective functional expression of CFTR in these subpopulations. These results support the molecular and functional heterogeneity of cholangiocytes within the intrahepatic biliary ductal system and are consistent with the notion that hormone-regulated transport of solutes after bile duct ligation occurs principally in medium and large cholangiocytes in a fashion similar to that observed in normal rat liver.

Received 1 March 1996; accepted in final form 30 August 1996.
APS Manuscript Number G81-6.
Article publication pending Am. J. Physiol. (Gastrointest. Liver
Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 19 September 1996