Characterization of two distinct chloride channels in cultured dog pancreatic duct epithelial cells. Nguyen, Toan D., Duk-Su Koh[acute]a, Mark W. Moody, Norman R. Fox, Chris E. Savard, Rahul Kuver, Bertil Hille[acute]a, Sum P. Lee. Departments of Medicine and Physiology and Biophysics ([acute]a), University of Washington & Seattle V.A. Medical Center, Seattle, Washington
APStracts 3:0157G, 1996.
Chloride (Cl-) secretion by pancreatic duct epithelial cells (PDEC) regulates cellular bicarbonate secretion, an important component of the exocrine pancreas. In cystic fibrosis, for example, impaired function of the CFTR Cl- channel results in decreased pancreatic secretion and secondary pancreatic insufficiency. Studies of ion transport by PDEC have been hindered by the lack of a practical in -vitro model. We have successfully cultured non-transformed dog PDEC on Vitrogen-coated permeable membranes overlying a feeder layer of myofibroblasts and now report the characterization of Cl- channels in these cells. Cl- conductance, assessed through efflux of 125I- from PDEC, was stimulated by agents acting via cAMP or cytosolic Ca2+. The Cl- conductances activated by cAMP and Ca2+ were distinct since they were differentially inhibited by DIDS, and to a lesser extent, by NPPB and DPC. Patch-clamp studies confirmed the presence of Cl- channels activated by cAMP and Ca2+, with differential inhibition by DIDS. The presence of CFTR Cl- channels in PDEC was confirmed by immunoblotting. These cultured PDEC will be an optimal model for studies of pancreatic duct secretion.

Received 2 October 1995; accepted in final form 6 August 1996.
APS Manuscript Number G423-5.
Article publication pending Am. J. Physiol. (Gastrointest. Liver
Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 29 August 1996