Properties of the luminal membrane of isolated perfused rat pancreatic ducts. Effect of cyclic AMP and blockers of chloride transport.

The aim of the present study was to investigate by what transport mechanism does HCO-3 cross the luminal membrane of pancreatic duct cells, and how do the cells respond to stimulation with dibutyryl cyclic AMP (db-cAMP). For this purpose a newly developed preparation of isolated and perfused intra- and interlobular ducts of rat pancreas was used. Responses of the epithelium to inhibitors and agonists were monitored by electrophysiological techniques. Addition of HCO-3/CO2 to the bath side of nonstimulated ducts depolarized the PD across the basolateral membrane (PDbl) by about 9 mV, as also observed in a previous study [21]. This HCO-3 effect was abolished by Cl- channel blockers or SITS infused into the lumen of the duct: i.e. 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB, 10(-5) M) hyperpolarized PDbl by 8.2 +/- 1.6 mV (n = 13); 3',5-dichlorodiphenylamine-2-carboxylic acid (DCl-DPC, 10(-5) M) hyperpolarized PDbl by 10.3 +/- 1.7 mV (n = 10); and SITS hyperpolarized PDbl by 7.8 +/- 0.9 mV (n = 4). Stimulation of the ducts with db-cAMP in the presence of bath HCO-3/CO2 resulted in depolarization of PDbl, the ductal lumen became more negative and the fractional resistance of the luminal membrane decreased. Together with forskolin (10(-6) M), db-cAMP (10(-4) M) caused a fast depolarization of PDbl by 33.8 +/- 2.5 mV (n = 6). When db-cAMP (5 x 10(-4) M) was given alone in the presence of bath HCO-3/CO2, PDbl depolarized by 25.3 +/- 4.2 mV (n = 10).(ABSTRACT TRUNCATED AT 250 WORDS)