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Am J Physiol Gastrointest Liver Physiol 279: G132-G138, 2000;
0193-1857/00 $5.00
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Vol. 279, Issue 1, G132-G138, July 2000

Extracellular UTP stimulates electrogenic bicarbonate secretion across CFTR knockout gallbladder epithelium

Lane L. Clarke1, Matthew C. Harline1, Lara R. Gawenis1, Nancy M. Walker1, John T. Turner2, and Gary A. Weisman3

1 Dalton Cardiovascular Research Center and Departments of Veterinary Biomedical Sciences, 2 Pharmacology, and 3 Biochemistry, University of Missouri-Columbia, Columbia, Missouri 65211

The loss of cystic fibrosis transmembrane conductance regulator (CFTR)-mediated transepithelial HCO3- secretion contributes to the pathogenesis of pancreatic and biliary disease in cystic fibrosis (CF) patients. Recent studies have investigated P2Y2 nucleotide receptor agonists, e.g., UTP, as a means to bypass the CFTR defect by stimulating Ca2+-activated Cl- secretion. However, the value of this treatment in facilitating transepithelial HCO3- secretion is unknown. Gallbladder mucosae from CFTR knockout mice were used to isolate the Ca2+-dependent anion conductance during activation of luminal P2Y2 receptors. In Ussing chamber studies, UTP stimulated a transient peak in short-circuit current (Isc) that declined to a stable plateau phase lasting 30-60 min. The plateau Isc after UTP was Cl- independent, HCO3- dependent, insensitive to bumetanide, and blocked by luminal DIDS. In pH stat studies, luminal UTP increased both Isc and serosal-to-mucosal HCO3- flux (Jsright-arrow m) during a 30-min period. Substitution of Cl- with gluconate in the luminal bath to inhibit Cl-/HCO3- exchange did not prevent the increase in Jsright-arrow m and Isc during UTP. In contrast, luminal DIDS completely inhibited UTP-stimulated increases in Jsright-arrow m and Isc. We conclude that P2Y2 receptor activation results in a sustained (30-60 min) increase in electrogenic HCO3- secretion that is mediated via an intracellular Ca2+-dependent anion conductance in CF gallbladder.

cystic fibrosis; biliary system; P2Y2 receptor; nucleotide receptor; purinoceptor; chloride


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