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Am J Physiol Gastrointest Liver Physiol 280: G795-G804, 2001;
0193-1857/01 $5.00
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Vol. 280, Issue 5, G795-G804, May 2001

P2Y11, a purinergic receptor acting via cAMP, mediates secretion by pancreatic duct epithelial cells

T. D. Nguyen, S. Meichle, U. S. Kim, T. Wong, and M. W. Moody

Department of Medicine, Division of Gastroenterology, University of Washington and Department of Veterans Affairs Puget Sound Health Care System, Seattle, Washington 98108

Pancreatic duct epithelial cells (PDEC) mediate the exocrine secretion of fluid and electrolytes. We previously reported that ATP and UTP interact with P2Y2 receptors on nontransformed canine PDEC to increase intracellular free Ca2+ concentration ([Ca2+]i) and stimulate Ca2+-activated Cl- and K+ channels. We now report that ATP interacts with additional purinergic receptors to increase cAMP and activate Cl- channels. ATP, 2-methylthio-ATP, and ATP-gamma -S stimulated a 4- to 10-fold cAMP increase with EC50 of 10-100 µM. Neither UTP nor adenosine stimulated a cAMP increase, excluding a role for P2Y2 or P1 receptors. Although UTP stimulated an 125I- efflux that was fully inhibited by 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetra(acetoxymethyl) ester (BAPTA-AM), ATP stimulated a partially resistant efflux, suggesting activation of additional Cl- conductances through P2Y2-independent and Ca2+-independent pathways. In Ussing chambers, increased cAMP stimulated a much larger short-circuit current (Isc) increase from basolaterally permeabilized PDEC monolayers than increased [Ca2+]i. Luminal ATP and UTP and serosal UTP stimulated a small Ca2+-type Isc increase, whereas serosal ATP stimulated a large cAMP-type Isc response. Serosal ATP effect was inhibited by P2 receptor blockers and unaffected by BAPTA-AM, supporting ATP activation of Cl- conductances through P2 receptors and a Ca2+-independent pathway. RT-PCR confirmed the presence of P2Y11 receptor mRNA, the only P2Y receptor acting via cAMP.

iodide efflux; adenosine 5'-triphosphate; Ussing chamber


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