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Am J Physiol Renal Physiol 279: F1045-F1052, 2000;
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Vol. 279, Issue 6, F1045-F1052, December 2000

P2Y2 receptor of MDCK cells: cloning, expression, and cell-specific signaling

Alexander C. Zambon, Richard J. Hughes, J. Gary Meszaros, J. Julie Wu, Brian Torres, Laurence L. Brunton, and Paul A. Insel

Departments of Pharmacology and Medicine, Biomedical Sciences Graduate Program, University of California at San Diego, La Jolla, California 92093-0636

Madin-Darby canine kidney (MDCK)-D1 cells, a canine renal epithelial cell line, co-express at least three different P2Y receptor subtypes: P2Y1, P2Y2, and P2Y11 (24). Stimulation of P2Y receptors in these cells results in the release of arachidonic acid (AA) and metabolites and the elevation of intracellular cAMP. To define in more precise terms the signaling contributed by the MDCK-D1 P2Y2 (cP2Y2) receptor, we have cloned and heterologously expressed it in CF2Th (canine thymocyte) cells, a P2Y2-null cell. Analysis by RT-PCR indicated that canine P2Y2 receptors are expressed in skeletal muscle, spleen, kidney, lung, and liver. When expressed in CF2Th cells, cP2Y2 receptors promoted phospholipase C-mediated phosphatidylinositol (PI) hydrolysis [uridine 5'-triphosphate >=  ATP > adenosine 5'-diphosphate > 2MT-ATP] and mobilization of intracellular Ca2+. In contrast to their actions in MDCK-D1 cells, cP2Y2 receptors did not stimulate formation of cAMP or AA release when expressed in CF2Th cells. The data indicate that cell setting plays an essential role in the ability of P2Y receptors to regulate AA release and cAMP formation. In particular, renal epithelial cells preferentially express components critical for cP2Y2-induced cAMP formation, including the expression of enzymes involved in the generation and metabolism of AA and receptors that respond to PGE2.

Madin-Darby canine kidney; epithelial cells; canine P2Y2 receptors; adenosine 3',5'-cyclic monophosphate; CF2Th thymocytes


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