title: P2Y(2) receptor of MDCK cells: cloning, expression, and cell-specific signaling.
abstract: Madin-Darby canine kidney (MDCK)-D1 cells, a canine renal epithelial cell line, co-express at least three different P2Y receptor subtypes: P2Y(1), P2Y(2), and P2Y(11) (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 P2Y(2) (cP2Y(2)) receptor, we have cloned and heterologously expressed it in CF2Th (canine thymocyte) cells, a P2Y(2)-null cell. Analysis by RT-PCR indicated that canine P2Y(2) receptors are expressed in skeletal muscle, spleen, kidney, lung, and liver. When expressed in CF2Th cells, cP2Y(2) receptors promoted phospholipase C-mediated phosphatidylinositol (PI) hydrolysis [uridine 5'-triphosphate > or = ATP > adenosine 5'-diphosphate > 2MT-ATP] and mobilization of intracellular Ca(2+). In contrast to their actions in MDCK-D1 cells, cP2Y(2) 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 cP2Y(2)-induced cAMP formation, including the expression of enzymes involved in the generation and metabolism of AA and receptors that respond to PGE(2).
authors.affiliation: Department of Pharmacology, Biomedical Sciences Graduate Program, University of California at San Diego, La Jolla, California 92093-0636, USA.
authors.forename: A C
authors.initials: AC
authors.lastname: Zambon
authors.forename: R J
authors.initials: RJ
authors.lastname: Hughes
authors.forename: J G
authors.initials: JG
authors.lastname: Meszaros
authors.forename: J J
authors.initials: JJ
authors.lastname: Wu
authors.forename: B
authors.initials: B
authors.lastname: Torres
authors.forename: L L
authors.initials: LL
authors.lastname: Brunton
authors.forename: P A
authors.initials: PA
authors.lastname: Insel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000096:biosynthesis; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D010716:Phosphatidylinositols / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D018048:Receptors, Purinergic P2 / Q000096:biosynthesis / Q000378:metabolism*; D058473:Receptors, Purinergic P2Y2; D012190:Retroviridae / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology*; D013045:Species Specificity; D013601:T-Lymphocytes / Q000378:metabolism
chemical_list: C546385:P2RY2 protein, human; C546386:P2ry2 protein, mouse; D010716:Phosphatidylinositols; D018048:Receptors, Purinergic P2; D058473:Receptors, Purinergic P2Y2; D016718:Arachidonic Acid; D012313:RNA; D004247:DNA; D000242:Cyclic AMP; D002118:Calcium