P2-purinoceptors utilize multiple signalling pathways in MDCK-D1 cells

P. A. Insel, B. L. Firestein, M. Xing, S. R. Post, J. P. Jacobson, M. A. Balboa, R. J. Hughes

Research output: Contribution to journalArticlepeer-review

Abstract

Madin-Darby canine kidney (MDCK) cells are a widely used model system for the study of epithelial cells. We have utilized a clonal variant, MDCK-D1 to examine signalling by P2-purinoceptors. Several lines of evidence that lead us to conclude that MDCK-D1 cells co-express P(2u) - and P(2y)-purinoceptors and that both subtypes are linked to the release of arachidonic acid and metabolites (AA) include: (a) relative potency of nucleotide analogues in promoting AA release; (b) blockade by the antagonist suramin of response to the P(2y)-selective agonist, 2-methylthio ATP (2-MT-ATP), but not to the P(2u)-selective agonist, UTP; and (c) additivity of response to 2-MT-ATP and UTP. AA release is a consequence of activation of phospholipase A2 (PLA2), most likely the 85 kDa cytosolic PLA2. Treatment of MDCK-D1 cells with ATP, but not UTP, increases inositol 1,4,5-trisphosphate formation while both UTP and ATP increase phosphatidylcholine hydrolysis. ATP, UTP, and 2-MT-ATP can also stimulate phospholipase D activity. Purine nucleotides increase cellular cAMP levels in MDCK-D1 cells in a manner that depends, at least in part, on activation of cyclooxygenase, since cAMP generation stimulated by ATP or UTP is inhibited by treatment of cells with indomethacin. Because cyclooxygenase-derived PGE2 can bind to prostaglandin receptors and stimulate synthesis of cAMP, nucleotides may raise cAMP in an autocrine or paracrine fashion. Taken together, these results indicate that MDCK-D1 cells co-express P(2u) and P(2y)-purinoceptors and that these receptors utilize several mechanisms to regulate cell function, including activation of multiple phospholipases and autocrine/paracrine action of products.

Original languageEnglish (US)
Pages (from-to)311-314
Number of pages4
JournalJournal of Autonomic Pharmacology
Volume16
Issue number6
DOIs
StatePublished - 1996
Externally publishedYes

ASJC Scopus subject areas

  • Neuroscience(all)
  • Pharmacology

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