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Am J Physiol Renal Physiol 271: F610-F618, 1996;
0363-6127/96 $5.00
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AJP - Renal Physiology, Vol 271, Issue 3 610-F618, Copyright © 1996 by American Physiological Society

ARTICLES

Heterogeneity of P2u- and P2y-purinergic receptor regulation of phospholipases in MDCK cells

B. L. Firestein, M. Xing, R. J. Hughes, C. U. Corvera and P. A. Insel
Department of Pharmacology, School of Medicine, University of California at San Diego, La Jolla 92093-0636, USA.

We have characterized the signaling pathways of purinergic receptors present on the renal epithelial cell line, Madin-Darby canine kidney (MDCK, D1 subclone). Several lines of evidence are consistent with the conclusion that coexisting P2u and P2y receptors release arachidonic acid and metabolites (AA) from MDCK-D1 cells: 1) relative potencies of nucleotide analogues, 2) blockade of P2y agonist- but not P2u agonist-mediated release by suramin, and 3) additivity by 2-methylthio-ATP and UTP. Differences exist between the signaling pathways of the two receptors: pertussis toxin treatment partially inhibits P2u- but not P2y-mediated AA release, and P2y (but not P2u) receptors appear to stimulate D-myo-inositol 1,4,5-trisphosphate production. P2u-receptor occupancy results in both homologous and heterologous desensitization; P2y-receptor occupancy elicits only homologous desensitization. Both receptors stimulate phosphatidylcholine hydrolysis via phospholipase C activation. However, AA release appears to result from phospholipid deacylation by phospholipase A2 activation, rather than from alternate pathways that may include PLC activation. These results demonstrate for the first time that two subtypes of P2-purinergic receptors, P2u and P2y receptors, coexist on a single renal epithelium cell type and that these two receptor subtypes can promote AA release, probably via activation of PLA2.


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