TY - JOUR
T1 - P2-purinoceptors utilize multiple signalling pathways in MDCK-D1 cells
AU - Insel, P. A.
AU - Firestein, B. L.
AU - Xing, M.
AU - Post, S. R.
AU - Jacobson, J. P.
AU - Balboa, M. A.
AU - Hughes, R. J.
N1 - Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 1996
Y1 - 1996
N2 - 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.
AB - 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.
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U2 - 10.1111/j.1474-8673.1996.tb00042.x
DO - 10.1111/j.1474-8673.1996.tb00042.x
M3 - Article
C2 - 9131405
AN - SCOPUS:0030299057
SN - 0144-1795
VL - 16
SP - 311
EP - 314
JO - Journal of Autonomic Pharmacology
JF - Journal of Autonomic Pharmacology
IS - 6
ER -