Pharmacological Characterization of Nucleotide P2Y Receptors on Endothelial Cells of the Mouse Aorta

Overview

Journal Br J Pharmacol

Publisher Wiley

Specialty Pharmacology

Date 2005 Jul 6

PMID 15997227

Citations 35

Authors

Pieter-Jan D F Guns

Andras Korda

Herta M Crauwels

Tim Van Assche

Bernard Robaye

Jean-Marie Boeynaems

Hidde Bult

Affiliations

Soon will be listed here.

Abstract

Nucleotides regulate various effects including vascular tone. This study was aimed to characterize P2Y receptors on endothelial cells of the aorta of C57BL6 mice. Five adjacent segments (width 2 mm) of the thoracic aorta were mounted in organ baths to measure isometric force development. Nucleotides evoked complete (adenosine 5' triphosphate (ATP), uridine 5' triphosphate (UTP), uridine 5' diphosphate (UDP); >90%) or partial (adenosine 5' diphosphate (ADP)) relaxation of phenylephrine precontracted thoracic aortic rings of C57BL6 mice. Relaxation was abolished by removal of the endothelium and was strongly suppressed (>90%) by inhibitors of nitric oxide synthesis. The rank order of potency was: UDP approximately UTP approximately ADP>adenosine 5'-[gamma-thio] triphosphate (ATPgammaS)>ATP, with respective pD2 values of 6.31, 6.24, 6.22, 5.82 and 5.40. These results are compatible with the presence of P2Y1 (ADP>ATP), P2Y2 or P2Y4 (ATP and UTP) and P2Y6 (UDP) receptors. P2Y4 receptors were not involved, since P2Y4-deficient mice displayed unaltered responses to ATP and UTP. The purinergic receptor antagonist suramin exerted surmountable antagonism for all agonists. Its apparent pKb for ATP (4.53+/-0.07) was compatible with literature, but the pKb for UTP (5.19+/-0.03) was significantly higher. This discrepancy suggests that UTP activates supplementary non-P2Y2 receptor subtype(s). Further, pyridoxal-phosphate-6-azophenyl-2'-4'-disulphonic acid (PPADS) showed surmountable (UTP, UDP), nonsurmountable (ADP) or no antagonism (ATP). Finally, 2'-deoxy-N6-methyladenosine3',5'-bisphosphate (MRS2179) inhibited ADP-evoked relaxation only. Taken together, these results point to the presence of functional P2Y1 (ADP), P2Y2 (ATP, UTP) and P2Y6 (UDP) receptors on murine aorta endothelial cells. The identity of the receptor(s) mediating the action of UTP is not fully clear and other P2Y subtypes might be involved in UTP-evoked vasodilatation.

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References

Talukder M, Morrison R, Mustafa S . Comparison of the vascular effects of adenosine in isolated mouse heart and aorta. Am J Physiol Heart Circ Physiol. 2001; 282(1):H49-57. DOI: 10.1152/ajpheart.2002.282.1.H49. View

van der Giet M, Giebing G, Tolle M, Schmidt S . The Role of P2Y Receptors in the Control of Blood Pressure. Drug News Perspect. 2003; 15(10):640-646. DOI: 10.1358/dnp.2002.15.10.740237. View

John K, Barakat A . Modulation of ATP/ADP concentration at the endothelial surface by shear stress: effect of flow-induced ATP release. Ann Biomed Eng. 2001; 29(9):740-51. DOI: 10.1114/1.1397792. View

Palmer R, Boyer J, Schachter J, Nicholas R, Harden T . Agonist action of adenosine triphosphates at the human P2Y1 receptor. Mol Pharmacol. 1998; 54(6):1118-23. View

Yamamoto K, Korenaga R, Kamiya A, Ando J . Fluid shear stress activates Ca(2+) influx into human endothelial cells via P2X4 purinoceptors. Circ Res. 2000; 87(5):385-91. DOI: 10.1161/01.res.87.5.385. View

Vial C, Evans R . P2X(1) receptor-deficient mice establish the native P2X receptor and a P2Y6-like receptor in arteries. Mol Pharmacol. 2002; 62(6):1438-45. DOI: 10.1124/mol.62.6.1438. View

Hansmann G, Bultmann R, Tuluc F, Starke K . Characterization by antagonists of P2-receptors mediating endothelium-dependent relaxation in the rat aorta. Naunyn Schmiedebergs Arch Pharmacol. 1997; 356(5):641-52. DOI: 10.1007/pl00005101. View

Pouillon V, Boeynaems J, Robaye B . Molecular cloning and characterization of the mouse P2Y4 nucleotide receptor. Eur J Pharmacol. 2001; 416(3):197-202. DOI: 10.1016/s0014-2999(01)00875-5. View

Ralevic V, Burnstock G . Receptors for purines and pyrimidines. Pharmacol Rev. 1998; 50(3):413-92. View

10.

Kunapuli S, Daniel J . P2 receptor subtypes in the cardiovascular system. Biochem J. 1998; 336 ( Pt 3):513-23. PMC: 1219898. DOI: 10.1042/bj3360513. View

11.

Lazarowski E, Rochelle L, ONeal W, Ribeiro C, Grubb B, Zhang V . Cloning and functional characterization of two murine uridine nucleotide receptors reveal a potential target for correcting ion transport deficiency in cystic fibrosis gallbladder. J Pharmacol Exp Ther. 2001; 297(1):43-9. View

12.

Harden T, Lazarowski E, Boucher R . Release, metabolism and interconversion of adenine and uridine nucleotides: implications for G protein-coupled P2 receptor agonist selectivity. Trends Pharmacol Sci. 1997; 18(2):43-6. View

13.

Kumari R, Goh G, Ng L, Boarder M . ATP and UTP responses of cultured rat aortic smooth muscle cells revisited: dominance of P2Y2 receptors. Br J Pharmacol. 2003; 140(7):1169-76. PMC: 1574131. DOI: 10.1038/sj.bjp.0705526. View

14.

Beny J . Characterization of purine receptors in mouse thoracic aorta. J Cardiovasc Pharmacol. 2004; 44(2):171-7. DOI: 10.1097/00005344-200408000-00005. View

15.

Crauwels H, Van Hove C, Holvoet P, Herman A, Bult H . Plaque-associated endothelial dysfunction in apolipoprotein E-deficient mice on a regular diet. Effect of human apolipoprotein AI. Cardiovasc Res. 2003; 59(1):189-99. DOI: 10.1016/s0008-6363(03)00353-5. View

16.

Boarder M, Hourani S . The regulation of vascular function by P2 receptors: multiple sites and multiple receptors. Trends Pharmacol Sci. 1998; 19(3):99-107. DOI: 10.1016/s0165-6147(98)01170-5. View

17.

Crauwels H, Van Hove C, Herman A, Bult H . Heterogeneity in relaxation mechanisms in the carotid and the femoral artery of the mouse. Eur J Pharmacol. 2000; 404(3):341-51. DOI: 10.1016/s0014-2999(00)00619-1. View

18.

Bowler J, Bailey R, North R, Surprenant A . P2X4, P2Y1 and P2Y2 receptors on rat alveolar macrophages. Br J Pharmacol. 2003; 140(3):567-75. PMC: 1574050. DOI: 10.1038/sj.bjp.0705459. View

19.

Rubino A, Ziabary L, Burnstock G . Regulation of vascular tone by UTP and UDP in isolated rat intrapulmonary arteries. Eur J Pharmacol. 1999; 370(2):139-43. DOI: 10.1016/s0014-2999(99)00150-8. View

20.

Communi D, Parmentier M, Boeynaems J . Cloning, functional expression and tissue distribution of the human P2Y6 receptor. Biochem Biophys Res Commun. 1996; 222(2):303-8. DOI: 10.1006/bbrc.1996.0739. View