Novel vasocontractile role of the P2Y14receptor: characterization of its signalling in porcine isolated pancreatic arteries

Alsaqati, M. and Latif, M.L. and Chan, S.L.F. and Ralevic, V. (2014) Novel vasocontractile role of the P2Y14receptor: characterization of its signalling in porcine isolated pancreatic arteries. British Journal of Pharmacology, 171 (3). pp. 701-713. ISSN 0007-1188

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Abstract

Background and Purpose: The P2Y14 receptor is the newest member of the P2Y receptor family; it is Gi/o protein-coupled and is activated by UDP and selectively by UDP-glucose and MRS2690 (2-thiouridine-5′-diphosphoglucose) (7–10-fold more potent than UDP-glucose). This study investigated whether P2Y14 receptors were functionally expressed in porcine isolated pancreatic arteries.

Experimental Approach: Pancreatic arteries were prepared for isometric tension recording and UDP-glucose, UDP and MRS2690 were applied cumulatively after preconstriction with U46619, a TxA2 mimetic. Levels of phosphorylated myosin light chain 2 (MLC2) were assessed with Western blotting. cAMP concentrations were assessed using a competitive enzyme immunoassay kit.

Key Results: Concentration-dependent contractions with a rank order of potency of MRS2690 (10-fold) > UDP-glucose ≥ UDP were recorded. These contractions were reduced by PPTN {4-[4-(piperidin-4-yl)phenyl]-7-[4-(trifluoromethyl)phenyl]-2-naphthoic acid}, a selective antagonist of P2Y14 receptors, which did not affect responses to UTP. Contraction to UDP-glucose was not affected by MRS2578, a P2Y6 receptor selective antagonist. Raising cAMP levels and forskolin, in the presence of U46619, enhanced contractions to UDP-glucose. In addition, UDP-glucose and MRS2690 inhibited forskolin-stimulated cAMP levels. Removal of the endothelium and inhibition of endothelium-derived contractile agents (TxA2, PGF2α and endothelin-1) inhibited contractions to UDP glucose. Y-27632, nifedipine and thapsigargin also reduced contractions to the agonists. UDP-glucose and MRS2690 increased MLC2 phosphorylation, which was blocked by PPTN.

Conclusions and Implications: P2Y14 receptors play a novel vasocontractile role in porcine pancreatic arteries, mediating contraction via cAMP-dependent mechanisms, elevation of intracellular Ca2+ levels, activation of RhoA/ROCK signalling and MLC2, along with release of TxA2, PGF2α and endothelin-1.

Item Type: Article
Additional Information: This is the peer reviewed version of the following article: Alsaqati, M., Latif, M. L., Chan, S. L. F. and Ralevic, V. (2014), Novel vasocontractile role of the P2Y14 receptor: characterization of its signalling in porcine isolated pancreatic arteries. British Journal of Pharmacology, 171: 701–713. doi: 10.1111/bph.12473, which has been published in final form at http://onlinelibrary.wiley.com/doi/10.1111/bph.12473. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
Keywords: UDP-glucose; UDP;MRS2690; P2Y14 receptor; vasoconstriction; endothelium
Schools/Departments: University of Nottingham UK Campus > Faculty of Medicine and Health Sciences > School of Life Sciences
Identification Number: https://doi.org/10.1111/bph.12473
Depositing User: Eprints, Support
Date Deposited: 11 Jul 2016 12:51
Last Modified: 22 Sep 2016 20:59
URI: http://eprints.nottingham.ac.uk/id/eprint/34818

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