Novel vasocontractile role of the P2Y14receptor: characterization of its signalling in porcine isolated pancreatic arteries
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 P2Y1...
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| Format: | Article |
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Wiley
2014
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| Online Access: | https://eprints.nottingham.ac.uk/34818/ |
| _version_ | 1848794942352130048 |
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| author | Alsaqati, M. Latif, M.L. Chan, S.L.F. Ralevic, V. |
| author_facet | Alsaqati, M. Latif, M.L. Chan, S.L.F. Ralevic, V. |
| author_sort | Alsaqati, M. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T19:24:12Z |
| format | Article |
| id | nottingham-34818 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:24:12Z |
| publishDate | 2014 |
| publisher | Wiley |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-348182020-05-04T16:43:24Z https://eprints.nottingham.ac.uk/34818/ Novel vasocontractile role of the P2Y14receptor: characterization of its signalling in porcine isolated pancreatic arteries Alsaqati, M. Latif, M.L. Chan, S.L.F. Ralevic, V. 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. Wiley 2014-02-04 Article PeerReviewed Alsaqati, M., Latif, M.L., 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 UDP-glucose; UDP;MRS2690; P2Y14 receptor; vasoconstriction; endothelium http://onlinelibrary.wiley.com/doi/10.1111/bph.12473/abstract;jsessionid=804DA88F606FEAA0EC72BFAE4C608687.f01t02 doi:10.1111/bph.12473 doi:10.1111/bph.12473 |
| spellingShingle | UDP-glucose; UDP;MRS2690; P2Y14 receptor; vasoconstriction; endothelium Alsaqati, M. Latif, M.L. Chan, S.L.F. Ralevic, V. Novel vasocontractile role of the P2Y14receptor: characterization of its signalling in porcine isolated pancreatic arteries |
| title | Novel vasocontractile role of the P2Y14receptor: characterization of its signalling in porcine isolated pancreatic arteries |
| title_full | Novel vasocontractile role of the P2Y14receptor: characterization of its signalling in porcine isolated pancreatic arteries |
| title_fullStr | Novel vasocontractile role of the P2Y14receptor: characterization of its signalling in porcine isolated pancreatic arteries |
| title_full_unstemmed | Novel vasocontractile role of the P2Y14receptor: characterization of its signalling in porcine isolated pancreatic arteries |
| title_short | Novel vasocontractile role of the P2Y14receptor: characterization of its signalling in porcine isolated pancreatic arteries |
| title_sort | novel vasocontractile role of the p2y14receptor: characterization of its signalling in porcine isolated pancreatic arteries |
| topic | UDP-glucose; UDP;MRS2690; P2Y14 receptor; vasoconstriction; endothelium |
| url | https://eprints.nottingham.ac.uk/34818/ https://eprints.nottingham.ac.uk/34818/ https://eprints.nottingham.ac.uk/34818/ |