title: Detection of P2Y(14) protein in platelets and investigation of the role of P2Y(14) in platelet function in comparison with the EP(3) receptor.
abstract: mRNA encoding the recently discovered P2Y(14) receptor has been reported in platelets, but the presence of P2Y(14) receptor protein and its functionality have not been studied. If P2Y(14) is expressed along with P2Y(1) and P2Y(12) receptors it may have a role in haemostasis. It was the objective of this study to investigate the presence of the P2Y(14) receptor in platelets and its role in platelet function. The effects of the agonist UDP-glucose were compared with those of sulprostone, a selective EP(3) receptor agonist. Expression of P2Y(14) receptor was investigated by immunoblotting and confocal microscopy. Platelet aggregation in platelet-rich plasma (PRP) and whole blood was measured using light absorbance and platelet counting. VASP phosphorylation was investigated using flow cytometry. Immunoblotting provided evidence for P2Y(14) receptor protein and microscopy confirmed its presence on platelets. Despite this, UDP-glucose (up to 100 muM) did not induce platelet aggregation in either PRP or whole blood, and did not potentiate aggregation induced by other agonists. P2Y(14) did not substitute for P2Y(12) in experiments using the P2Y(12) antagonist AR-C69931. No effect of UDP-glucose was seen on adenylate cyclase activity as measured by VASP phosphorylation. In contrast, sulprostone acting via the EP(3) receptor promoted platelet aggregation with effects on adenylate cyclase activity. EP(3) also partially substituted for P2Y(12) receptor. We have demonstrated the presence of P2Y(14) receptor protein in platelets, but no contribution of this receptor to several measures of platelet function has been observed. Further studies are necessary to determine whether the P2Y(14) receptor in platelets has any functionality.
authors.affiliation: Cardiovascular Medicine, Queen's Medical Centre, Nottingham, UK.
authors.forename: Natalia
authors.initials: N
authors.lastname: Dovlatova
authors.forename: Yanushi D
authors.initials: YD
authors.lastname: Wijeyeratne
authors.forename: Susan C
authors.initials: SC
authors.lastname: Fox
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Manolopoulos
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Johnson
authors.forename: Ann E
authors.initials: AE
authors.lastname: White
authors.forename: M Liaque
authors.initials: ML
authors.lastname: Latif
authors.forename: Vera
authors.initials: V
authors.lastname: Ralevic
authors.forename: Stanley
authors.initials: S
authors.lastname: Heptinstall
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology; D000249:Adenosine Monophosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000378:metabolism; D015232:Dinoprostone / Q000031:analogs & derivatives / Q000494:pharmacology; D006801:Humans; D015151:Immunoblotting; D008840:Microfilament Proteins / Q000378:metabolism; D018613:Microscopy, Confocal; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D010974:Platelet Aggregation / Q000187:drug effects / Q000502:physiology*; D010976:Platelet Count; D058910:Purinergic P2 Receptor Agonists; D051381:Rats; D018078:Receptors, Prostaglandin E / Q000819:agonists / Q000378:metabolism*; D058308:Receptors, Prostaglandin E, EP3 Subtype; D018048:Receptors, Purinergic P2 / Q000302:isolation & purification / Q000378:metabolism*; D014532:Uridine Diphosphate Glucose / Q000494:pharmacology; D000098662:Vasodilator-Stimulated Phosphoprotein
chemical_list: D015815:Cell Adhesion Molecules; D008840:Microfilament Proteins; C518528:P2Y14 receptor, human; C545647:PTGER3 protein, human; D010750:Phosphoproteins; D058910:Purinergic P2 Receptor Agonists; D018078:Receptors, Prostaglandin E; D058308:Receptors, Prostaglandin E, EP3 Subtype; D018048:Receptors, Purinergic P2; D000098662:Vasodilator-Stimulated Phosphoprotein; D000249:Adenosine Monophosphate; C016767:sulprostone; D000244:Adenosine Diphosphate; C117446:cangrelor; D015232:Dinoprostone; D014532:Uridine Diphosphate Glucose