Role of ADP receptor P2Y(12) in platelet adhesion and thrombus formation in flowing blood.
ADP plays a central role in regulating platelet function. It induces platelet aggregation via the activation of 2 major ADP receptors, P2Y(1) and P2Y(12). We have investigated the role of P2Y(12) in platelet adhesion and thrombus formation under physiological flow by using blood from a patient with a defect in the gene encoding P2Y(12). Anticoagulated blood from the patient and from healthy volunteers was perfused over collagen-coated coverslips. The patient's thrombi were smaller and consisted of spread platelets overlying platelets that were not spread, whereas control thrombi were large and densely packed. Identical platelet surface coverage, aggregate size, and morphology were found when a P2Y(12) antagonist, N(6)-(2-methylthioethyl)-2-(3,3,3-trifluoropropylthio)-beta,gamma-dichloromethylene ATP (also known as AR-C69931 MX), was added to control blood. The addition of a P2Y(1) antagonist (adenosine-3',5'-diphospate) to control blood resulted in small, but normally structured, thrombi. Thus, the ADP-P2Y(12) interaction is essential for normal thrombus buildup on collagen. The patient's blood also showed reduced platelet adhesion on fibrinogen, which was not due to changes in morphology. Comparable results were found by using control blood with AR-C69931 MX and also with adenosine-3',5'-diphospate. This suggested that P2Y(12) and P2Y(1) were both involved in platelet adhesion on immobilized fibrinogen, thereby revealing it as ADP dependent. This was confirmed by complete inhibition on the addition of creatine phosphate/creatine phosphokinase.
Remijn
Jasper A
JA
Thrombosis and Haemostasis Laboratory, Department of Haematology, University Medical Center Utrecht, the Netherlands. J.A.Remijn@lab.azu.nl
Wu
Ya-Ping
YP
Jeninga
Ellen H
EH
IJsseldijk
Martin J W
MJ
van Willigen
Gijsbert
G
de Groot
Philip G
PG
Sixma
Jan J
JJ
Nurden
Alan T
AT
Nurden
Paquita
P
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000249:Adenosine Monophosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000368:Aged; D000704:Analysis of Variance; D001792:Blood Platelets / Q000187:drug effects; D006801:Humans; D008297:Male; D008565:Membrane Proteins; D010973:Platelet Adhesiveness / Q000187:drug effects / Q000502:physiology*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D018048:Receptors, Purinergic P2 / Q000235:genetics / Q000502:physiology*; D058925:Receptors, Purinergic P2Y12; D013927:Thrombosis / Q000209:etiology* / Q000473:pathology
D008565:Membrane Proteins; C548576:P2RY12 protein, human; D010975:Platelet Aggregation Inhibitors; D018048:Receptors, Purinergic P2; D058925:Receptors, Purinergic P2Y12; D000249:Adenosine Monophosphate; D000244:Adenosine Diphosphate; C117446:cangrelor; C010840:adenosine 3'-phosphate-5'-phosphate