The balance of concurrent aggregation and deaggregation processes in platelets is linked to differential occupancy of ADP receptor subtypes.
Deaggregation, the partial reversal of the initial aggregation of platelets is observed following low, but not higher, micromolar ADP concentrations. This study tested the hypothesis that deaggregation results from a balance between concurrent, opposing, aggregation and deaggregation processes which are ADP (adenosine 5'-diphosphate) receptor occupancy-dependent. Aggregation of human platelet-rich plasma (PRP) prepared in r-hirudin was assayed in a 96-well plate reader over 20 min by measurement of the optical density (OD) at 580 nm. Aggregation and the time to reach peak aggregation were directly proportional to ADP receptor occupancy. The magnitude and time course of the response to ADP were comparable to those previously reported with standard aggregometry. The rate constant of platelet deaggregation, as assessed by a four-compartment kinetic model, was inversely proportional to agonist concentration. The ratio of the rate constants of aggregation and deaggregation was receptor occupancy-dependent and directly proportional to aggregation. Consequently, platelet aggregation was proportional, and deaggregation inversely proportional, to ADP receptor occupancy. We propose that the response of PRP to ADP and to 2-MeS-ADP (2-methylthioadenosine-diphosphate), in vitro, consists of at least two active, concurrent processes, aggregation and deaggregation. Incremental occupancy of the P2T ADP receptor subtype attenuates deaggregation and governs the balance between these two processes.
Maayani
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Department of Anesthesiology, The Mount Sinai School of Medicine, New York, NY 10029-6574, USA. saul.maayani@mssm.edu
Tagliente
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TM
Schwarz
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Martinelli
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Martinez
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Shore-Lesserson
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D000244:Adenosine Diphosphate / Q000031:analogs & derivatives* / Q000494:pharmacology*; D000328:Adult; D020013:Calcium Signaling / Q000187:drug effects; D002451:Cell Compartmentation; D005260:Female; D005343:Fibrinolytic Agents / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D008297:Male; D008565:Membrane Proteins; D008875:Middle Aged; D008954:Models, Biological; D010974:Platelet Aggregation / Q000187:drug effects / Q000502:physiology*; D058910:Purinergic P2 Receptor Agonists; D018048:Receptors, Purinergic P2 / Q000145:classification / Q000502:physiology*; D058925:Receptors, Purinergic P2Y12; D013873:Thionucleotides / Q000494:pharmacology
D005343:Fibrinolytic Agents; D008565:Membrane Proteins; C548576:P2RY12 protein, human; D058910:Purinergic P2 Receptor Agonists; D018048:Receptors, Purinergic P2; D058925:Receptors, Purinergic P2Y12; D013873:Thionucleotides; C002480:methylthio-ADP; D000244:Adenosine Diphosphate