Modulation and pre-amplification of PAR1 signalling by ADP acting via the P2Y12 receptor during platelet subpopulation formation

Shakhidzhanov, S.S.; Shaturny, V.I.; Panteleev, M.A.; Sveshnikova, A.N.

Авторы: Shakhidzhanov S.S., Shaturny V.I., Panteleev M.A., Sveshnikova A.N.
Журнал: Biochimica et Biophysica Acta - General Subjects
Том: 1850
Номер: 12
Год издания: 2015
Издательство: Elsevier BV
Местоположение издательства: Netherlands
Первая страница: 2518
Последняя страница: 2529
DOI: 10.1016/j.bbagen.2015.09.013
Аннотация: Background Two major soluble blood platelet activators are thrombin and ADP. Of these two, only thrombin can induce mitochondrial collapse and programmed cell death leading to phosphatidylserine (PS) exposure required for blood clotting reactions acceleration. Thrombin can also greatly potentiate collagen-induced PS exposure. However, ADP acting through the P2Y12 receptor was shown to increase the PS-exposing (PS+) platelets fraction produced by thrombin or thrombin-plus-collagen via an unknown mechanism. Methods We developed a comprehensive multicompartmental computational model of platelet PAR1-and-P2Y12 calcium signal transduction that included cytoplasmic signalling, dense tubular system and mitochondria. To test model predictions, flow cytometry experiments with washed, annexin V-labeled platelets were performed. Results Stimulation of thrombin receptor PAR1 in the model induced cytoplasmic calcium oscillations, calcium uptake by mitochondria, opening of the permeability transition pore and collapse of the mitochondrial membrane potential. ADP stimulation of P2Y12 led to cAMP decrease that, in turn, caused changes in phospholipase C phosphorylation by protein kinase A, increase in cytoplasmic calcium level and, consequently, PS+ platelet formation. ADP addition before stimulation of PAR1 produced much greater increase of the PS+ fraction because cAMP concentration had time to go down prior to calcium oscillations; this prediction was also tested and confirmed experimentally. Conclusion These results suggest a mechanism of ADP-dependent PS exposure regulation and show a likely mode of action that could be important for the PS exposure regulation in thrombi, where ADP is released before thrombin formation.
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Modulation and pre-amplification of PAR1 signalling by ADP acting via the P2Y12 receptor during platelet subpopulation formationстатья