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 Blood, 1 April 2005, Vol. 105, No. 7, pp. 2749-2756.
Prepublished online as a Blood First Edition Paper on November 16, 2004; DOI 10.1182/blood-2004-07-2821.

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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY

Differential requirements for calcium and Src family kinases in platelet GPIIb/IIIa activation and thromboxane generation downstream of different G-protein pathways

Robert T. Dorsam, Soochong Kim, Swaminathan Murugappan, Swathi Rachoor, Haripriya Shankar, Jianguo Jin, and Satya P. Kunapuli

From the Department of Pharmacology, the Department of Physiology, and The Sol Sherry Thrombosis Research Center, Temple University School of Medicine, Philadelphia, PA.

G12/13 or Gq signaling pathways activate platelet GPIIb/IIIa when combined with Gi signaling. We tested whether combined Gi and Gz pathways also cause GPIIb/IIIa activation and compared the signaling requirements of these events. Platelet aggregation occurred by combined stimulation of Gi and Gz pathways in human platelets and in P2Y1-deficient and G{alpha}q-deficient mouse platelets, confirming that the combination of Gi and Gz signaling causes platelet aggregation. When Gi stimulation was combined with Gz stimulation, there was a small mobilization of intracellular calcium. Chelation of intracellular calcium decreased the extent of this platelet aggregation, whereas it abolished the Gq plus Gi-mediated platelet aggregation. Costimulation of Gi plus Gz pathways also caused thromboxane generation that was dependent on outside-in signaling and was inhibited by PP2, a Src family tyrosine kinase inhibitor. Src family tyrosine kinase inhibitors also inhibited platelet aggregation and decreased the PAC-1 binding caused by costimulation of Gi and Gz signaling pathways in aspirin-treated platelets. However, Src family kinase inhibitors did not affect Gq plus Gi-mediated platelet aggregation. We conclude that the combination of Gi plus Gz pathways have different requirements than Gq plus Gi pathways for calcium and Src family kinases in GPIIb/IIIa activation and thromboxane production.


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