Sphingolipid metabolism during human platelet activation

Thrombin activation of human platelets causes release of sphingosine-1-phosphate from platelets and an increase in sphingosine levels. Sphingosine-1-phosphate is also known to potentiate platelet aggregation. Thus, these sphingolipids may serve as second messengers during platelet activation making it possible that another sphingolipid, ceramide, might play a role in platelet function. Platelets are known to contain sphingomyelinase activity and hydrolysis of sphingomyelin by this enzyme yields phosphocholine and ceramide. Since ceramide is thought to exert its effects through regulation of protein kinases and phosphatases, both of which are involved in platelet function, it is possible that ceramide produced during platelet activation could be involved in regulating signal transduction events. To investigate this possibility, potential changes in levels of ceramide and sphingomyelin in resting and thrombin-activated platelets have been evaluated. Thin-layer chromatographic analysis of the total mass or of radiolabeled (14C-palmitate or 14C-serine) pools of ceramide and sphingomyelin did not reveal any significant changes in the concentrations of either of these molecules during platelet activation. In addition, activation of platelets labeled with [14C-choline]-sphingomyelin did not lead to production of 14C-phosphocholine, suggesting that platelet activation did not cause sphingomyelinase to hydrolyze the [14C-choline]-sphingomyelin. Taken together, our results suggest that ceramide does not serve as a second messenger during platelet aggregation.