Thrombin-induced caspases 3 and 9 translocation to the cytoskeleton is independent of changes in cytosolic calcium in human platelets

Apoptosis has been shown to be associated with changes in cytosolic free calcium concentration (Ca(2+)](c)). Here we show that the agonist thrombin induces activation of caspases 9 and 3 and translocation of the caspase active forms and procaspases to the cytoskeleton in human platelets. Dimethyl-BAPTA loading did not affect thrombin-induced caspase 9 and 3 activation or translocation suggesting that these responses are independent of increases in Ca(2+)](c). Treatment with thapsigargin plus ionomycin, to induce extensive Ca(2+) store depletion and subsequent increase in Ca(2+)](c), stimulates caspase activation although it was unable to induce caspase translocation to the cytoskeleton. Similar results were observed in cells loaded with dimethyl-BAPTA, suggesting that activation of caspases 9 and 3 by thapsigargin plus ionomycin does not require rises in Ca(2+)](c). These findings suggest that thrombin-induced caspase 9 and 3 activation and translocation are independent on rises in Ca(2+)](c) but might require store depletion in human platelets.