Effect of physiologic shear stresses and calcium on agonist-induced platelet aggregation, secretion, and thromboxane...

Platelets in vivo react under low-shear venous-flow as well as high-shear arterial-flow conditions. Because most studies were carried out at low shear stresses, platelet granule secretion at high shear has not been examined thoroughly. We investigated the secretion of all three types of platelet granules and thromboxane A(2) formation at high shear after stimulation with ADP or thrombin.

Washed human platelets were reacted rapidly (<5 s) in a quenched-flow system simulating high-shear arterial-flow conditions (30 dyn/cm(2)). For comparison, we employed a low-shear stirring system (1-5 dyn/cm(2)). Serotonin release and membrane exposure of P-selectin (alpha), CD63 (dense), and CD107a (lysosomes) were used to assess granule secretion. Aggregation was evaluated by resistive-particle counting of the remaining platelet singlets.

ADP and thrombin induced similar strong levels of aggregation ( approximately 70%) at high shear by 5 s. Thrombin also caused release of about 40% of all alpha and dense granules within 5 s. However, by 5 s at high shear, ADP failed to induce significant granule secretion or thromboxane A(2) formation (<5%, p>0.05). By 10 min at low shear, ADP caused secretion and thromboxane A(2) formation only at non-physiological, micromolar extracellular Ca(2+) concentrations. These results emphasize the ability of thrombin to initiate multiple aspects of platelet function within seconds, while ADP was only able to induce rapid aggregation.