Heterogeneity of human platelet density subpopulations in aggregation, secretion of ATP, and cytosolic-free calcium concentration
Abstract
AIM: To investigate thrombin (500 U.L-1)-, ADP (0.1-30 mumol.L-1)-, and 5-hydroxytryptamine (5-HT, 3 mumol.L-1)-induced aggregation, secretion of ATP and cytosolic-free calcium mobilization in density subpopulations of human washed platelets.
METHODS: Using Percoll discontinuous gradient.
RESULTS: The human platelets were separated into high density (HD), intermediate density (ID), and low density (LD) subpopulations, and their sizes were diminished with decreasing density (r = 0.978, P < 0.01). The magnitude of aggregations by thrombin, ADP, and 5-HT was more significant in HD platelets than that in LD platelets (P < 0.01). The amount of secretion of ATP induced by thrombin and ADP in HD platelets was also much higher than that in LD platelets (P < 0.01), except for 5-HT which did not cause the ensuring release reaction in any subpopulation of human platelets. Thrombin (1500 U.L-1)-, ADP (mumol.L-1)-, and 5-HT (3 mumol.L-1)-induced cytosolic-free calcium mobilization was evaluated as well. Results showed that the resting level of cytosolic-free calcium concentration ([Ca2+]i) was the same in all subpopulations, about 80-90 nmol.L-1. However, the level of [Ca2+]i mobilization was entirely different, heightened with increasing density.
CONCLUSION: The function of HD platelets was much stronger and more active than that of LD platelets in human.
Keywords:
METHODS: Using Percoll discontinuous gradient.
RESULTS: The human platelets were separated into high density (HD), intermediate density (ID), and low density (LD) subpopulations, and their sizes were diminished with decreasing density (r = 0.978, P < 0.01). The magnitude of aggregations by thrombin, ADP, and 5-HT was more significant in HD platelets than that in LD platelets (P < 0.01). The amount of secretion of ATP induced by thrombin and ADP in HD platelets was also much higher than that in LD platelets (P < 0.01), except for 5-HT which did not cause the ensuring release reaction in any subpopulation of human platelets. Thrombin (1500 U.L-1)-, ADP (mumol.L-1)-, and 5-HT (3 mumol.L-1)-induced cytosolic-free calcium mobilization was evaluated as well. Results showed that the resting level of cytosolic-free calcium concentration ([Ca2+]i) was the same in all subpopulations, about 80-90 nmol.L-1. However, the level of [Ca2+]i mobilization was entirely different, heightened with increasing density.
CONCLUSION: The function of HD platelets was much stronger and more active than that of LD platelets in human.