Enhancement of ADP-induced aggregation by 5-HT in rabbit platelets
Abstract
AIM: To study the enhanced effects of 5-hydroxytryptamine (5-HT) on ADP-induced aggregation.
METHODS: Platelet aggregation was quantified by the light transmission, the cytosolic-free calcium ([Ca2+]i) was measured by digital fluorescent microscopy, and inositol 1,4,5-triphosphate (IP3) was determined by receptor binding assay. RESULTS: In rabbit platelet-rich plasma (PRP), 5-HT 0.03-3 mumol.L-1 induced a decrease in light transmission (DLT) in a concentration-dependent manner with centralization of granules, as revealed by electron microscopy. The DLT was accompanied with neither platelet aggregation nor a release reaction. In single washed platelets loaded with Fura-2, 5-HT caused a concentration-dependent elevation of [Ca2+]i, and IP3 level was also transiently increased in washed platelets at 15 s after stimulation by 5-HT. Adenosine diphosphate (ADP) also caused DLT transiently in PRP before its own aggregation without a release reaction. Pretreatment of PRP or washed platelets with 5-HT, the DLT by ADP was reduced concentration-dependently and ADP-induced aggregation and [Ca2+]i mobilization were enhanced.
CONCLUSION: The enhancement of ADP-induced aggregation was attributed to the superimposition of the calcium release from the storage sites and calcium influx induced by ADP over the calcium release from the storage sites by 5-HT.
Keywords:
METHODS: Platelet aggregation was quantified by the light transmission, the cytosolic-free calcium ([Ca2+]i) was measured by digital fluorescent microscopy, and inositol 1,4,5-triphosphate (IP3) was determined by receptor binding assay. RESULTS: In rabbit platelet-rich plasma (PRP), 5-HT 0.03-3 mumol.L-1 induced a decrease in light transmission (DLT) in a concentration-dependent manner with centralization of granules, as revealed by electron microscopy. The DLT was accompanied with neither platelet aggregation nor a release reaction. In single washed platelets loaded with Fura-2, 5-HT caused a concentration-dependent elevation of [Ca2+]i, and IP3 level was also transiently increased in washed platelets at 15 s after stimulation by 5-HT. Adenosine diphosphate (ADP) also caused DLT transiently in PRP before its own aggregation without a release reaction. Pretreatment of PRP or washed platelets with 5-HT, the DLT by ADP was reduced concentration-dependently and ADP-induced aggregation and [Ca2+]i mobilization were enhanced.
CONCLUSION: The enhancement of ADP-induced aggregation was attributed to the superimposition of the calcium release from the storage sites and calcium influx induced by ADP over the calcium release from the storage sites by 5-HT.