Characterization of 5-hydroxytryptamine binding sites in the plasma membrane of pig blood platelets.

Plasma membranes were isolated from pig platelets after glycerol facilitated lysis by sucrose density gradient centrifugation. The purity of the membrane fraction was followed by electron microscopy, gel electrophoresis and analysis of acid phosphatase (EC 3.1.3.2) and phosphodiesterases (EC 3.1.4.1). (3H)5-Hydroxytryptamine ([3H]5-HT) was bound to two saturable binding sites of the membranes. The KD value for the high affinity sites was 0.85 nM and for the low affinity sites 0.48 microM. With the exception of tryptamines little or no (3H)5-HT was displaced by serotonin antagonists and uptake inhibitors suggesting another type of binding than that of 5-HT1. Apparently, enhancement of binding in the presence of Na+ was due to stimulation of an uptake process. Binding of (3H)ketanserin and (3H)LSD to pig platelet membranes showed the characteristics of 5-HT2 binding sites previously identified in rat brain. Since ketanserin inhibited 5-HT induced aggregation of pig platelets (IC50 = 14.2 nM), the ketanserin binding sites can be classified as 5-HT2 receptors. The functional properties of these binding sites and their density in pig platelets as compared with brain membranes may motivate studies on 5-HT2 receptors in pig platelets as models for those in nerve endings.