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1. Blood platelets containing different amounts of 5-hydroxytryptamine (5-HT) were produced in vivo by the injection of 5-hydroxytryptamine or of reserpine into normal rabbits and of 5-hydroxytryptamine into reserpinized rabbits. Before and after these injections the aggregation of platelets was measured in vitro.

2. Platelets of untreated rabbits were aggregated by adenosine diphosphate (ADP) and by 5-hydroxytryptamine; (—)-adrenaline alone did not produce aggregation but markedly increased aggregation by 5-hydroxytryptamine.

3. Platelets saturated with 5-hydroxytryptamine in vivo were no longer aggregated in vitro by 5-hydroxytryptamine or by 5-hydroxytryptamine plus adrenaline, but their aggregation by ADP was unchanged.

4. Platelets from reserpinized rabbits lost about 99% of their 5-hydroxytryptamine; the aggregation of these platelets did not differ significantly from that of platelets from control rabbits.

5. Platelets from reserpinized rabbits injected with 5-hydroxytryptamine were aggregated neither by the amine alone nor by 5-hydroxytryptamine plus adrenaline, although these platelets contained much less 5-hydroxytryptamine than saturated platelets and only about one tenth as much as platelets from untreated rabbits.

6. The findings support the hypothesis that the inhibitory effect of 5-hydroxytryptamine administered in vivo on platelet aggregation in vitro is due to the association of the amine with the platelet membrane.