Stimulation of the active transport of serotonin into mouse platelets by the sulfhydryl oxidizing agent diamide

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Abstract
The purpose of this study was to determine the effects of diamide, a reversible sulfhydryl oxidizing agent, on the transport of serotonin (5‐HT) by mouse platelets. Diamide produced a concentration‐dependent (10–200 μM) stimulation of 5‐HT transport that was rapid and sustained over 0–10 minutes of incubation. When platelets were incubated with diamide (10–200 μM) in the presence of glucose, the content of reduced glutathione was significantly decreased only at a final concentration of 200 μM, while washed platelets incubated with diamide (10–200 μM), in the absence of glucose, had a significant concentration‐dependent decrease in their content of reduced glutathione. Fluoxetine, an inhibitor of the platelet 5‐HT transporter, blocked diamide‐induced stimulation of 5‐HT transport. The kinetics of 5‐HT transport showed that diamide caused a marked increase in the maximal rate of transport (Vmax control = 28.4 ± 1.4 vs. Vmax diamide = 60.9 ± 4.1 pM/108 platelets/4 min) but did not significantly alter the Km values. Ouabain, an inhibitor of platelet Na+‐K+ ATPase, blocked the stimulation by diamide in a concentration‐dependent manner. Dithiothreitol, a disulfide reducing agent, was able to partially reverse the stimulation of platelet 5‐HT transport caused by diamide. This study has shown that diamide can stimulate the active transport of 5‐HT by mouse platelets and suggests a possible role for free sulfhydryl groups in the regulation of this process.