The post-copulatory sexually refractory stage in the male cricket Gryllus bimaculatus consists of the two substages: the first refractory stage (RS1, time-variable) between copulation and spermatophore preparation, and the second refractory stage (RS2, time-constant) between spermatophore preparation and the recommencement of courtship. To understand the mechanism of the timer for RS2, subcuticular or intraganglionic injection of biogenic amines (10(-2) mol l(-1)) was performed immediately after spermatophore preparation. RS2 was shortened by octopamine, 5-HT, 5-HTP and NA-5-HT. Among these, 5-HTP was most potent. It shortened RS2 to maximally about 38% of the control. The shortening effect continued for 4.5 h after subcuticular injection even when the hemolymph was washed out with saline at 1 hour after injection. Simultaneous injection of 5-HTP with the inhibitor (NSD-1015) of 5-HT synthesis enzyme nullified the effect of 5-HTP, indicating that the shortening effect was caused by 5-HT synthesized from extrinsic 5-HTP. Injection of the inhibitor (CHX) of protein synthesis had no effect of on RS2. These results suggest that the reproductive timer in the TAG may be controlled by 5-HT or a second messenger mediated by 5-HT.