We report studies of neurotransmitter and receptor species mediating the inhibition of LH release in ovariectomized (OVX) rats and stimulation in OVX estrogen-primed rats induced by electrical stimulation of the dorsal raphe nucleus (DRN). Attempts were made to block effects of stimulation of the DRN in groups of four to nine ovariectomized rats with or without priming with estradiol benzoate (EB) by pretreatment with the 5HT receptor antagonists ketanserin, methysergide, or metergoline; the a-adrenergic antagonist phenoxybenzamine; or the opiate antagonist naltrexone. Blood samples were collected via jugular cannulae from the unanesthetized rats at 10-min intervals before and during electrical stimulation of the DRN, and assayed by double antibody radioimmunoassay for LH. To test the effect of stimulation, data from animals in each treatment group were subjected to analysis of variance to obtain the contrast matrix including prestimulation and stimulation time points of interest. The contrast matrix was then used to construct an F ratio and thereby to evaluate the level of significance. In unprimed OVX rats the sustained decrease in plasma LH concentration during stimulation was prevented in rats pretreated with ketanserin or phenoxybenzamine. Methysergide pretreatment delayed the inhibitory effect of DRN electrical stimulation for 30 min, whereas metergoline and naltrexone were ineffective. In EB-primed animals the increase in plasma LH observed during stimulation was prevented in rats pretreated with metergoline, and reversed in those receiving naltrexone. Ketanserin limited the duration of the increase to 10 min, while phenoxybenzamine and methysergide had no significant effect. These results suggest that the inhibitory effect of dorsal raphe stimulation on release of LH in OVX rats may be mediated by SHT, and a-adrenergic systems, whereas the stimulatory effects (OVX + EB) may involve primarily SHT, and opiate receptors.