The effect of stimulating the dorsal raphe nucleus (DRN) on the activity of single relay neurons in the dorsal lateral geniculate nucleus (LGNd) was studied in rats anesthetized with urethane. The position of stimulating electrodes was confirmed on histological sections processed with NADPH-diaphorase histochemistry which could delineate the DRN clearly. During repetitive stimulation of the DRN at 200 Hz for several to 10 seconds no consistent change in firing was observed, but between several and several tens of seconds after the cessation of stimulation spontaneous firing of LGNd neurons was suppressed. In many cases the suppression proceeded concomitantly with augmentation of slow waves in the cortical EEG. The suppression was mimicked by ionophoresis of serotonin, and antagonized by a serotonergic antagonist, methysergide. In addition, in animals in which DRN stimulation became ineffective after it was evoked many times, the suppression could be restored by intraperitoneal administration of a serotonin precursor, 5-hydroxytryptophan. Compilation of peristimulus time histograms revealed that a brief DRN stimulation (5 shocks at 1000 Hz) could also elicit the suppression lasting from 60 to 100 ms or longer after the shocks. These results suggest that serotonin released from terminals of DRN neurons exerts long-latency and long-lasting inhibition of LGNd relay neurons. Along with brainstem noradrenergic and cholinergic systems, the serotonergic projection from the DRN acts to control excitation levels of the forebrain.