Suckling has been demonstrated to impair the release of pituitary luteinizing hormone (LH) and to prevent the dramatic increase in plasma LH observed following ovariectomy. In the present study, the effect of suckling (10 pups/animal) for either 1 or 3 weeks on the relative amount of luteinizing hormone releasing hormone (LHRH) present in the hypothalamus and preoptic area of ovariectomized and intact rats was examined using immunocytochemical methodology. Controls consisted of nonlactating animals which were either intact (diestrous) or ovariectomized for 1 or 3 weeks. Brains were removed following transcardial perfusion of phosphate-buffered formalin and Bouin's fixative. After dehydration, clearing and paraffin embedding, the brains were sectioned and LHRH localized by an indirect immunoperoxidase technique. A positive reaction denoting the presence of immunoreactive LHRH was observed over axons and termini throughout the rostral to caudal extent of the median eminence (ME) and surrounding the organum vasculosum of the lamina terminalis (OVLT) in the preoptic area. Ovariectomy resulted in a progressive decline in the concentration of LHRH within the ME as evidenced by a reduction in the intensity of the staining reaction and in the number of axons over which the reaction was observed. In contrast, brains from ovariectomized rats which had been suckled appeared to have concentrations of LHRH in the ME equal to or greater than that of the diestrous controls. Similarly, the concentrations of LHRH In the ME of intact, suckled rats did not differ significantly from that of the diestrous controls. Neither ovariectomy nor suckling produced any observable change in the relative concentration of LHRH located near the OVLT. These data demonstrate that suckling prevents the depletion of LHRH from the ME following ovariectomy and provide evidence for mechanism by which the suckling stimulus may suppress plasma LH.