The effects of discrete lesions of the suprachiasmatic and medial preoptic nucleus on luteinizing hormone-releasing hormone (LHRH) neurons of the female rat were examined. The lesions disrupted the estrous cycle and prevented the preovulatory surge of luteinizing hormone and prolactin. Two to three months following the lesions, control and lesioned animals were perfused, the brains were sectioned, and the tissue processed for LHRH immunocytochemistry using the peroxidase antiperoxidase method and a high-titer, conformational antiserum to LHRH. Faintly stained LHRH cells were observed in the preoptic area and the basal hypothalamus at all stages of the estrous cycle. The number of immunoreactive cell bodies varied from a high of 583 cells on proestrus, to a low of 35 cells on estrus (mean ± SEM = 323 ± 59; n = 11). In contrast, the constant estrous animals with lesions showed increased intensity and number of LHRH neurons rostral, lateral and caudal to the lesion. The total number of cells ranged from 625 to 954 cells per animal (mean ± SEM = 784 ± 44; n = 8; p< 0.001 vs. controls). Moreover, all lesioned animals exhibited intense fiber stain in the median eminence region. In conclusion, these data support the hypothesis that persistent estrus is caused by destruction of neurons which directly or indirectly control LHRH neur