The key role of the suprachiasmatic nucleus in the diurnal regulation of anterior pituitary hormone secretions, including PRL, is well established. However, the pathway via suprachiasmatic signals reach the pituitary is ill defined. To determine whether suprachiasmatic efferents innervate neuroendocrine cells, the anterograde tracer, Phaseolus vulgaris leukoagglutinin, was injected iontophoretically into the suprachiasmatic nucleus in parallel with ip administration of fluorogold (20 mg/BW in saline). After visualization of anterogradely labeled processes with a dark blue chromogen. Vibratome sections were immunostained for fluorogold. As fluorogold labeling resulted in dense immunopositive granules without diffuse cytoplasmic labeling, selected sections were further immunostained for cytoplasmic tyrosine hydroxylase (dopamine). Anterogradely labeled suprachiasmatic efferents were observed in the medial preoptic area, periventricular regions, and the lateral aspects of the arcuate and ventromedial nuclei of the hypothalamus, whereas the median eminence and organum vasculosum laminae terminalis lacked labeled suprachiasmatic projections. All of the aforementioned regions contained a high number of cells immunoreactive for fluorogold. However, immunolabeling for fluorogold revealed no retrogradely labeled (ergo neuroendocrine) cells in the suprachiasmatic nucleus. Retrogradely labeled cells in all of these hypothalamic sites, with the exception of the median eminence and organum vasculosum laminae terminalis, were targets of suprachiasmatic nucleus axon terminals. In the preoptic area, anterior hypothalamus, periventricular area, and arcuate nucleus, subpopulations of dopamine cells were retrogradely labeled. In all of these areas, both retrogradely labeled and nonlabeled dopamine cells were frequently found to be in contact with dark blue, anterogradely labeled suprachiasmatic efferents. Electron microscopic examination confirmed the putative connections to be synaptic. This experiment provided evidence that the circadian pacemaker suprachiasmatic nucleus sends efferents onto neuroendocrine cells, but has no contacts with fenestrated capillaries. It was found that a population of median eminence-projective cells targeted by suprachiasmatic axons in the hypothalamus contains dopamine. These observations indicate no direct effect of the circadian pacemaker on the anterior hypophysis, but offer an indirect pathway via circadian signals, mediated by hypothalamic neural systems, that may regulate pituitary hormone secretion, in particular PRL.