It is well established that hypothalamic galanin- and beta-endorphin-containing circuits play important roles in the neuroendocrine regulation of pituitary hormone secretion and sexual behaviors, as well as in feeding. Recent experimental evidence suggests that an opiate-galanin interaction may be involved in these neuroendocrine responses. In particular, galanin and beta-endorphin have been shown to stimulate prolactin release from the pituitary, and concurrently, evoke feeding in the rat. The present study was designed to elucidate the morphological component underlying these responses in the hypothalamus. Sections of the mediobasal hypothalamus of colchicine-pretreated female rats were double immunostained for galanin and beta-endorphin. A dark blue nickel ammonium sulfate-intensified diaminobenzidine reaction was used to visualize galanin profiles, while beta-endorphin neurons were labeled with a light brown diaminobenzidine reaction. Light microscopy revealed putative connections between galanin boutons and beta-endorphin cells. Electron microscopic examination showed that galanin boutons form axo-somatic and axo-dendritic synaptic connections with beta-endorphin neurons. The vast majority (89.6%) of the beta-endorphin-immunoreactive neurons were found to be contacted by galanin-immunopositive fibers in the hypothalamus. To determine the origin of the galanin fibers innervating this region, the arcuate nuclei of additional rats were isolated unilaterally using a Halász-knife. After a ten day survival period, immunostaining was carried out for galanin. The relative surface occupied by galanin immunoreactive profiles on the ipsi- and contralateral sides were compared using an image analyzer. This analysis revealed that deafferentation of the arcuate nucleus did not decrease the density of galanin immunoreactive profiles on the isolated side of the arcuate nucleus compared to the control side, thus, indicating that the galanin boutons contacting beta-endorphin cells are most probably of local origin. These studies support the proposal that galanin-evoked prolactin secretion and feeding behavior may, in part, be mediated by enhanced beta-endorphin release and raises the possibility that a hypothalamic galanin-beta-endorphin axis may operate in the control of other pituitary hormones.