Neuropeptide FF (FMRFamide-like peptide, morphine-modulating peptide) is an octapeptide isolated from the bovine brain. There is evidence that neuropeptide FF participates in the modulation of nociceptive mechanisms. Neuropeptide FF acts through its own receptors which are distinct from the opiate receptors. In the rat brain neuropeptide FF is found in two major cell populations. We have studied the efferent connections of the hypothalamic neuropeptide FF-containing cell group, which is located in the medial hypothalamus between the dorsomedial, ventromedial and periventricular hypothalamic nuclei. By using an anterograde tracing method (Phaseolus vulgaris leucoagglutinin) combined with double-staining immunohistochemistry we characterized the connections of this cell group with the limbic system, certain hypothalamic nuclei, periaqueductal gray and with the solitary tract nucleus. In the limbic system, the major targets were the lateral septal nucleus, bed nucleus of stria terminalis and certain subnuclei in the amygdala. These connections suggest that neuropeptide FF may act, in addition to its well-characterized action in the sensory system, in limbic functions. Efferent connections to the periaqueductal gray suggest that neuropeptide FF may modulate the opiate mediated analgesia at this site. Good correlation between our results and receptor autoradiography support the idea that the terminal areas which our results show are target areas of the neuropeptide FF-containing system.