The hypocretins (Hcrt; also known as orexins) and melanin-concentrating hormone comprise distinct families of neuropeptides synthesized in cells located in the lateral hypothalamus and adjacent areas. The Hcrts are thought to modulate food intake and sleep/wake patterns in mammals. Melanin-concentrating hormone has a well-documented role in energy metabolism. A moderate plexus of Hcrt immunoreactive terminals has been described in the hamster intergeniculate leaflet, part of the circadian rhythm system. This study investigated the origin of Hcrt-immunoreactive terminals in the intergeniculate leaflet and determined whether melanin-concentrating hormone neurons also project to the intergeniculate leaflet. The tracer, cholera toxin beta-subunit, was injected into the intergeniculate leaflet of the golden hamster. Double-label fluorescent immunohistochemistry for cholera toxin beta-subunit and Hcrt or melanin-concentrating hormone was then performed to identify retrogradely labeled cells also containing immunoreactive peptide. Most cholera toxin beta-subunit-labeled cells were detected in the medial zona incerta and sub-incertal zone, with few observed in the lateral hypothalamus. Hcrt-immunoreactive cells were abundant and found largely in the lateral hypothalamus and adjacent nuclei. Melanin-concentrating hormone cells were also abundant in the medial zona incerta, in close proximity to cholera toxin beta-subunit-labeled cells, but ventral to them. Cells containing both cholera toxin beta-subunit and Hcrt-immunoreactive, were present in the dorsal aspect of the lateral hypothalamus. The number observed was small, < or = 1% of the total number of Hcrt cells counted in the hamster. No cholera toxin beta-subunit-immunoreactive cells also contained melanin-concentrating hormone and no melanin-concentrating hormone-immunoreactive processes were evident in the intergeniculate leaflet. The results show that a small number of lateral hypothalamus cells containing Hcrt-immunoreactivity project to the intergeniculate leaflet, but they are scattered rather than collected into a discrete group. At the present time there is no information regarding the function of these cells, although they may contribute to the regulation of sleep/arousal, circadian rhythmicity, or vestibulo-oculomotor function.