Using quantitative autoradiography, 2-I-125-melatonin binding was investigated throughout the Light:dark cycle in the suprachiasmatic nuclei (SCN), paraventricular nuclei (PVT), and pars tuberalis (PT) of adult female Siberian hamsters kept for 10 weeks in either long or short photoperiods (LP or SP, respectively). Plasma melatonin concentrations were measured by radioimmunoassay, and the sexual status of the animals was established by visual inspection of vaginal smears and by weighing uteri after sacrifice. The SCN displayed neither daily nor photoperiod-dependent variations in specific binding. Melatonin receptors in these nuclei would be regulated neither by plasma melatonin nor by the light:dark cycle or sexual steroids. By contrast, melatonin receptor density in the PT displayed both strong daily (maximal values during the first half of the light period and minimal values during the night) and photoperiod-dependent (maximal values in LP) variations. These variations depended on changes in the maximal binding (B-max) without differences in the dissociation constant (K-d). Daily melatonin receptor densities in the PT of LP- and SP-exposed animals might be regulated by the dark:light transition but not by melatonin. Daily profiles of 2-I-125-melatonin-specific binding in the PT were independent of photoperiod. Factors underlying the photoperiod-related variations presently are unknown. Concerning the PVT, weak variations in specific binding were detected in SP only when time points were grouped according to the Light or dark periods. It is not yet possible to conclude whether they have any physiological relevance. These results show clearly that the regulation of melatonin receptors varies among structures (SCN, PVT, and PT) in the Siberian hamster and is also totally different from that found in the rat.