For meadow voles, Microtus pennsylvanicus, seasonal differences exist in self-grooming and in odor preferences for conspecifics, two behaviors which facilitate sexual interactions in this species. Both behaviors are mediated by photoperiodically-induced changes in circulating gonadal steroid hormone titers which, in turn, can be transduced by the duration of the melatonin signal that a seasonally breeding animal receives. The goal of this study was to determine whether exogenous melatonin administration affects circulating gonadal steroid hormone titers in meadow voles, and whether it influences their odor preferences and self-grooming behavior to same- and opposite-sex conspecifics. Long-photoperiod voles that did not receive exogenous melatonin had higher testosterone (males) and estradiol (females) titers than did short-photoperiod voles and long-photoperiod voles treated with melatonin for 12 weeks; the latter had similar estradiol and testosterone titers. Long-photoperiod voles that did not receive melatonin preferred the scent marks of long-photoperiod opposite-sex conspecifics and spent more time self-grooming in response to their odors than those of either long-photoperiod same-sex, short-photoperiod same-sex, or short-photoperiod opposite-sex conspecifics. Long-photoperiod voles that received melatonin, however, no longer preferred the marks of long-photoperiod opposite-sex conspecifics and no longer spent more time self-grooming in response to their odors, not unlike the odor preferences and self-grooming behavior of short-photoperiod voles. As a whole, the data suggest that the duration of the melatonin signal is likely involved in mediating the photoperiodically-induced changes in gonadal steroid hormones that mediate a meadow vole's odor preferences for opposite-sex conspecifics and its self-grooming response to those marks.