Food availability affects whether mammals communicate their interest in interacting with opposite-sex conspecifics. This study examined the responses of voles to over-marks, and factors that influence the formation and maintenance of a preference for the top-scent in an over-mark. Specifically, we investigated how food deprivation affected the amount of time male and female voles exposed to an over-mark, later responded to the marks of the top- and bottom-scent donors when subsequently presented with the two scents side by side. Males and females that were not food deprived and males that were food deprived 6 h before exposure to an over-mark later maintained a preference for the donor of the top-scent mark compared with the donor of the bottom-scent mark of the over-mark. Females that were food deprived for 6 h before or after exposure of the over-mark and males food deprived 6 h after the exposure to the over-mark showed no preference for the top-scent mark donor. Re-feeding females that were food deprived for 6 h before exposure to an over-mark was sufficient to restore their preference for the mark of the top-scent male over that of the bottom-scent male. The different responses of food-deprived male and female voles to over-marks of opposite-sex conspecifics may be associated with differences in their tactics for interacting with potential mates and the higher energetic costs of reproduction in female voles than in male voles.