Heritable variation in metabolic traits is likely to affect fitness. In this study, white-footed mice from wild-derived photoresponsive [R, infertile in short day length (SD)] and non-photoresponsive (NR, fertile in SD) selection lines were maintained under short-day (SD 8Light:16Dark), sub-thermoneutral conditions (22 or 12 °C). Mice had significantly higher levels of food intake and resting metabolic rates (RMR) at low temperature. RMR differed significantly between lines (greater in NR mice). In contrast to previous work under thermoneutral conditions, there was no significant difference in overall activity or average daily metabolic rates (ADMR) of mice from the two lines. Reduced activity may reflect behavioral changes under cooler conditions (e.g., nest building) reducing the overall energetic cost of fertility (for NR mice). There was no significant difference in maximal rate of oxygen consumption ([Formula: see text]) between lines. R mice had significantly greater brown adipose tissue and white abdominal fat mass due to both line and temperature. Reaction norms for intake, resting metabolism (RMR/BMR) and level of activity from current (12 and 22 °C) and previously published data (28 °C) demonstrate independent effects of genetics (line) and environment (temperature) for resting metabolism, but a clear interaction between these for activity. The results suggest that fertility under winter conditions imposes metabolic costs that are related to the level of reproductive development. Under the coldest conditions tested, however, mice that remained fertile in SD reduced activity, ADMR and food requirements, decreasing the differential between selection lines. Heritable variation in reaction norms suggests a genetic by environment effect that could be subject to selection.