Outside of the tropics, environmental conditions fluctuate in a generally predictable manner across the year. Many small mammals have evolved mechanisms, such as seasonal breeding and annual adjustments in physiology, morphology, and behavior, that promote winter survival when food is scarce and thermoregulation is challenging. Photoperiod (day length) is a cue used by many seasonal breeders to predict seasonal changes in environmental conditions. One system that is uniquely situated to mediate photoperiod-induced alterations in physiology is the autonomic nervous system (ANS). The 2 branches of the ANS are key regulators of immune responses, thermoregulation, and energy balance, functions that undergo marked shifts in baseline and reactivity following acclimation to short day lengths. Although previous studies have investigated the effects of photoperiod on ANS endpoints, this study examined the direct effects of photoperiod on integrated ANS function. To test the hypothesis that short day lengths increase parasympathetic and sympathetic tones, we maintained adult male Siberian hamsters (Phodopus sungorus) to either long or short photoperiods and then analyzed electrocardiogram recordings. Short day lengths increased both parasympathetic tone, as measured by respiratory sinus arrhythmia, and sympathetic control of the heart, measured with autonomic blockade. Additionally, short day lengths enhanced the withdrawal of parasympathetic control and the increase of sympathetic tone in response to acute restraint stress. Finally, these effects were discovered to be independent of circulating androgens. These data indicate that the ANS of Siberian hamsters undergoes profound changes following prolonged exposure to short winter-like day lengths.