Siberian dwarf hamsters form monogamous male-female pair bonds. Disruption of the pair bond results in increases in body mass and behavioral alterations similar to profiles seen in human atypical depression. We examined behavioral and neuroendocrine correlates associated with separation of the male from his mate. Animals were paired (n = 28 pairs) for 3 weeks, then 15 pairs were separated and 13 pairs remained as controls. Behaviors of the males were observed in a novel environment following 3 weeks of pairing and 4 weeks of separation. The 5-min behavioral test monitored exploratory, territorial behavior, and locomotor activity. Separated males showed a significant increase in body mass (p < 0.01). paralleled by an increase in food consumption (p < 0.01). Separated males had decreased seminal vesicle mass (p < 0.05) and testicular mass (p < 0.05). Behavioral analysis revealed that separated males showed no significant differences in grooming, scent-marking, alert on 2 feet, or escape behavior when compared to paired males. Separated males did show significant increases in inactivity (p < 0.05). Plasma cortisol levels were significantly increased in separated animals (p < 0.05), but there were no significant effects on testosterone. Resting levels of plasma norepinephrine and epinephrine were less in separated males, but this was significant only for norepinephrine (p < 0.05). In conclusion, separation stress was accompanied by increased hypothalamic-pituitary-adrenal axis function and decreased peripheral sympathetic nervous system activity and decreased reproductive profiles.