Acute stress affects gut functions through the activation of corticotropin-releasing factor (CRF) receptors. The impact of acute stress on pelvic viscera in the context of chronic stress is less characterized. We investigated the colonic, urinary and locomotor responses monitored as fecal pellet output (FPO), urine voiding and ambulatory activity in female and male CRF-overexpressing (CRF-OE) mice, a chronic stress model, and their wild type littermates (WTL). Female CRF-OE mice, compared to WTL had enhanced FPO to 2-min handling (150%) and 60-min novel environment (155%), while displaying a similar response to a 60-min partial restraint stress. Female CRF-OE mice, compared to WTL, also had a significantly increased number of urine spots (7.3 ± 1.4 vs. 1.3 ± 0.8 spots/h) and lower locomotor activity (246.8 ± 47.8 vs. 388.2 ± 31.9 entries/h) to a novel environment. Male CRF-OE mice and WTL both responded to novelty, but failed to show differences in colonic and locomotor responses. Male WTL, compared to female WTL, had higher FPO (113%). In female CRF-OE mice, the CRF₁/CRF₂ receptor antagonist, astressin B, and the selective CRF₂ receptor agonist, mUcn 2 injected peripherally, prevented the enhanced defecation without affecting urine or locomotor responses to novelty. RT-PCR showed that CRF₁ and CRF₂ receptors are expressed in the mouse colonic tissues. The data show that chronic stress, due to continuous central CRF-overdrive, renders female CRF-OE mice to have enhanced pelvic and behavioral responses to superimposed mild stressors, and that CRF₁-initiated colonic response can be counteracted by selective activation of peripheral CRF₂ receptor.