The gut-brain axis is of great importance in the control of energy homeostasis. The identification of uroguanylin (UGN), an intestinal released peptide regulated by nutritional status and with anorectic actions, as the endogenous ligand for guanylyl cyclase 2C (GUCY2C) receptor, has revealed a new system in the regulation of energy balance. We show that chronic central infusion of uroguanylin (UGN) reduces weight gain and adiposity in diet-induced obese mice. These effects were independent of food intake and involved specific efferent autonomic pathways. On one hand, brain UGN induces brown adipose tissue thermogenesis as well as browning and lipid mobilization in white adipose tissue through stimulation of the sympathetic nervous system. On the other hand, brain UGN augments fecal output through the vagus nerve. These findings are of relevance as they suggest that the beneficial metabolic actions of UGN through the sympathetic nervous system do not involve non-desirable gastrointestinal adverse effects, such as diarrhea. The present work provides mechanistic insights on how UGN influences energy homeostasis and suggests that UGN action in the brain represents a feasible pharmacological target in the treatment of obesity.