Brown adipose tissue thermogenesis is an important component of energy expenditure as exemplified in rodents. Other tissues such as white adipose tissue and muscle are also capable of thermogenesis, but regulation of heat production in these tissues is poorly understood. We employed a relatively large animal model, the ovariectomized sheep, in which site-specific temperature measurements were made as an index of thermogenic output. Dataloggers were implanted into the retroperitoneal (visceral) fat, the gluteal (subcutaneous) fat and the skeletal muscle of the hind limb, and were programmed to record temperature every 15 min. Animals (n = 4) were then placed on a feeding schedule (fed between 1100 and 1600 h) to entrain a post-prandial response in thermogenesis. Baseline thermogenesis (0800–1100 h) was higher (P < 0.05) in visceral fat and muscle than in gluteal fat, whereas the amplitude of the postprandial rise was similar at all three sites. Intracerebroventricular infusion into the lateral ventricle of either vehicle (artificial cerebrospinal fluid) or leptin (10 µg/h at 100 µl/h) for 24 h (0900–0900 h) was performed in a cross-over design with a 1-wk recovery period between treatments. Central leptin infusion did not alter the basal thermogenic rate but markedly enhanced the postprandial response in both fat and muscle tissues. This was manifest by increased (P < 0.05) amplitude and duration of the postprandial thermogenic response and the effect was greater in muscle and visceral fat than in gluteal fat. These data demonstrate that leptin is able to regulate thermogenesis in muscle, providing a novel target for the manipulation of energy balance.