Osmotic minipumps (0.25μL/h, Alzet) were aseptically filled with 0.45 μm-filtered sterile phosphate-buffered saline (PBS) or leptin (1.2mg/mL). Minipumps were primed by incubation at 37°C in sterile 0.9% saline for 8 to 16h. Male 7–8 week old TORC1 −/− mice and wild-type littermates were anesthetized under isoflurane and the minipump was implanted under the dorsal skin between the scapulae. Body weight and food intake were measured daily for 10 days, starting after the implantation of the minipumps. After 10 days of infusion, mice were anesthetized and transcardially perfused for brain immunohistochemistry, as described below.
