Injection of AMP

Mice were fasted overnight and an intraperitoneal injection with the desired dose of 5’-AMP in sterile PBS was given early in the morning to induce the 5’-AMP Induced Deep Hypometabolic (AIHM) state. The metabolic chamber was maintained at ~15°C during these experiments in order to facilitate the induction and maintenance of the AIHM state. The animals were monitored via the CLAMS computer readout and were removed from the metabolic chamber and placed in their room temperature cage upon arousal from the AIHM state. We defined the AIHM state as occurring when an animal’s VO₂ falls below 1200 mL/kg/h, since the mice tend to enter the arousal state when their VO2 rise above 1200 mL/kg/h. We have observed that most mice begin arousal and rapid re-warming not long after reaching a VO₂ of 1500 mL/kg/h.