Um100a

Under isoflurane anesthesia, the animals were implanted with stainless steel skull screws to serve as recording electrodes. Two Electrodes were used for bipolar recording and were located 3mm anterior to lambda and 4 mm left and right of the medial suture. The third electrode served as reference and was located 1mm anterior to bregma and 1mm to the right of the midline defined by the medial suture. The electrodes were connected to a tri-polar connector (Plastics One, VA) and dental cement was used to cover the electrodes so that only the connector was exposed. Animals were used one to two weeks after electrode implantation. The BioPac Systems MP150 was used to record digital EEG using a BioPac UM100A preamplifier. Sampling rate was 200Hz.

Hindlimb muscle contractile forces were determined as has been previously described (21 (link)). In brief, mice were anesthetized with 40–50mg/kg Pentobarbital and maintained at 37°C using radiant heat. The sciatic nerve was severed to prevent retrograde stimulation. The leg was immobilized with 4-0 silk suture binding the tibia to a stainless steel metal rod mounted on a platform. The third metatarsal was sutured to a TSD 105A force transducer (Biopac Systems) using 4-0 silk. Two Grass E2 platinum subdermal needle electrodes were placed through the Gastrocnemius muscle at a separation distance of 5 mm. Five-volts square waves pulses with duration of 5 ms were delivered at intervals of 500 ms through the electrodes using STM100C (Biopac Systems). The animals were stimulated for 10 min. Tension from the force transducer was recorded with DA100C system and plotted using a UM100A and MP150 system with Acknowledge software (Biopac Systems). Peak force and fatigue force after 10 min of stimulation were measured. Sham animals subjected to anesthesia alone were used for baseline values to which the IR groups were compared.