The current paper presents our initial efforts to establish an in vitro spinal preparation for investigating locomotor pattern generation in mice. We have characterized the step cycle timing from EMG activity in the gastrocnemius (G) and tibialis anterior (TA) muscles of freely moving intact adult as well as neonatal mice and then compared those data with rhythmic EMG activity in an isolated spinal cord-hindlimb preparation. The motor output during the first four days of life was evaluated in an effort to identify the optimal post-partum period for in vitro locomotor studies. The in vitro pattern generating capabilities of the lumbosacral region were tested in both nonhemisected and hemisected preparations. Spontaneous as well as NMDA evoked in vitro activity in the antagonist set of hindlimb muscles included sequences of: 1) synchronous bursting; 2) mixed synchrony and alternation; and/or 3) irregular alternations. The alternating bursting observed in vitro was more often an alternation of sequences rather than a cycle-to-cycle phasing between G and TA muscles. In summary, while there was evidence of reciprocal inhibition in neonates, the circuitry for cycle-to-cycle alternation between antagonists was found to be labile.
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