Stimulation of the brainstem reticular formation evokes locomotor activity in embryonic chicken (in ovo)

This study was designed to examine the period of embryonic chick development during which descending brainstem-spinal projections, originating from defined avian brainstem locomotor regions, become functionally active. Locomotor activity was examined using a new in ovo preparation for the focal electrical stimulation of embryonic brainstem locomotor regions. Embryos or hatchlings were anesthetized and mounted in a stereotaxic apparatus. Leg and wing muscle electromyographic (EMG) recordings were used to monitor any brainstem-stimulated motor activity. At present, we have been successful in demonstrating coordinated brainstem-evoked locomotion in embryos as early as embryonic day 15. The patterns of evoked locomotor activity were similar to locomotion evoked in hatchling chicks and were of 4 types: (1) alternating hindlimb movements ('stepping'), (2) synchronous (in-phase) hindlimb movements ('hatching'), (3) synchronous wing movements ('flapping'), and (4) simultaneous 'stepping' and 'flapping'. The cycle durations of evoked embryonic hindlimb movements are shorter than those observed for hatchling chicks. The present results are the first direct demonstration of functional connections between descending supraspinal neurons and spinal locomotor circuits at such an early stage of embryonic development. With modifications in technique, it may be possible to demonstrate functional connections at even earlier stages of embryonic development.