Directed functional connectivity matures with motor learning in a cortical pattern generator

Sequential motor skills may be encoded by feedforward networks that consist of groups of neurons that fire in sequence (Abeles 1991; Long et al. 2010). However, there has been no evidence of an anatomic map of activation sequence in motor control circuits, which would be potentially detectable as directed functional connectivity of coactive neuron groups. The proposed pattern generator for birdsong, the HVC (Long and Fee 2008; Vu et al. 1994), contains axons that are preferentially oriented in the rostrocaudal axis (Nottebohm et al. 1982; Stauffer et al. 2012). We used four-tetrode recordings to assess the activity of ensembles of single neurons along the rostrocaudal HVC axis in anesthetized zebra finches. We found an axial, polarized neural network in which sequential activity is directionally organized along the rostrocaudal axis in adult males, who produce a stereotyped song. Principal neurons fired in rostrocaudal order and with interneurons that were rostral to them, suggesting that groups of excitatory neurons fire at the leading edge of travelling waves of inhibition. Consistent with the synchronization of neurons by caudally travelling waves of inhibition, the activity of interneurons was more coherent in the orthogonal mediolateral axis than in the rostrocaudal axis. If directed functional connectivity within the HVC is important for stereotyped, learned song, then it may be lacking in juveniles, which sing a highly variable song. Indeed, we found little evidence for network directionality in juveniles. These data indicate that a functionally directed network within the HVC matures during sensorimotor learning and may underlie vocal patterning.