Thalamus drives vocal onsets in the zebra finch courtship song.

While motor cortical circuits contain information related to specific movement parameters1, long-range inputs also have a critical role in action execution2,3. Thalamic projections can shape premotor activity2-6 and have been suggested7 to mediate the selection of short, stereotyped actions comprising more complex behaviours8. However, the mechanisms by which thalamus interacts with motor cortical circuits to execute such movement sequences remain unknown. Here we find that thalamic drive engages a specific subpopulation of premotor neurons within the zebra finch song nucleus HVC (proper name) and that these inputs are critical for the progression between vocal motor elements (that is, 'syllables'). In vivo two-photon imaging of thalamic axons in HVC showed robust song-related activity, and online perturbations of thalamic function caused song to be truncated at syllable boundaries. We used thalamic stimulation to identify a sparse set of thalamically driven neurons within HVC, representing ~15% of the premotor neurons within that network. Unexpectedly, this population of putative thalamorecipient neurons is robustly active immediately preceding syllable onset, leading to the possibility that thalamic input can initiate individual song components through selectively targeting these 'starter cells'. Our findings highlight the motor thalamus as a director of cortical dynamics in the context of an ethologically relevant behavioural sequence.

Sleep replay reveals premotor circuit structure for a skilled behavior.

Neural circuits often exhibit sequences of activity, but the contribution of local networks to their generation remains unclear. In the zebra finch, song-related premotor sequences within HVC may result from some combination of local connectivity and long-range thalamic inputs from nucleus uvaeformis (Uva). Because lesions to either structure abolish song, we examine "sleep replay" using high-density recording methods to reconstruct precise song-related events. Replay activity persists after the upstream nucleus interfacialis of the nidopallium is lesioned and slows when HVC is cooled, demonstrating that HVC provides temporal structure for these events. To further gauge the importance of intra-HVC connectivity for shaping network dynamics, we lesion Uva during sleep and find that residual replay sequences could span syllable boundaries, supporting a model in which HVC can propagate sequences throughout the duration of the song. Our results highlight the power of studying offline activity to investigate behaviorally relevant circuit organization.