Local Axonal Conduction Shapes the Spatiotemporal Properties of Neural Sequences.

Sequential activation of neurons has been observed during various behavioral and cognitive processes, but the underlying circuit mechanisms remain poorly understood. Here, we investigate premotor sequences in HVC (proper name) of the adult zebra finch forebrain that are central to the performance of the temporally precise courtship song. We use high-density silicon probes to measure song-related population activity, and we compare these observations with predictions from a range of network models. Our results support a circuit architecture in which heterogeneous delays between sequentially active neurons shape the spatiotemporal patterns of HVC premotor neuron activity. We gauge the impact of several delay sources, and we find the primary contributor to be slow conduction through axonal collaterals within HVC, which typically adds between 1 and 7.5 ms for each link within the sequence. Thus, local axonal "delay lines" can play an important role in determining the dynamical repertoire of neural circuits.

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.

Modality-invariant audio-visual association coding in crow endbrain neurons.

Single neuron activity in the corvid nidopallium caudolaterale (NCL), the supposed avian functional analog of the prefrontal cortex, represents associations of auditory with visual stimuli. This is of high adaptive value for songbirds that need to rely on audio-visual associations to communicate, find a mate or escape predators. However, it remains unclear whether NCL neurons can represent cross-modal associations in a modality invariant, abstract fashion. To dissociate between modality-dependent and modality-invariant NCL activity, we trained two crows to match auditory sample cues with visual test stimuli, and vice versa, across a temporal memory delay. During sample presentation, NCL activity selectively encoded associations in a modality invariant fashion. During the delay, we observed subject specific, population-level coding biases in NCL activity. Despite of these biases, task relevant information could be decoded equally well from either subject's neuronal delay activity. Decoding success was facilitated by many mixed selectivity neurons, which mediated high dimensional representations of task variables on the NCL population level. These results parallel findings from the mammalian PFC, suggesting common mechanisms responsible for the adaptability of multimodal association areas across taxa.

Input and Output Connections of the Crow Nidopallium Caudolaterale.

The avian telencephalic structure nidopallium caudolaterale (NCL) functions as an analog to the mammalian prefrontal cortex. In crows, corvid songbirds, it plays a crucial role in higher cognitive and executive functions. These functions rely on the NCL's extensive telencephalic connections. However, systematic investigations into the brain-wide connectivity of the NCL in crows or other songbirds are lacking. Here, we studied its input and output connections by injecting retrograde and anterograde tracers into the carrion crow NCL. Our results, mapped onto a published carrion crow brain atlas, confirm NCL multisensory connections and extend prior pigeon findings by identifying a novel input from the hippocampal formation. Furthermore, we analyze crow NCL efferent projections to the arcopallium and report newly identified arcopallial neurons projecting bilaterally to the NCL. These findings help to clarify the role of the NCL as central executive hub in the corvid songbird brain.

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.

Cross-Modal Associative Mnemonic Signals in Crow Endbrain Neurons.

The ability to associate stimuli across time and sensory modalities endows animals and humans with many of the complex, learned behaviors. For successful performance, associations need to be retrieved from long-term memory and maintained active in working memory. We investigated how this is accomplished in the avian brain. We trained carrion crows (Corvus corone) to perform a bimodal delayed paired associate task in which the crows had to match auditory stimuli to delayed visual items. Single-unit recordings from the association area nidopallium caudolaterale (NCL) revealed sustained memory signals that selectively correlated with the learned audio-visual associations across time and modality, and sustained activity prospectively encoded the crows' choices. NCL neurons carried an internal, stimulus-independent signal that was predictive of error and type of error. These results underscore the role of corvid NCL in synthesizing external multisensory information and internal mnemonic data needed for executive control of behavior.

The long and the short of it: rule-based relative length discrimination in carrion crows, Corvus corone.

Birds and other nonhuman animals can choose the larger of two discrete or continuous quantities. However, whether birds possess the conceptual grasp and cognitive control to flexibly switch between relative more-or-less-than judgments remains elusive. We therefore tested carrion crows in a rule-based line-length discrimination task to flexibly select lines presented on a touchscreen according to their relative length. In the first experiment, the crows needed to discriminate a shorter from a longer line, and vice versa. In the second experiment, the crows were required to choose a medium long line among three lines of different length (intermediate-size task). The crows switched effortlessly between "longer than/shorter than" rules, showing no signs of trial history affecting switching performance. They reliably chose the relatively longer and shorter line length, thus demonstrating a concept of greater than/less than with a continuous magnitude. However, both crows failed to discriminate a line of 'medium' length embedded in longer and shorter lines. These results indicate that relational discrimination exhibits different cognitive demands. While a greater than/less than concept requires only one relational comparison (with the respectively greater or smaller magnitude), the discrimination of a 'medium' magnitude demands to relate two or more comparisons, which might overburden crows and maybe animals in general.