Teresa A. Nick

Modulation of Perineuronal Nets and Parvalbumin with Developmental Song Learning

Neural circuits and behavior are shaped during developmental phases of maximal plasticity known as sensitive or critical periods. Neural correlates of sensory critical periods have been identified, but their roles remain unclear. Factors that define critical periods in sensorimotor circuits and behavior are not known. Birdsong learning in the zebra finch occurs during a sensitive period similar to that for human speech. We now show that perineuronal nets, which correlate with sensory critical periods, surround parvalbumin-positive neurons in brain areas that are dedicated to singing. The percentage of both total and parvalbumin-positive neurons with perineuronal nets increased with development. In HVC (this acronym is the proper name), a song area important for sensorimotor integration, the percentage of parvalbumin neurons with perineuronal nets correlated with song maturity. Shifting the vocal critical period with tutor song deprivation decreased the percentage of neurons that were parvalbumin positive and the relative staining intensity of both parvalbumin and a component of perineuronal nets. Developmental song learning shares key characteristics with sensory critical periods, suggesting shared underlying mechanisms.

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.

Daily and Developmental Modulation of “Premotor” Activity in the Birdsong System

Human speech and birdsong are shaped during a sensorimotor sensitive period in which auditory feedback guides vocal learning. To study brain activity as song learning occurred, we recorded longitudinally from developing zebra finches during the sensorimotor phase. Learned sequences of vocalizations (motifs) were examined along with contemporaneous neural population activity in the song nucleus HVC, which is necessary for the production of learned song (Nottebohm et al. 1976 : J Comp Neurol 165:457–486; Simpson and Vicario 1990 : J Neurosci 10:1541–1556). During singing, HVC activity levels increased as the day progressed and decreased after a night of sleep in juveniles and adults. In contrast, the pattern of HVC activity changed on a daily basis only in juveniles: activity bursts became more pronounced during the day. The HVC of adults was significantly burstier than that of juveniles. HVC bursting was relevant to song behavior because the degree of burstiness inversely correlated with the variance of song features in juveniles. The song of juveniles degrades overnight (Deregnaucourt et al. 2005 : Nature 433:710–716). Consistent with a relationship between HVC activity and song plasticity (Day et al. 2008 : J Neurophys 100:2956–2965), HVC burstiness degraded overnight in young juveniles and the amount of overnight degradation declined with developmental song learning. Nocturnal changes in HVC activity strongly and inversely correlated with the next days change, suggesting that sleep‐dependent degradation of HVC activity may facilitate or enable subsequent diurnal changes. Collectively, these data show that HVC activity levels exhibit daily cycles in adults and juveniles, whereas HVC burstiness and song stereotypy change daily in juveniles only. In addition, the data indicate that HVC burstiness increases with development and inversely correlates with song variability, which is necessary for trial and error vocal learning.

Identification of single neurons in a forebrain network

Behaviors are generated from complex interactions among networks of neurons. Single-unit ensemble recording has been used to identify multiple neurons in functioning networks. These recordings have provided insight into interactions among neurons in local and distributed circuits. Recorded units in these ensembles have been classed based on waveform type, firing pattern, and physical location. To identify individual projection neurons in a cortical network, we have paired tetrode recording with antidromic stimulation. We developed techniques that enable antidromic identification of single units and study of functional interactions between these neurons and other circuit elements. These methods have been developed in the zebra finch and should be applicable, with potential modifications that we discuss here, to any neural circuit with defined subpopulations based on projection target. This methodology will enable elucidation of the functional roles of single identified neurons in complex vertebrate circuits.

Models of vocal learning in the songbird: Historical frameworks and the stabilizing critic

Birdsong is a form of sensorimotor learning that involves a mirror‐like system that activates with both song hearing and production. Early models of song learning, based on behavioral measures, identified key features of vocal plasticity, such as the requirements for memorization of a tutor song and auditory feedback during song practice. The concept of a comparator, which compares the memory of the tutor song to auditory feedback, featured prominently. Later models focused on linking anatomically‐defined neural modules to behavioral concepts, such as the comparator. Exploiting the anatomical modularity of the songbird brain, localized lesions illuminated mechanisms of the neural song system. More recent models have integrated neuronal mechanisms identified in other systems with observations in songbirds. While these models explain multiple aspects of song learning, they must incorporate computational elements based on unknown biological mechanisms to bridge the motor‐to‐sensory delay and/or transform motor signals into the sensory domain. Here, I introduce the stabilizing critic hypothesis, which enables sensorimotor learning by (1) placing a purely sensory comparator afferent of the song system and (2) endowing song system disinhibitory interneuron networks with the capacity both to bridge the motor‐sensory delay through prolonged bursting and to stabilize song segments selectively based on the comparator signal. These proposed networks stabilize an otherwise variable signal generated by both putative mirror neurons and a cortical‐basal ganglia‐thalamic loop. This stabilized signal then temporally converges with a matched premotor signal in the efferent song motor cortex, promoting spike‐timing‐dependent plasticity in the premotor circuitry and behavioral song learning.

Rhythmic Cortical Neurons Increase their Oscillations and Sculpt Basal Ganglia Signaling During Motor Learning

The function and modulation of neural circuits underlying motor skill may involve rhythmic oscillations (Feller, ; Marder and Goaillard, ; Churchland et al., ). In the proposed pattern generator for birdsong, the cortical nucleus HVC, the frequency and power of oscillatory bursting during singing increases with development (Crandall et al., ; Day et al., ). We examined the maturation of cellular activity patterns that underlie these changes. Single unit ensemble recording combined with antidromic identification (Day et al., ) was used to study network development in anesthetized zebra finches. Autocovariance quantified oscillations within single units. A subset of neurons oscillated in the theta/alpha/mu/beta range (8–20 Hz), with greater power in adults compared to juveniles. Across the network, the normalized oscillatory power in the 8–20 Hz range was greater in adults than juveniles. In addition, the correlated activity between rhythmic neuron pairs increased with development. We next examined the functional impact of the oscillators on the output neurons of HVC. We found that the firing of oscillatory neurons negatively correlated with the activity of cortico‐basal ganglia neurons (HVCXs), which project to Area X (the song basal ganglia). If groups of oscillators work together to tonically inhibit and precisely control the spike timing of adult HVCXs with coordinated release from inhibition, then the activity of HVCXs in juveniles should be decreased relative to adults due to uncorrelated, tonic inhibition. Consistent with this hypothesis, HVCXs had lower activity in juveniles. These data reveal network changes that shape cortical‐to‐basal ganglia signaling during motor learning.