Todd F. Roberts

Rapid spine stabilization and synaptic enhancement at the onset of behavioural learning

Behavioural learning depends on the brain’s capacity to respond to instructive experience and is often enhanced during a juvenile sensitive period. How instructive experience acts on the juvenile brain to trigger behavioural learning remains unknown. In vitro studies show that forms of synaptic strengthening thought to underlie learning are accompanied by an increase in the stability, number and size of dendritic spines, which are the major sites of excitatory synaptic transmission in the vertebrate brain. In vivo imaging studies in sensory cortical regions reveal that these structural features can be affected by disrupting sensory experience and that spine turnover increases during sensitive periods for sensory map formation. These observations support two hypotheses: first, the increased capacity for behavioural learning during a sensitive period is associated with enhanced spine dynamics on sensorimotor neurons important for the learned behaviour; second, instructive experience rapidly stabilizes and strengthens these dynamic spines. Here we report a test of these hypotheses using two-photon in vivo imaging to measure spine dynamics in zebra finches, which learn to sing by imitating a tutor song during a juvenile sensitive period. Spine dynamics were measured in the forebrain nucleus HVC, the proximal site where auditory information merges with an explicit song motor representation, immediately before and after juvenile finches first experienced tutor song. Higher levels of spine turnover before tutoring correlated with a greater capacity for subsequent song imitation. In juveniles with high levels of spine turnover, hearing a tutor song led to the rapid (∼24-h) stabilization, accumulation and enlargement of dendritic spines in HVC. Moreover, in vivo intracellular recordings made immediately before and after the first day of tutoring revealed robust enhancement of synaptic activity in HVC. These findings suggest that behavioural learning results when instructive experience is able to rapidly stabilize and strengthen synapses on sensorimotor neurons important for the control of the learned behaviour.

A synaptic basis for auditory-vocal integration in the songbird

Songbirds learn to sing by memorizing a tutor song that they then vocally mimic using auditory feedback. This developmental sequence suggests that brain areas that encode auditory memories communicate with brain areas for learned vocal control. In the songbird, the secondary auditory telencephalic region caudal mesopallium (CM) contains neurons that encode aspects of auditory experience. We investigated whether CM is an important source of auditory input to two sensorimotor structures implicated in singing, the telencephalic song nucleus interface (NIf) and HVC. We used reversible inactivation methods to show that activity in CM is necessary for much of the auditory-evoked activity that can be detected in NIf and HVC of anesthetized adult male zebra finches. Furthermore, extracellular and intracellular recordings along with spike-triggered averaging methods indicate that auditory selectivity for the birds own song is enhanced between CM and NIf. We used lentiviral-mediated tracing methods to confirm that CM neurons directly innervate NIf. To our surprise, these tracing studies also revealed a direct projection from CM to HVC. We combined irreversible lesions of NIf with reversible inactivation of CM to establish that CM supplies a direct source of auditory drive to HVC. Finally, using chronic recording methods, we found that CM neurons are active in response to song playback and during singing, indicating their potential importance to song perception and processing of auditory feedback. These results establish the functional synaptic linkage between sites of auditory and vocal learning and may identify an important substrate for learned vocal communication.

Motor circuits are required to encode a sensory model for imitative learning

Premotor circuits help generate imitative behaviors and can be activated during observation of another animal′s behavior, leading to speculation that these circuits participate in sensory learning that is important to imitation. Here we tested this idea by focally manipulating the brain activity of juvenile zebra finches, which learn to sing by memorizing and vocally copying the song of an adult tutor. Tutor song–contingent optogenetic or electrical disruption of neural activity in the pupil′s song premotor nucleus HVC prevented song copying, indicating that a premotor structure important to the temporal control of birdsong also helps encode the tutor song. In vivo multiphoton imaging and neural manipulations delineated a pathway and a candidate synaptic mechanism through which tutor song information is encoded by premotor circuits. These findings provide evidence that premotor circuits help encode sensory information about the behavioral model before shaping and executing imitative behaviors.

Telencephalic Neurons Monosynaptically Link Brainstem and Forebrain Premotor Networks Necessary for Song

Birdsong, like human speech, is a series of learned vocal gestures resulting from the coordination of vocal and respiratory brainstem networks under the control of the telencephalon. The song motor circuit includes premotor and motor cortical analogs, known as HVC (used as a proper name) and RA (the robust nucleus of the arcopallium), respectively. Previous studies showed that HVC projects to RA and that RA projection neurons (PNs) topographically innervate brainstem vocal-motor and respiratory networks. The idea that singing-related activity flows between HVC and RA in a strictly feedforward manner is a central component of all models of song production. In contrast to this prevailing view of song motor circuit organization, we show that RA sends a reciprocal projection directly to HVC. Lentiviral labeling of RA PN axons and transgene tagging of RA PN synaptic terminals reveal a direct projection from RA to HVC. Retrograde tracing from HVC demonstrates that this projection originates exclusively from neurons in dorsocaudal regions of RA. Using dual retrograde tracer injections, we further show that many of these RAHVC neurons also innervate the brainstem nucleus retroambigualis, which is premotor to expiratory motoneurons, thereby identifying a population of RA PNs positioned to coordinate activity at higher and lower levels of the song motor circuit. In combination, our findings identify a previously unknown pathway that may enable a subset of RA neurons to provide song-related signals to the respiratory brainstem but also transmit a copy of this information to song patterning networks in HVC.

Organization of the avian basal forebrain: Chemical anatomy in the parrot (Melopsittacus undulatus)

Hodological, electrophysiological, and ablation studies indicate a role for the basal forebrain in telencephalic vocal control; however, to date the organization of the basal forebrain has not been extensively studied in any nonmammal or nonhuman vocal learning species. To this end the chemical anatomy of the avian basal forebrain was investigated in a vocal learning parrot, the budgerigar (Melopsittacus undulatus). Immunological and histological stains, including choline acetyltransferase, acetylcholinesterase, tyrosine hydroxylase, dopamine and cAMP‐regulated phosphoprotein (DARPP)‐32, the calcium binding proteins calbindin D‐28k and parvalbumin, calcitonin gene‐related peptide, iron, substance P, methionine enkephalin, nicotinamide adenine dinucleotide phosphotase diaphorase, and arginine vasotocin were used in the present study. We conclude that the ventral paleostriatum (cf. Kitt and Brauth [ 1981 ] Neuroscience 6:1551–1566) and adjacent archistriatal regions can be subdivided into several distinct subareas that are chemically comparable to mammalian basal forebrain structures. The nucleus accumbens is histochemically separable into core and shell regions. The nucleus taeniae (TN) is theorized to be homologous to the medial amygdaloid nucleus. The archistriatum pars ventrolateralis (Avl; comparable to the pigeon archistriatum pars dorsalis) is theorized to be a possible homologue of the central amygdaloid nucleus. The TN and Avl are histochemically continuous with the medial aspects of the bed nucleus of the stria terminalis and the ventromedial striatum, forming an avian analogue of the extended amygdala. The apparent counterpart in budgerigars of the mammalian nucleus basalis of Meynert consists of a field of cholinergic neurons spanning the basal forebrain. The budgerigar septal region is theorized to be homologous as a field to the mammalian septum. Our results are discussed with regard to both the evolution of the basal forebrain and its role in vocal learning processes. J. Comp. Neurol. 454:383–408, 2002.

Insights into the Neural and Genetic Basis of Vocal Communication

The use of vocalizations to communicate information and elaborate social bonds is an adaptation seen in many vertebrate species. Human speech is an extreme version of this pervasive form of communication. Unlike the vocalizations exhibited by the majority of land vertebrates, speech is a learned behavior requiring early sensory exposure and auditory feedback for its development and maintenance. Studies in humans and a small number of other species have provided insights into the neural and genetic basis for learned vocal communication and are helping to delineate the roles of brain circuits across the cortex, basal ganglia, and cerebellum in generating vocal behaviors. This Review provides an outline of the current knowledge about these circuits and the genes implicated in vocal communication, as well as a perspective on future research directions in this field.

Projections of the oval nucleus of the hyperstriatum ventrale in the budgerigar: Relationships with the auditory system

The afferent and efferent projections of a vocal control nucleus, the oval nucleus of the hyperstriatum ventrale (HVo), were mapped out in a parrot, the budgerigar (Melopsittacus undulatus) to determine the relationships of this nucleus to the auditory system. In budgerigars, HVo is connected to both the anterior forebrain pathway as well as to nuclei forming the descending projection system to the brainstem (Durand et al. [ 1997 ] J. Comp. Neurol. 377:179–206). Previous studies (Brauth et al. [ 1997 ] Proc. N. Y. Acad. Sci. 807:368–385; Durand and Brauth [ 1998 ] Neurosci Abstr 24:78.9) indicate that HVo lesions disrupt vocal performance and that HVo neurons show long latency electrophysiologic auditory responses. HVo has also been shown to receive input from neurons in the immediately adjacent HV (Durand et al. [ 1997 ] J. Comp. Neurol. 377:179–206). Thus, the focus of the present study was to elucidate relationships between HVo, its immediately adjacent surround and telencephalic auditory nuclei. The results show that, although the lateral and medial portions of HVo are interconnected with one another, inputs to these areas and their surrounds are distinctively different. The most substantial auditory system inputs are derived from the frontal lateral neostriatum (NFl) and supracentral nucleus of the lateral neostriatum (NLs); these project primarily to the lateral HVo and lateral HVo surround. The medial HVo and surround receive only sparse or modest input from auditory nuclei, including the caudomedial neostriatum (NCM), neostriatum intermedium pars lateralis (NIL), Fields L1 and L3, and the neostriatum intermedium pars ventrolateralis (NIVL). Other sources of input to the HVo surround include the hyperstriatum accessorium (HA), the supralaminar area of the frontal neostriatum (NAs), the ventral anterior archistriatum (AAv), the medial archistriatum (Am) and the medial HV. Neurons in the HV immediately medial to HVo project to a shell region around the entire nucleus. Both the ventral paleostriatum (VP) and ventral part of the central nucleus of the lateral neostriatum (NLc) project to HVo but not to the surround. Previously described projections (Durand et al., 1997 ) from HVo to the NAom, NLc, and the magnicellular nucleus of the lobus parolfactorius (LPOm) were confirmed. J. Comp. Neurol. 432:481–511, 2001.

Distribution of tyrosine hydroxylase-containing neurons and fibers in the brain of the budgerigar (Melopsittacus undulatus): general patterns and labeling in vocal control nuclei.

The distribution of tyrosine hydroxylase (TH) was mapped out in cells and fibers of the budgerigar (Melopsittacus undulatus) brain. Special attention was given to vocal control and auditory nuclei because budgerigars are a psittacine species in which both males and females are capable of lifelong vocal learning (Farabaugh et al. [1994] J. Comp. Psychol 108:81–92). The results show that TH staining in the central nucleus of the anterior archistriatum (AAc) resembled that of surrounding archistriatal fields, except for portions of the ventral archistriatum, which exhibited substantially more TH+ fibers. Fewer fibers and fiber baskets are present in the central nucleus of the lateral neostriatum (NLc) than in surrounding fields. Both the oval nuclei of the ventral hyperstriatum (HVo) and anterior neostriatum (NAo) exhibit less fiber staining than surrounding fields whereas fiber staining in the medial NAo (NAom) and magnicellular nucleus of the parolfactory lobe (LPOm) resemble that of surrounding fields. Staining in primary telencephalic auditory nuclei was extremely low. The only sex difference observed was slightly increased TH staining in LPOm of females compared with surrounding fields on some tissue sections. These findings are in contrast to previous findings in zebra finch (Poephila guttata), a close ended vocal learning songbird in which TH staining in vocal nuclei increases during development and remains greater than surrounding fields throughout adulthood. The present results therefore support the view that catecholamines act to inhibit vocal plasticity in adult vocal learning species. Several unique features of TH‐immunoreactive (ir) cell groups were observed in the brainstem including sparsely scattered TH‐ir somata immediately adjacent to the third ventricle, within the tectum, basal forebrain, archistriatum, and caudal neostriatum, and in the hippocampus. These latter populations have not been described in other avian species and resemble features of the catecholamine system generally found in either reptiles or mammals. J. Comp. Neurol. 429:436–454, 2001.

Animal Models of Speech and Vocal Communication Deficits Associated With Psychiatric Disorders

Disruptions in speech, language, and vocal communication are hallmarks of several neuropsychiatric disorders, most notably autism spectrum disorders. Historically, the use of animal models to dissect molecular pathways and connect them to behavioral endophenotypes in cognitive disorders has proven to be an effective approach for developing and testing disease-relevant therapeutics. The unique aspects of human language compared with vocal behaviors in other animals make such an approach potentially more challenging. However, the study of vocal learning in species with analogous brain circuits to humans may provide entry points for understanding this human-specific phenotype and diseases. We review animal models of vocal learning and vocal communication and specifically link phenotypes of psychiatric disorders to relevant model systems. Evolutionary constraints in the organization of neural circuits and synaptic plasticity result in similarities in the brain mechanisms for vocal learning and vocal communication. Comparative approaches and careful consideration of the behavioral limitations among different animal models can provide critical avenues for dissecting the molecular pathways underlying cognitive disorders that disrupt speech, language, and vocal communication.

3.23 – Neurophysiology of Birdsong Learning

Oscine songbirds use auditory feedback to learn and, in some species, to maintain their courtship songs. Song learning is restricted to a juvenile sensitive period characterized by a remarkable capacity for memorization and subsequent accurate imitation of tutor songs. The songbird’s brain contains a constellation of interconnected brain nuclei, known as the song system, which plays an important role in singing and song learning. This chapter covers our current understanding of the ecological function of song, the peripheral and central mechanisms of song production and the neural mechanisms of song learning.