Vocal Learning In Songbirds Research Articles

Auditory Feedback and The Plasticity of Vocal Learning in Songbirds*

Songbirds and humans both rely critically on auditory feedback for learning and maintaining accurate vocalizations. During vocal learning, juvenile songbirds learn to sing by using auditory feedback to match their variable song patterns to a memorized tutor model. During song maintaining, adult songbirds monitor the integrality and accuracy of its song on-line according to auditory feedback. This form of learning requires auditory-vocal integration. The output of songs and the information of auditory feedback are integrated in some areas of the bird's brain, and then direct next vocal activity. In recent years, the lateral magnocellular nucleus of the anterior neostriatum (LMAN) has been demonstrated that it is necessary for feedback-dependent song decrystallization, and LMAN neurons exhibit highly selective responses to auditory presentation of the bird's own song (BOS). Moreover, the discovery of mirror neurons in the high vocal centre (HVC) also provides a significant clue for further study in songbirds. Here the progress of auditory feedback and the plasticity of vocal learning in songbirds are reviewed in recent years.

Activity Propagation in an Avian Basal Ganglia-Thalamocortical Circuit Essential for Vocal Learning

In mammalian basal ganglia-thalamocortical circuits, GABAergic pallidal neurons are thought to "gate" or modulate excitation in thalamus with their strong inhibitory inputs and thus signal to cortex by pausing and permitting thalamic neurons to fire in response to excitatory drive. In contrast, in a homologous circuit specialized for vocal learning in songbirds, evidence suggests that pallidal neurons signal by eliciting postinhibitory rebound spikes in thalamus, which could occur even without any excitatory drive to thalamic neurons. To test whether songbird pallidal neurons can also communicate with thalamus by gating excitatory drive, as well as by postinhibitory rebound, we examined the activity of thalamic relay neurons in response to acute inactivation of the basal ganglia structure Area X; Area X contains the pallidal neurons that project to thalamus. Although inactivation of Area X should eliminate rebound-mediated spiking in thalamus, this manipulation tonically increased the firing rate of thalamic relay neurons, providing evidence that songbird pallidal neurons can gate tonic thalamic excitatory drive. We also found that the increased thalamic activity was fed forward to its target in the avian equivalent of cortex, which includes neurons that project to the vocal premotor area. These data raise the possibility that basal ganglia circuits can signal to cortex through thalamus both by generating postinhibitory rebound and by gating excitatory drive and may switch between these modes depending on the statistics of pallidal firing. Moreover, these findings provide insight into the strikingly different disruptive effects of basal ganglia and cortical lesions on songbird vocal learning.

Top-Down Regulation of Plasticity in the Birdsong System: “Premotor” Activity in the Nucleus HVC Predicts Song Variability Better Than It Predicts Song Features

We studied real-time changes in brain activity during active vocal learning in the zebra finch songbird. The song nucleus HVC is required for the production of learned song. To quantify the relationship of HVC activity and behavior, HVC population activity during repeated vocal sequences (motifs) was recorded and temporally aligned relative to the motif, millisecond by millisecond. Somewhat surprisingly, HVC activity did not reliably predict any vocal feature except amplitude and, to a lesser extent, entropy and pitch goodness (sound periodicity). Variance in "premotor" HVC activity did not reliably predict variance in behavior. In contrast, HVC activity inversely predicted the variance of amplitude, entropy, frequency, pitch, and FM. We reasoned that, if HVC was involved in song learning, the relationship of HVC activity to learned features would be developmentally regulated. To test this hypothesis, we compared the HVC song feature relationships in adults and juveniles in the sensorimotor "babbling" period. We found that the relationship of HVC activity to variance in FM was developmentally regulated, with the greatest difference at an HVC vocalization lag of 50 ms. Collectively, these data show that, millisecond by millisecond, bursts in HVC activity predict song stability on-line during singing, whereas decrements in HVC activity predict plasticity. These relationships between neural activity and plasticity may play a role in vocal learning in songbirds by enabling the selective stabilization of parts of the song that match a learned tutor model.

GABA immunoreactivity in auditory and song control brain areas of zebra finches

Inhibitory transmission is critical to sensory and motor processing and is believed to play a role in experience-dependent plasticity. The main inhibitory neurotransmitter in vertebrates, GABA, has been implicated in both sensory and motor aspects of vocalizations in songbirds. To understand the role of GABAergic mechanisms in vocal communication, GABAergic elements must be characterized fully. Hence, we investigated GABA immunohistochemistry in the zebra finch brain, emphasizing auditory areas and song control nuclei. Several nuclei of the ascending auditory pathway showed a moderate to high density of GABAergic neurons including the cochlear nuclei, nucleus laminaris, superior olivary nucleus, mesencephalic nucleus lateralis pars dorsalis, and nucleus ovoidalis. Telencephalic auditory areas, including field L subfields L1, L2a and L3, as well as the caudomedial nidopallium (NCM) and mesopallium (CMM), contained GABAergic cells at particularly high densities. Considerable GABA labeling was also seen in the shelf area of caudodorsal nidopallium, and the cup area in the arcopallium, as well as in area X, the lateral magnocellular nucleus of the anterior nidopallium, the robust nucleus of the arcopallium and nidopallial nucleus HVC. GABAergic cells were typically small, most likely local inhibitory interneurons, although large GABA-positive cells that were sparsely distributed were also identified. GABA-positive neurites and puncta were identified in most nuclei of the ascending auditory pathway and in song control nuclei. Our data are in accordance with a prominent role of GABAergic mechanisms in regulating the neural circuits involved in song perceptual processing, motor production, and vocal learning in songbirds.

Developmental origin and identity of song system neurons born during vocal learning in songbirds

New neurons are added to the forebrain song control regions high vocal center (HVC) and Area X of juvenile songbirds but the identity and site of origin of these cells have not been fully characterized. We used oncoretroviral vectors to genetically label neuronal progenitors in different regions of the zebra finch lateral ventricle. A region corresponding to the mammalian medial and lateral ganglionic eminences generated medium spiny neurons found in Area X and in the striatum surrounding Area X, and at least two classes of interneurons found in HVC. In addition, our experiments indicate that the HVC projection neurons that project into nucleus robust nucleus of the arcopallium (RA) are born locally from the ventricular region immediately dorsal to HVC. The ability to genetically target neuron subpopulations that give rise to different song system cell types provides a tool for specific genetic manipulations of these cell types. In addition, our results suggest striking similarities between neurogenesis in the embryonic mammalian brain and in the brain of the juvenile songbird and provide further evidence for the existence of conserved cell types in the forebrain for birds and mammals.

Isolation of song nuclei specific genes by using cDNA microarray in Bengalese finch

Inhibitory transmission is critical to sensory and motor processing and is believed to play a role in experience-dependent plasticity. The main inhibitory neurotransmitter in vertebrates, GABA, has been implicated in both sensory and motor aspects of vocalizations in songbirds. To understand the role of GABAergic mechanisms in vocal communication, GABAergic elements must be characterized fully. Hence, we investigated GABA immunohistochemistry in the zebra finch brain, emphasizing auditory areas and song control nuclei. Several nuclei of the ascending auditory pathway showed a moderate to high density of GABAergic neurons including the cochlear nuclei, nucleus laminaris, superior olivary nucleus, mesencephalic nucleus lateralis pars dorsalis, and nucleus ovoidalis. Telencephalic auditory areas, including field L subfields L1, L2a and L3, as well as the caudomedial nidopallium (NCM) and mesopallium (CMM), contained GABAergic cells at particularly high densities. Considerable GABA labeling was also seen in the shelf area of caudodorsal nidopallium, and the cup area in the arcopallium, as well as in area X, the lateral magnocellular nucleus of the anterior nidopallium, the robust nucleus of the arcopallium and nidopallial nucleus HVC. GABAergic cells were typically small, most likely local inhibitory interneurons, although large GABA-positive cells that were sparsely distributed were also identified. GABA-positive neurites and puncta were identified in most nuclei of the ascending auditory pathway and in song control nuclei. Our data are in accordance with a prominent role of GABAergic mechanisms in regulating the neural circuits involved in song perceptual processing, motor production, and vocal learning in songbirds.

Early auditory experience generates long-lasting memories that may subserve vocal learning in songbirds.

In both humans and songbirds, infants learn vocalizations by imitating the sounds of adult tutors with whom they interact during an early sensitive period. Vocal learning occurs in few animal taxa; similarities in the imitation process between humans and songbirds make the songbird a unique system in which vocal learning mechanisms can be studied at the neurobiological level. One theory of vocal learning proposes that early auditory experience generates auditory memories that subsequently guide vocal imitation. We now present a combination of behavioral and neurophysiological results, obtained in a songbird, that support this theory. We show that neurons in a forebrain auditory area of adult male zebra finches are selectively tuned to the song of a tutor heard early in development. Furthermore, the strength of this selectivity shows a striking correlation with the fidelity of vocal imitation, suggesting that this auditory memory may have served as the model for song learning.

Silent Synapses in a Thalamo-Cortical Circuit Necessary for Song Learning in Zebra Finches

Developmental changes in synaptic properties may act to limit neural and behavioral plasticity associated with sensitive periods. This study characterized synaptic maturation in a glutamatergic thalamo-cortical pathway that is necessary for vocal learning in songbirds. Lesions of the projection from medial dorsolateral nucleus of the thalamus (DLM) to the cortical nucleus lateral magnocellular nucleus of the anterior nidopallium (LMAN) greatly disrupt song behavior in juvenile birds during early stages of vocal learning. However, such lesions lose the ability to disrupt vocal behavior in normal birds at 60-70 days of age, around the time that selective auditory tuning for each bird's own song (BOS) emerges in LMAN neurons. This pattern has suggested that LMAN is involved in processing song-related information and evaluating the degree to which vocal motor output matches the tutor song to be learned. Analysis of reversed excitatory postsynaptic currents at DLM-->LMAN synapses in in vitro slice preparations revealed a pronounced N-methyl-D-aspartate receptor (NMDAR)-mediated component in both juvenile and adult cells with no developmental decrease in the relative contribution of NMDARs to synaptic transmission. However, the synaptic failure rate at DLM-->LMAN synapses in juvenile males during the sensitive period for song learning was significantly lower at depolarized potentials than at hyperpolarized potentials. In contrast, the failure rate at DLM-->LMAN synapses did not differ at hyper- versus depolarized holding potentials in adult males that had completed the acquisition of a stereotyped song. This pattern indicates that juvenile cells have a higher incidence of silent (NMDAR-only) synapses, which are postsynaptically silent at hyperpolarized potentials due to the voltage-dependent gating of NMDARs. Thus the decreased involvement of the LMAN pathway in vocal behavior is mirrored by a decline in the incidence of silent synapses but not by changes in the relative number of NMDA and AMPA receptors at DLM-->LMAN synapses. These findings suggest that a developmental decrease in silent synapses within LMAN may represent a neural correlate of behavioral plasticity during song learning.

Timing and Prediction

When is an inhibitory synapse not inhibitory? In this issue of Neuron, Person and Perkel demonstrate that thalamic neurons can translate extrinsic GABAergic input from the basal ganglia into highly precise patterns of sustained spiking in a circuit that is essential for vocal learning in songbirds. Postinhibitory rebound serves as a mechanism that preserves precise spike timing information, enabling reliable propagation of activity throughout this pathway. The results have broad implications for basic mechanisms of functional processing in both thalamus and basal ganglia and serve to increase our understanding of how acoustic units of vocal sounds are transformed into motor gestures during the sensitive period for song learning.

Properties of Dopamine Release and Uptake in the Songbird Basal Ganglia

Vocal learning in songbirds requires a basal ganglia circuit termed the anterior forebrain pathway (AFP). The AFP is not required for song production, and its role in song learning is not well understood. Like the mammalian striatum, the striatal component of the AFP, Area X, receives dense dopaminergic innervation from the midbrain. Since dopamine (DA) clearly plays a crucial role in basal ganglia-mediated motor control and learning in mammals, it seems likely that DA signaling contributes importantly to the functions of Area X as well. In this study, we used voltammetric methods to detect subsecond changes in extracellular DA concentration to gain better understanding of the properties and regulation of DA release and uptake in Area X. We electrically stimulated Ca(2+)- and action potential-dependent release of an electroactive substance in Area X brain slices and identified the substance as DA by the voltammetric waveform, electrode selectivity, and neurochemical and pharmacological evidence. As in the mammalian striatum, DA release in Area X is depressed by autoinhibition, and the lifetime of extracellular DA is strongly constrained by monoamine transporters. These results add to the known physiological similarities of the mammalian and songbird striatum and support further use of voltammetry in songbirds to investigate the role of basal ganglia DA in motor learning.

Developmental regulation of basal ganglia circuitry during the sensitive period for vocal learning in songbirds.

A hallmark of sensitive periods of development is an enhanced capacity for learning, such that experience exerts a profound effect on the brain resulting in the establishment of behaviors and underlying neural circuitry that can last a lifetime. Songbirds, like humans, have a sensitive period for vocal learning: they acquire the sounds used for vocal communication during a restricted period of development. In principle, any organism that undertakes vocal learning is faced with the same challenge: to form some representation of target vocal sounds based on auditory experience, and then to translate that auditory target into a motor program that reproduces the sound. Both birds and humans achieve this translation by using auditory (and other) feedback resulting from incipient vocalizations ("babbling" in humans, "subsong" in birds) to adjust motor commands until vocal output produces a good copy of the target sounds. Similarities between vocal learning in birds and humans suggest that many aspects of the learning process have evolved to meet demands imposed by vocal communication. Thus songbirds provide a valuable animal model in which to study the physiological basis of learned vocal communication and the nature of sensitive periods in general. In this article, I describe aspects of both behavioral and neural frameworks that currently inform our thinking about mechanisms underlying vocal learning and behavior in songbirds, and highlight ideas that may need re-examination.

Long-term potentiation in an avian basal ganglia nucleus essential for vocal learning.

Vocal learning in songbirds provides an excellent model for sensorimotor learning in vertebrates, with an accessible, well-defined behavior and discrete neural substrate. The rich behavioral plasticity exhibited by songbirds, however, contrasts starkly with the scarcity of candidate cellular mechanisms. Here, we report for the first time on an activity-dependent form of synaptic plasticity in area X, a component of the song system required for song learning and song maintenance. In slice preparations of zebra finch area X, pairing of high-frequency presynaptic stimulation with postsynaptic depolarization induces Hebbian long-term potentiation (LTP) of the glutamatergic inputs to spiny neurons. This form of LTP requires activation of NMDA receptors and D1-like dopamine receptors. In addition, LTP is observed in birds as young as 47 d after hatching and also in adult birds but not in younger birds, providing evidence of developmental regulation of the onset of synaptic plasticity. These properties make this form of LTP the best known candidate mechanism for reinforcement-based vocal learning in juveniles and song maintenance in adult birds.

Gene regulation by song in the auditory telencephalon of songbirds.

Inducible gene expression analysis has been successfully used to identify and characterize areas involved in the auditory processing of song in songbirds. When songbirds hear song, zenk, a gene encoding a transcriptional regulator linked to synaptic plasticity, is rapidly and transiently induced in several discrete areas of the telencephalon. This phenomenon is most marked in the caudomedial neostriatum (NCM), a major auditory processing area. zenk induction by song has proven very useful to investigate the functional organization of NCM as well as to characterize song-responding neurons in this brain area. Combined with data from anatomical and electrophysiological studies, the results of gene expression analysis suggest that NCM is involved in the auditory processing of complex sounds such as song and possibly in the formation and/or storage of song auditory memories. Thus, NCM appears to play a prominent role in perceptual aspects of vocal communication, and potentially in the process of vocal learning in songbirds and other vocal learning avian orders, i.e. parrots and hummingbirds.

Vocal learning in birds and humans.

Vocal learning is the modification of vocal output by reference to auditory information. It allows for the imitation and improvisation of sounds that otherwise would not occur. The emergence of this skill may have been a primary step in the evolution of human language, but vocal learning is not unique to humans. It also occurs in songbirds, where its biology can be studied with greater ease. What follows is a review of some of the salient anatomical, developmental, and behavioral features of vocal learning, alongside parallels and differences between vocal learning in songbirds and humans.

Dopamine Modulates Excitability of Spiny Neurons in the Avian Basal Ganglia

The neural substrate of vocal learning in songbirds is an accessible system for studying motor learning and motor control in vertebrates. In the so-called song system, the anterior forebrain pathway (AFP), which is essential for song learning, resembles the mammalian basal ganglia-thalamocortical loop in its macroscopic organization, neuronal intrinsic properties, and microcircuitry. Area X, the first station in the AFP, and the surrounding lobus parolfactorius (LPO), are both parts of the avian basal ganglia. Like their mammalian counterparts, they receive dense dopaminergic innervation from the midbrain, but the physiological functions of this projection remain unclear. In this study, we investigated the effect of dopamine (DA) on excitability of spiny neurons in area X and LPO. We recorded from neurons in brain slices of adult zebra finches and Bengalese finches, using whole-cell and perforated-patch recording techniques in current-clamp configuration. We found that DA modulates excitability in spiny neurons; activation of D1- and D2-like DA receptors enhances and reduces excitability, respectively. These effects are similar to those observed in the mammalian neostriatum, with the main difference being that D1-like DA receptor activation enhances excitability in avian spiny neurons at hyperpolarized states. Our findings also indicate that some spiny neurons express both receptor types and suggest that receptor colocalization in the entire population can account for the spectrum of DA actions. The diversity of DA actions enables the DA system to fine-tune the dynamics of the song system and allows flexible control over song learning and production.

Increased expression of endogenous biotin, but not BDNF, in telencephalic song regions during zebra finch vocal learning

Brain-derived neurotrophic factor (BDNF) is thought to regulate multiple aspects of brain development and neural plasticity in vertebrates. We have examined BDNF expression in two telencephalic nuclei (RA and HVC) in the zebra finch brain that control song learning by juvenile males and the production of already-learned song by adults. Using two different antibody-labeling techniques (avidin–biotin complex and horseradish peroxidase), we were unable to detect BDNF-like immunoreactivity in RA of juvenile or adult birds. BDNF-like immunoreactive labeling of somata was detected in HVC, but the density of labeled cells was not different between juvenile and adult birds. Immunocytochemical findings were confirmed by RT-PCR for BDNF mRNA. Thus, in contrast to a previous report (Akutagawa and Konishi, Proc. Natl. Acad. Sci. USA 95 (1998) 11429–11434), we did not observe elevated levels of BDNF immunoreactivity in RA and HVC of juvenile birds that were learning to sing. However, RA and HVC of juvenile birds were found to express elevated levels of endogenous biotin (as detected by avidin peroxidase), suggesting a possible role for biotin-regulated mechanisms in songbird vocal learning.

The neural basis of vocal learning in songbirds

The neural basis of vocal learning in songbirds

Intrinsic and synaptic properties of neurons in the vocal‐control nucleus lMAN from in vitro slice preparations of juvenile and adult zebra finches

A common theme of diverse neural systems is that circuits that are important for initial acquisition of learning do not necessarily serve as a substrate for the long-term storage of that memory. The neural basis of vocal learning in songbirds provides an example of this phenomenon, since a circuit that is necessary for vocal production during initial stages of vocal development apparently plays no subsequent role in controlling learned vocalizations. This striking functional change suggests the possibility of marked physiological changes in synaptic transmission within this circuit. We therefore examined intrinsic and synaptic properties of neurons in the cortical nucleus lMAN (lateral magnocellular nucleus of the anterior neostriatum), which forms part of this developmentally regulated circuit, in an in vitro preparation of the zebra finch forebrain. Although both functional and morphological characteristics of these neurons change substantially during vocal development, we did not observe widespread, substantive changes in the electrophysiological characteristics of juvenile versus adult lMAN neurons examined in vitro. Overall, both the intrinsic properties and synaptic responses of lMAN neurons were similar in slices from juvenile birds (at ages when lesions of lMAN disrupt vocal production) and in slices from adult birds (when lMAN lesions have no effect on song production). However, one intrinsic property that did vary between juvenile and adult cells was spike duration, which was longer in juvenile cells, suggesting the potential for activation of second-messenger cascades and/or enhanced synaptic transmission onto target cells of lMAN neurons. The pattern of synaptic response observed in both juvenile and adult cells suggests that lMAN projection neurons receive direct excitatory afferent inputs, as well as disynaptic inhibitory inputs from interneurons within lMAN. Activation of inhibitory interneurons rapidly curtails the excitatory response seen in projection neurons. This inhibition was abolished by bicuculline, indicating that the inhibitory interneurons normally exert their postsynaptic response via GABAA receptors on projection neurons. The inhibitory response could also be blocked by CNQX (6-cyano-7-nitroquinoxaline-2,3-dione), suggesting that the activation of inhibitory interneurons within lMAN may be governed primarily by AMPA receptors. © 1998 John Wiley & Sons, Inc. J Neurobiol 37: 642–658, 1998

Intrinsic and synaptic properties of neurons in the vocal-control nucleus lMAN fromin vitro slice preparations of juvenile and adult zebra finches

A common theme of diverse neural systems is that circuits that are important for initial acquisition of learning do not necessarily serve as a substrate for the long-term storage of that memory. The neural basis of vocal learning in songbirds provides an example of this phenomenon, since a circuit that is necessary for vocal production during initial stages of vocal development apparently plays no subsequent role in controlling learned vocalizations. This striking functional change suggests the possibility of marked physiological changes in synaptic transmission within this circuit. We therefore examined intrinsic and synaptic properties of neurons in the cortical nucleus IMAN (lateral magnocellular nucleus of the anterior neostriatum), which forms part of this developmentally regulated circuit, in an in vitro preparation of the zebra finch forebrain. Although both functional and morphological characteristics of these neurons change substantially during vocal development, we did not observe widespread, substantive changes in the electrophysiological characteristics of juvenile versus adult IMAN neurons examined in vitro. Overall, both the intrinsic properties and synaptic responses of IMAN neurons were similar in slices from juvenile birds (at ages when lesions of IMAN disrupt vocal production) and in slices from adult birds (when IMAN lesions have no effect on song production). However, one intrinsic property that did vary between juvenile and adult cells was spike duration, which was longer in juvenile cells, suggesting the potential for activation of second-messenger cascades and/or enhanced synaptic transmission onto target cells of IMAN neurons. The pattern of synaptic response observed in both juvenile and adult cells suggests that IMAN projection neurons receive direct excitatory afferent inputs, as well as disynaptic inhibitory inputs from interneurons within IMAN. Activation of inhibitory interneurons rapidly curtails the excitatory response seen in projection neurons. This inhibition was abolished by bicuculline, indicating that the inhibitory interneurons normally exert their postsynaptic response via GABA(A) receptors on projection neurons. The inhibitory response could also be blocked by CNQX (6-cyano-7-nitroquinoxaline-2,3-dione), suggesting that the activation of inhibitory interneurons within IMAN may be governed primarily by AMPA receptors.

Descending auditory pathways in the adult male zebra finch (Taeniopygia Guttata)

Here, we examine the connectivity of two previously identified telencephalic stations of the auditory system of adult zebra finches, the neostriatal "shelf" that underlies the high vocal center (HVC) and the archistriatal "cup" adjacent to the robust nucleus of the archistriatum (RA). We used different kinds of neuroanatomical tracers to visualize the projections from the shelf to the HVC. In addition, we show that the shelf projects to the cup and that the cup projects to thalamic, midbrain, and pontine nuclei of the ascending auditory pathway. Our observations extend to songbirds anatomical features that are found in the auditory pathways of a nonoscine bird, the pigeon (Wild et al. [1993] J. Comp. Neurol. 337:32-62), and we suggest that the descending auditory projections found in mammals may also be a general property of the avian brain. Finally, we show that the oscine song control system is closely apposed to auditory pathways at many levels. Our observations may help in understanding the evolution and organization of networks for vocal communication and vocal learning in songbirds.