Familiar But Unexpected: Effects of Sound Context Statistics on Auditory Responses in the Songbird Forebrain.

Rapid discrimination of salient acoustic signals in the noisy natural environment may depend, not only on specific stimulus features, but also on previous experience that generates expectations about upcoming events. We studied the neural correlates of expectation in the songbird forebrain by using natural vocalizations as stimuli and manipulating the category and familiarity of context sounds. In our paradigm, we recorded bilaterally from auditory neurons in awake adult male zebra finches with multiple microelectrodes during repeated playback of a conspecific song, followed by further playback of this test song in different interleaved sequences with other conspecific or heterospecific songs. Significant enhancement in the auditory response to the test song was seen when its acoustic features differed from the statistical distribution of context song features, but not when it shared the same distribution. Enhancement was also seen when the time of occurrence of the test song was uncertain. These results show that auditory forebrain responses in awake animals in the passive hearing state are modulated dynamically by previous auditory experience and imply that the auditory system can identify the category of a sound based on the global features of the acoustic context. Furthermore, this probability-dependent enhancement in responses to surprising stimuli is independent of stimulus-specific adaptation, which tracks familiarity, suggesting that the two processes could coexist in auditory processing. These findings establish the songbird as a model system for studying these phenomena and contribute to our understanding of statistical learning and the origin of human ERP phenomena to unexpected stimuli.SIGNIFICANCE STATEMENT Traditional auditory neurophysiology has mapped acoustic features of sounds to the response properties of neurons; however, growing evidence suggests that neurons can also encode the probability of sounds. We recorded responses of songbird auditory neurons in a novel paradigm that presented a familiar test stimulus in a sequence with similar or dissimilar sounds. The responses encode, not only stimulus familiarity, but also the expectation for a class of sounds based on the recent statistics of varying sounds in the acoustic context. Our approach thus provides a model system that uses a controlled stimulus paradigm to understand the mechanisms by which top-down processes (expectation and memory) and bottom-up processes (based on stimulus features) interact in sensory coding.

Principles of auditory processing differ between sensory and premotor structures of the songbird forebrain.

Sensory and motor brain structures work in collaboration during perception. To evaluate their respective contributions, the present study recorded neural responses to auditory stimulation at multiple sites simultaneously in both the higher-order auditory area NCM and the premotor area HVC of the songbird brain in awake zebra finches (Taeniopygia guttata). Bird's own song (BOS) and various conspecific songs (CON) were presented in both blocked and shuffled sequences. Neural responses showed plasticity in the form of stimulus-specific adaptation, with markedly different dynamics between the two structures. In NCM, the response decrease with repetition of each stimulus was gradual and long-lasting and did not differ between the stimuli or the stimulus presentation sequences. In contrast, HVC responses to CON stimuli decreased much more rapidly in the blocked than in the shuffled sequence. Furthermore, this decrease was more transient in HVC than in NCM, as shown by differential dynamics in the shuffled sequence. Responses to BOS in HVC decreased more gradually than to CON stimuli. The quality of neural representations, computed as the mutual information between stimuli and neural activity, was higher in NCM than in HVC. Conversely, internal functional correlations, estimated as the coherence between recording sites, were greater in HVC than in NCM. The cross-coherence between the two structures was weak and limited to low frequencies. These findings suggest that auditory communication signals are processed according to very different but complementary principles in NCM and HVC, a contrast that may inform study of the auditory and motor pathways for human speech processing.NEW & NOTEWORTHY Neural responses to auditory stimulation in sensory area NCM and premotor area HVC of the songbird forebrain show plasticity in the form of stimulus-specific adaptation with markedly different dynamics. These two structures also differ in stimulus representations and internal functional correlations. Accordingly, NCM seems to process the individually specific complex vocalizations of others based on prior familiarity, while HVC responses appear to be modulated by transitions and/or timing in the ongoing sequence of sounds.

Statistical learning of recurring sound patterns encodes auditory objects in songbird forebrain.

Auditory neurophysiology has demonstrated how basic acoustic features are mapped in the brain, but it is still not clear how multiple sound components are integrated over time and recognized as an object. We investigated the role of statistical learning in encoding the sequential features of complex sounds by recording neuronal responses bilaterally in the auditory forebrain of awake songbirds that were passively exposed to long sound streams. These streams contained sequential regularities, and were similar to streams used in human infants to demonstrate statistical learning for speech sounds. For stimulus patterns with contiguous transitions and with nonadjacent elements, single and multiunit responses reflected neuronal discrimination of the familiar patterns from novel patterns. In addition, discrimination of nonadjacent patterns was stronger in the right hemisphere than in the left, and may reflect an effect of top-down modulation that is lateralized. Responses to recurring patterns showed stimulus-specific adaptation, a sparsening of neural activity that may contribute to encoding invariants in the sound stream and that appears to increase coding efficiency for the familiar stimuli across the population of neurons recorded. As auditory information about the world must be received serially over time, recognition of complex auditory objects may depend on this type of mnemonic process to create and differentiate representations of recently heard sounds.

Neural responses in songbird forebrain reflect learning rates, acquired salience, and stimulus novelty after auditory discrimination training.

How do social interactions form and modulate the neural representations of specific complex signals? This question can be addressed in the songbird auditory system. Like humans, songbirds learn to vocalize by imitating tutors heard during development. These learned vocalizations are important in reproductive and social interactions and in individual recognition. As a model for the social reinforcement of particular songs, male zebra finches were trained to peck for a food reward in response to one song stimulus (GO) and to withhold responding for another (NoGO). After performance reached criterion, single and multiunit neural responses to both trained and novel stimuli were obtained from multiple electrodes inserted bilaterally into two songbird auditory processing areas [caudomedial mesopallium (CMM) and caudomedial nidopallium (NCM)] of awake, restrained birds. Neurons in these areas undergo stimulus-specific adaptation to repeated song stimuli, and responses to familiar stimuli adapt more slowly than to novel stimuli. The results show that auditory responses differed in NCM and CMM for trained (GO and NoGO) stimuli vs. novel song stimuli. When subjects were grouped by the number of training days required to reach criterion, fast learners showed larger neural responses and faster stimulus-specific adaptation to all stimuli than slow learners in both areas. Furthermore, responses in NCM of fast learners were more strongly left-lateralized than in slow learners. Thus auditory responses in these sensory areas not only encode stimulus familiarity, but also reflect behavioral reinforcement in our paradigm, and can potentially be modulated by social interactions.

Hemispheric differences in processing of vocalizations depend on early experience.

An intriguing phenomenon in the neurobiology of language is lateralization: the dominant role of one hemisphere in a particular function. Lateralization is not exclusive to language because lateral differences are observed in other sensory modalities, behaviors, and animal species. Despite much scientific attention, the function of lateralization, its possible dependence on experience, and the functional implications of such dependence have yet to be clearly determined. We have explored the role of early experience in the development of lateralized sensory processing in the brain, using the songbird model of vocal learning. By controlling exposure to natural vocalizations (through isolation, song tutoring, and muting), we manipulated the postnatal auditory environment of developing zebra finches, and then assessed effects on hemispheric specialization for communication sounds in adulthood. Using bilateral multielectrode recordings from a forebrain auditory area known to selectively process species-specific vocalizations, we found that auditory responses to species-typical songs and long calls, in both male and female birds, were stronger in the right hemisphere than in the left, and that right-side responses adapted more rapidly to stimulus repetition. We describe specific instances, particularly in males, where these lateral differences show an influence of auditory experience with song and/or the bird's own voice during development.

Dynamic encoding of phonetic categories in zebra finch auditory forebrain.

Vocal communication requires the formation of acoustic categories to enable invariant representations of sounds despite superficial variations. Humans form acoustic categories for speech phonemes, enabling the listener to recognize words independent of speakers; animals can also discriminate speech phonemes. We investigated the neural mechanisms of this process using electrophysiological recordings from the zebra finch secondary auditory area, caudomedial nidopallium (NCM), during passive exposure to human speech stimuli consisting of two naturally spoken words produced by multiple speakers. Analysis of neural distance and decoding accuracy showed improvements in neural discrimination between word categories over the course of exposure, and this improved representation transferred to the same words by novel speakers. We conclude that NCM neurons formed generalized representations of word categories independent of speaker-specific variations that became more refined over the course of passive exposure. The discovery of this dynamic encoding process in NCM suggests a general processing mechanism for forming categorical representations of complex acoustic signals that humans share with other animals.

Novel sound exposure drives dynamic changes in auditory lateralization that are associated with perceptual learning in zebra finches.

Songbirds provide a model for adult plasticity in the auditory cortex as a function of recent experience due to parallels with human auditory processing. As for speech processing in humans, activity in songbirds' higher auditory cortex (caudomedial nidopallium, NCM) is lateralized for complex vocalization sounds. However, in Zebra finches exposed to a novel heterospecific (canary) acoustic environment for 4-9 days, the typical pattern of right-lateralization is reversed. We now report that, in birds passively exposed to a novel heterospecific environment for extended periods (up to 21 days), the right-lateralized pattern of epidural auditory potentials first reverses transiently then returns to the typical pattern. Using acute, bilateral multi-unit electrophysiology, we confirm that this dynamic pattern occurs in NCM. Furthermore, extended exposure enhances discrimination for heterospecific stimuli. We conclude that lateralization is functionally labile and, when engaged by novel sensory experience, contributes to discrimination of novel stimuli that may be ethologically relevant. Future studies seek to determine whether, (1) the dynamicity of lateralized processes engaged by novel sensory experiences recurs with every novel challenge in the same organism; (2) the dynamic pattern extends to other cortical, thalamic or midbrain structures; and (3) the phenomenon generalizes across sensory modalities.

Statistical learning of transition patterns in the songbird auditory forebrain.

Statistical learning of transition patterns between sounds-a striking capability of the auditory system-plays an essential role in animals' survival (e.g., detect deviant sounds that signal danger). However, the neural mechanisms underlying this capability are still not fully understood. We recorded extracellular multi-unit and single-unit activity in the auditory forebrain of awake male zebra finches while presenting rare repetitions of a single sound in a long sequence of sounds (canary and zebra finch song syllables) patterned in either an alternating or random order at different inter-stimulus intervals (ISI). When preceding stimuli were regularly alternating (alternating condition), a repeated stimulus violated the preceding transition pattern and was a deviant. When preceding stimuli were in random order (control condition), a repeated stimulus did not violate any regularities and was not a deviant. At all ISIs tested (1 s, 3 s, or jittered at 0.8-1.2 s), deviant repetition enhanced neural responses in the alternating condition in a secondary auditory area (caudomedial nidopallium, NCM) but not in the primary auditory area (Field L2); in contrast, repetition suppressed responses in the control condition in both Field L2 and NCM. When stimuli were presented in the classical oddball paradigm at jittered ISI (0.8-1.2 s), neural responses in both NCM and Field L2 were stronger when a stimulus occurred as deviant with low probability than when the same stimulus occurred as standard with high probability. Together, these results demonstrate: (1) classical oddball effect exists even when ISI is jittered and the onset of a stimulus is not fully predictable; (2) neurons in NCM can learn transition patterns between sounds at multiple ISIs and detect violation of these transition patterns; (3) sensitivity to deviant sounds increases from Field L2 to NCM in the songbird auditory forebrain. Further studies using the current paradigms may help us understand the neural substrate of statistical learning and even speech comprehension.

Rapid and long-lasting improvements in neural discrimination of acoustic signals with passive familiarization.

Sensory representations in the adult brain must undergo dynamic changes to adapt to the complexity of the external world. This study investigated how passive exposure to novel sounds modifies neural representations to facilitate recognition and discrimination, using the zebra finch model organism. The neural responses in an auditory structure in the zebra finch brain, Caudal Medial Nidopallium (NCM), undergo a long-term form of adaptation with repeated stimulus presentation, providing an excellent substrate to probe the neural underpinnings of adaptive sensory representations. In Experiment 1, electrophysiological activity in NCM was recorded under passive listening conditions as novel natural vocalizations were familiarized through playback. Neural decoding of stimuli using the temporal profiles of both single-unit and multi-unit responses improved dramatically during the first few stimulus presentations. During subsequent encounters, these signals were recognized after hearing fewer initial acoustic features. Remarkably, the accuracy of neural decoding was higher when different stimuli were heard in separate blocks compared to when they were presented randomly in a shuffled sequence. NCM neurons with narrow spike waveforms generally yielded higher neural decoding accuracy than wide spike neurons, but the rate at which these accuracies improved with passive exposure was comparable between the two neuron types. Experiment 2 supported and extended these findings by showing that the rapid gains in neural decoding of novel vocalizations with passive familiarization were long-lasting, maintained for 20 hours after the initial encounter, in multi-unit responses. Taken together, these findings provide valuable insights into the mechanisms by which the nervous system dynamically modulates sensory representations to improve discrimination of novel complex signals over short and long timescales. Similar mechanisms may also be engaged during processing of human speech signals, and thus may have potential translational relevance for elucidating the neural basis of speech comprehension difficulties.

Neural Correlate of Transition Violation and Deviance Detection in the Songbird Auditory Forebrain.

Deviants are stimuli that violate one's prediction about the incoming stimuli. Studying deviance detection helps us understand how nervous system learns temporal patterns between stimuli and forms prediction about the future. Detecting deviant stimuli is also critical for animals' survival in the natural environment filled with complex sounds and patterns. Using natural songbird vocalizations as stimuli, we recorded multi-unit and single-unit activity from the zebra finch auditory forebrain while presenting rare repeated stimuli after regular alternating stimuli (alternating oddball experiment) or rare deviant among multiple different common stimuli (context oddball experiment). The alternating oddball experiment showed that neurons were sensitive to rare repetitions in regular alternations. In the absence of expectation, repetition suppresses neural responses to the 2nd stimulus in the repetition. When repetition violates expectation, neural responses to the 2nd stimulus in the repetition were stronger than expected. The context oddball experiment showed that a stimulus elicits stronger neural responses when it is presented infrequently as a deviant among multiple common stimuli. As the acoustic differences between deviant and common stimuli increase, the response enhancement also increases. These results together showed that neural encoding of a stimulus depends not only on the acoustic features of the stimulus but also on the preceding stimuli and the transition patterns between them. These results also imply that the classical oddball effect may result from a combination of repetition suppression and deviance enhancement. Classification analyses showed that the difficulties in decoding the stimulus responsible for the neural responses differed for deviants in different experimental conditions. These findings suggest that learning transition patterns and detecting deviants in natural sequences may depend on a hierarchy of neural mechanisms, which may be involved in more complex forms of auditory processing that depend on the transition patterns between stimuli, such as speech processing.

HDAC3 Inhibitor RGFP966 Modulates Neuronal Memory for Vocal Communication Signals in a Songbird Model.

Epigenetic mechanisms that modify chromatin conformation have recently been under investigation for their contributions to learning and the formation of memory. For example, the role of enzymes involved in histone acetylation are studied in the formation of long-lasting memories because memory consolidation requires gene expression events that are facilitated by an open state of chromatin. We recently proposed that epigenetic events may control the entry of specific sensory features into long-term memory by enabling transcription-mediated neuronal plasticity in sensory brain areas. Histone deacetylases, like HDAC3, may thereby regulate the specific sensory information that is captured for entry into long-term memory stores (Phan and Bieszczad, 2016). To test this hypothesis, we used an HDAC3-selective inhibitor (RGFP966) to determine whether its application after an experience with a sound stimulus with unique acoustic features could contribute to the formation of a memory that would assist in mediating its later recognition. We gave adult male zebra finches limited exposure to unique conspecific songs (20 repetitions each, well below the normal threshold to form long-term memory), followed by treatment with RGFP966 or vehicle. In different groups, we either made multi-electrode recordings in the higher auditory area NCM (caudal medial nidopallidum), or determined expression of an immediate early gene, zenk (also identified as zif268, egr-1, ngfi-a and krox24), known to participate in neuronal memory in this system. We found that birds treated with RGFP966 showed neuronal memory after only limited exposure, while birds treated with vehicle did not. Strikingly, evidence of neuronal memory in NCM induced by HDAC3-inhibition was lateralized to the left-hemisphere, consistent with our finding that RGFP966-treatment also elevated zenk expression only in the left hemisphere. The present findings show feasibility for epigenetic mechanisms to control neural plasticity underlying the formation of specific memories for conspecific communication sounds. This is the first evidence in zebra finches that epigenetic mechanisms may contribute to gene expression events for memory of acoustically-rich sensory cues.

He hears, she hears: are there sex differences in auditory processing?

Songbirds learn individually unique songs through vocal imitation and use them in courtship and territorial displays. Previous work has identified a forebrain auditory area, the caudomedial nidopallium (NCM), that appears specialized for discriminating and remembering conspecific vocalizations. In zebra finches (ZFs), only males produce learned vocalizations, but both sexes process these and other signals. This study assessed sex differences in auditory processing by recording extracellular multiunit activity at multiple sites within NCM. Juvenile female ZFs (n = 46) were reared in individual isolation and artificially tutored with song. In adulthood, songs were played back to assess auditory responses, stimulus-specific adaptation, neural bias for conspecific song, and memory for the tutor's song, as well as recently heard songs. In a subset of females (n = 36), estradiol (E2) levels were manipulated to test the contribution of E2, known to be synthesized in the brain, to auditory responses. Untreated females (n = 10) showed significant differences in response magnitude and stimulus-specific adaptation compared to males reared in the same paradigm (n = 9). In hormone-manipulated females, E2 augmentation facilitated the memory for recently heard songs in adulthood, but neither E2 augmentation (n = 15) nor E2 synthesis blockade (n = 9) affected tutor song memory or the neural bias for conspecific song. The results demonstrate subtle sex differences in processing communication signals, and show that E2 levels in female songbirds can affect the memory for songs of potential suitors, thus contributing to the process of mate selection. The results also have potential relevance to clinical interventions that manipulate E2 in human patients.

Using learned calls to study sensory-motor integration in songbirds.

Communicating songbirds produce calls as well as song and some of these are learned. One of these-the long call in zebra finches-is used by both sexes in similar behavioral contexts, but is learned in males and not in females. The male long call includes learned spectral and temporal features. In several studies, the learned long call has been used as a tool to study sensory-motor integration and vocal learning in a way that complements the use of song. Lesion studies showed that production of the male-typical call features requires an intact nucleus RA, the sexually dimorphic source of the telencephalic projection to brainstem vocal effectors. Behavioral studies that quantified zebra finch calling in response to long call playbacks showed that intact adult males have a categorical preference, absent in females, for the long calls of females over those of males. By using synthetic call stimuli, it was found that males use both spectral and temporal information to classify long call stimuli by gender, but that females use only temporal information. In juvenile males, the emergence of categorical preference occurs during the same period when RA matures anatomically (40-50 days) and the first male-typical vocalizations are produced. Adult males with RA lesions lost the categorical preference for female long calls, suggesting that RA could also play a role in long call discrimination. Preliminary analysis of recordings from neurons in NCM-a telencephalic auditory area (see Mello and colleagues, this volume)-suggests a pattern of responses to the spectral features of synthetic call stimuli that parallels the behavioral responses they elicit.

Hemispheric asymmetry in new neurons in adulthood is associated with vocal learning and auditory memory.

Many brain regions exhibit lateral differences in structure and function, and also incorporate new neurons in adulthood, thought to function in learning and in the formation of new memories. However, the contribution of new neurons to hemispheric differences in processing is unknown. The present study combines cellular, behavioral, and physiological methods to address whether 1) new neuron incorporation differs between the brain hemispheres, and 2) the degree to which hemispheric lateralization of new neurons correlates with behavioral and physiological measures of learning and memory. The songbird provides a model system for assessing the contribution of new neurons to hemispheric specialization because songbird brain areas for vocal processing are functionally lateralized and receive a continuous influx of new neurons in adulthood. In adult male zebra finches, we quantified new neurons in the caudomedial nidopallium (NCM), a forebrain area involved in discrimination and memory for the complex vocalizations of individual conspecifics. We assessed song learning and recorded neural responses to song in NCM. We found significantly more new neurons labeled in left than in right NCM; moreover, the degree of asymmetry in new neuron numbers was correlated with the quality of song learning and strength of neuronal memory for recently heard songs. In birds with experimentally impaired song quality, the hemispheric difference in new neurons was diminished. These results suggest that new neurons may contribute to an allocation of function between the hemispheres that underlies the learning and processing of complex signals.

To modulate and be modulated: estrogenic influences on auditory processing of communication signals within a socio-neuro-endocrine framework.

Gonadal hormones modulate behavioral responses to sexual stimuli, and communication signals can also modulate circulating hormone levels. In several species, these combined effects appear to underlie a two-way interaction between circulating gonadal hormones and behavioral responses to socially salient stimuli. Recent work in songbirds has shown that manipulating local estradiol levels in the auditory forebrain produces physiological changes that affect discrimination of conspecific vocalizations and can affect behavior. These studies provide new evidence that estrogens can directly alter auditory processing and indirectly alter the behavioral response to a stimulus. These studies show that: 1) Local estradiol action within an auditory area is necessary for socially relevant sounds to induce normal physiological responses in the brains of both sexes; 2) These physiological effects occur much more quickly than predicted by the classical time-frame for genomic effects; 3) Estradiol action within the auditory forebrain enables behavioral discrimination among socially relevant sounds in males; and 4) Estradiol is produced locally in the male brain during exposure to particular social interactions. The accumulating evidence suggests a socio-neuro-endocrinology framework in which estradiol is essential to auditory processing, is increased by a socially relevant stimulus, acts rapidly to shape perception of subsequent stimuli experienced during social interactions, and modulates behavioral responses to these stimuli. Brain estrogens are likely to function similarly in both songbird sexes because aromatase and estrogen receptors are present in both male and female forebrain. Estrogenic modulation of perception in songbirds and perhaps other animals could fine-tune male advertising signals and female ability to discriminate them, facilitating mate selection by modulating behaviors.

Blocking estradiol synthesis affects memory for songs in auditory forebrain of male zebra finches.

Estradiol (E2) has recently been shown to modulate sensory processing in an auditory area of the songbird forebrain, the caudomedial nidopallium (NCM). When a bird hears conspecific song, E2 increases locally in NCM, where neurons express both the aromatase enzyme that synthesizes E2 from precursors and estrogen receptors. Auditory responses in NCM show a form of neuronal memory: repeated playback of the unique learned vocalizations of conspecific individuals induces long-lasting stimulus-specific adaptation of neural responses to each vocalization. To test the role of E2 in this auditory memory, we treated adult male zebra finches (n=16) with either the aromatase inhibitor fadrozole (FAD) or saline for 8 days. We then exposed them to 'training' songs and, 6 h later, recorded multiunit auditory responses with an array of 16 microelectrodes in NCM. Adaptation rates (a measure of stimulus-specific adaptation) to playbacks of training and novel songs were computed, using established methods, to provide a measure of neuronal memory. Recordings from the FAD-treated birds showed a significantly reduced memory for the training songs compared with saline-treated controls, whereas auditory processing for novel songs did not differ between treatment groups. In addition, FAD did not change the response bias in favor of conspecific over heterospecific song stimuli. Our results show that E2 depletion affects the neuronal memory for vocalizations in songbird NCM, and suggest that E2 plays a necessary role in auditory processing and memory for communication signals.

Response properties of the auditory telencephalon in songbirds change with recent experience and season.

The caudomedial nidopallium (NCM) is a telencephalic auditory area that is selectively activated by conspecific vocalizations in zebra finches and canaries. We recently demonstrated that temporal and spectral dynamics of auditory tuning in NCM differ between these species [1]. In order to determine whether these differences reflect recent experience, we exposed separate groups of each species and sex to different housing conditions. Adult birds were housed either in an aviary with conspecifics (NORM), with heterospecifics (canary subjects in a zebra finch aviary, and vice versa: (CROSS)), or in isolation (ISO) for 9 days prior to testing. We then recorded extracellular multi-unit electrophysiological responses to simple pure tone stimuli (250-5000 Hz) in awake birds from each group and analyzed auditory tuning width using methods from our earlier studies. Relative to NORM birds, tuning was narrower in CROSS birds, and wider in ISO birds. The trend was greater in canaries, especially females. The date of recording was also included as a covariate in ANCOVAs that analyzed a larger set of the canary data, including data from birds tested outside of the breeding season, and treated housing condition and sex as independent variables. These tests show that tuning width was narrower early in the year and broader later. This effect was most pronounced in CROSS males. The degree of the short-term neural plasticity described here differs across sexes and species, and may reflect differences in NCM's anatomical and functional organization related to species differences in song characteristics, adult plasticity and/or social factors. More generally, NCM tuning is labile and may be modulated by recent experience to reflect the auditory processing required for behavioral adaptation to the current acoustic, social or seasonal context.

Inhibitory network interactions shape the auditory processing of natural communication signals in the songbird auditory forebrain.

The role of GABA in the central processing of complex auditory signals is not fully understood. We have studied the involvement of GABA A-mediated inhibition in the processing of birdsong, a learned vocal communication signal requiring intact hearing for its development and maintenance. We focused on caudomedial nidopallium (NCM), an area analogous to parts of the mammalian auditory cortex with selective responses to birdsong. We present evidence that GABA A-mediated inhibition plays a pronounced role in NCM's auditory processing of birdsong. Using immunocytochemistry, we show that approximately half of NCM's neurons are GABAergic. Whole cell patch-clamp recordings in a slice preparation demonstrate that, at rest, spontaneously active GABAergic synapses inhibit excitatory inputs onto NCM neurons via GABA A receptors. Multi-electrode electrophysiological recordings in awake birds show that local blockade of GABA A-mediated inhibition in NCM markedly affects the temporal pattern of song-evoked responses in NCM without modifications in frequency tuning. Surprisingly, this blockade increases the phasic and largely suppresses the tonic response component, reflecting dynamic relationships of inhibitory networks that could include disinhibition. Thus processing of learned natural communication sounds in songbirds, and possibly other vocal learners, may depend on complex interactions of inhibitory networks.

Species differences in auditory processing dynamics in songbird auditory telencephalon.

The caudomedial nidopallium (NCM) of songbirds is a telencephalic area involved in the auditory processing and memorization of complex vocal communication signals. We used pure tone stimuli and multiunit electrophysiological recordings in awake birds to investigate whether the basic properties of song-responding circuits in NCM differ between canaries and zebra finches, two species whose songs are markedly different in their spectral and temporal organization. We found that the responses in zebra finch NCM are characterized by broad tuning and sustained responses that may facilitate the integration of zebra finch song syllables and call notes that are of long duration and have a broad harmonic structure. In contrast, we found that the responses in canary NCM show narrower tuning and less sustained responses over the time periods analyzed. These characteristics may contribute to enhanced processing of the narrow-band whistles, rapid trills, and steep frequency modulations that are prominent features of canary song. These species differences are much less pronounced in field L2, the direct thalamorecipient region that represents a preceding station in the central avian auditory pathway. NCM responses did not differ across sexes of either species, but field L2 did show wider tuning in zebra finch females relative to males. In sum, species differences in the response properties of NCM likely reflect selectivity for the acoustic elements of each species' vocal repertoire.

Auditory topography and temporal response dynamics of canary caudal telencephalon.

To map the encoding of auditory cues in songbirds, multiunit electrophysiological responses to pure tone stimuli (250-5000 Hz) were recorded at 373 sites throughout the avian analogue of the mammalian auditory cortex in the caudal telencephalon of awake, restrained canaries. We found that a dorso-ventral tonotopic gradient from low to high frequency stimuli extends from the rostral field L2 to caudal-most caudo-medial nidopallium (NCM), similar to the frequency-dependent patterns of ZENK gene expression in canary NCM and to electrophysiological responses in other songbird species. However, response characteristics differ across the region. In field L2, responses are vigorous, phasic, and do not habituate to repeated presentations of the same stimulus. In an important subset of field L2 sites, tuning width narrows over the course of the response, which then terminates rapidly at stimulus offset. These properties are associated with inhibition at many nonpreferred frequencies and poststimulus inhibition at responsive frequencies. In contrast, NCM sites habituate to repeated sine waves, have wider tuning and lower amplitude responses, and rarely show inhibitory effects. Tuning curves in NCM are also flatter than those of field L2, and are often multipeaked. In addition, tuning width increases as the response unfolds and poststimulus excitation is often sustained in NCM. In sum, specific parts of the canary caudo-medial telencephalon differ in their response properties, suggesting differential roles in auditory processing. NCM properties, in particular, may allow for response integration across multiple spectrally varying stimulus elements, such as those that occur during birdsong.