Social experiences shape song preference learning independently of developmental exposure to song.

Communication governs the formation and maintenance of social relationships. The interpretation of communication signals depends not only on the signal's content but also on a receiver's individual experience. Experiences throughout life may interact to affect behavioural plasticity, such that a lack of developmental sensory exposure could constrain adult learning, while salient adult social experiences could remedy developmental deficits. We investigated how experiences impact the formation and direction of female auditory preferences in the zebra finch. Zebra finches form long-lasting pair bonds and females learn preferences for their mate's vocalizations. We found that after 2 weeks of cohabitation with a male, females formed pair bonds and learned to prefer their partner's song regardless of whether they were reared with ('normally reared') or without ('song-naive') developmental exposure to song. In contrast, females that heard but did not physically interact with a male did not prefer his song. In addition, previous work has found that song-naive females do not show species-typical preferences for courtship song. We found that cohabitation with a male ameliorated this difference in preference. Thus, courtship and pair bonding, but not acoustic-only interactions, strongly influence preference learning regardless of rearing experience, and may dynamically drive auditory plasticity for recognition and preference.

Machine learning and statistical classification of birdsong link vocal acoustic features with phylogeny.

Birdsong is a longstanding model system for studying evolution and biodiversity. Here, we collected and analyzed high quality song recordings from seven species in the family Estrildidae. We measured the acoustic features of syllables and then used dimensionality reduction and machine learning classifiers to identify features that accurately assigned syllables to species. Species differences were captured by the first 3 principal components, corresponding to basic frequency, power distribution, and spectrotemporal features. We then identified the measured features underlying classification accuracy. We found that fundamental frequency, mean frequency, spectral flatness, and syllable duration were the most informative features for species identification. Next, we tested whether specific acoustic features of species' songs predicted phylogenetic distance. We found significant phylogenetic signal in syllable frequency features, but not in power distribution or spectrotemporal features. Results suggest that frequency features are more constrained by species' genetics than are other features, and are the best signal features for identifying species from song recordings. The absence of phylogenetic signal in power distribution and spectrotemporal features suggests that these song features are labile, reflecting learning processes and individual recognition.

Auditory sensitivity and vocal acoustics in five species of estrildid songbirds.

Comparative studies of acoustic communication in clades with diverse signal features provide a powerful framework for testing relationships between perception and behaviour. We measured auditory sensitivity in five species of estrildid songbirds with acoustically distinct songs and tested whether differences aligned with species differences in song frequency content. Species were chosen based on phylogeny and differences in song acoustics. Behavioural audiograms were obtained using operant training and testing. Adult audiograms were compared across species and between sexes within a species. Juvenile and adult audiograms were compared in one species. The audiograms of adults reared by their own species and those reared and tutored by another species were compared in one species. Results showed that audiograms were similar across species and similar to previous reports of songbird auditory sensitivity. Species differed in the highest frequency detected and the frequency of peak sensitivity. While hearing frequency range was not correlated with song frequency bandwidth, the frequency of peak sensitivity was highly corelated with the frequency of peak energy in song. Sensitivity did not differ based on sex, age or tutoring experience. Our findings suggest that adaptations in songbird auditory sensitivity are largely constrained by shared peripheral and central encoding mechanisms, with species-specific perception appearing only at peak sensitivity.

The role of parvalbumin neurons in the evolution of skilled behaviours.

Understanding how animals display diverse and complex behaviours remains a central question in biology. A new study in PLOS Biology suggests that the emergence of clusters of parvalbumin neurons in the forebrain could reflect a convergent mechanism underlying the evolution of skilled behaviours in birds.

Mechanisms, development, and comparative perspectives on experience-dependent plasticity in social behavior.

Revealing the mechanisms underlying experience-dependent plasticity is a hallmark of behavioral neuroscience. While the study of social behavior has focused primarily on the neuroendocrine and neural control of social behaviors, the plasticity of these innate behaviors has received relatively less attention. Here, we review studies on mating-dependent changes to social behavior and neural circuitry across mammals, birds, and reptiles. We provide an overview of species similarities and differences in the effects of mating experiences on motivational and performative aspects of sexual behaviors, on sensory processing and preferences, and on the experience-dependent consolidation of sexual behavior. We also discuss recent insights into the neural mechanisms of and developmental influences on mating-dependent changes and outline promising approaches to investigate evolutionary parallels and divergences in experience-dependent plasticity.

Dopamine in the songbird auditory cortex shapes auditory preference.

Vocal communication signals can provide listeners with information about the signaler and elicit motivated responses. Auditory cortical and mesolimbic reward circuits are often considered to have distinct roles in these processes, with auditory cortical circuits responsible for detecting and discriminating sounds and mesolimbic circuits responsible for ascribing salience and modulating preference for those sounds. Here, we investigated whether dopamine within auditory cortical circuits themselves can shape the incentive salience of a vocal signal. Female zebra finches demonstrate natural preferences for vocal signals produced by males ("songs"), and we found that brief pairing of passive song playback with pharmacological dopamine manipulations in the secondary auditory cortex significantly altered song preferences. In particular, pairing passive song playback with retrodialysis of dopamine agonists into the auditory cortex enhanced preferences for less-preferred songs. Plasticity of song preferences by dopamine persisted for at least 1 week and was mediated by D1 receptors. In contrast, song preferences were not shaped by norepinephrine. In line with this, while we found that the ventral tegmental area, substantia nigra pars compacta, and locus coeruleus all project to the secondary auditory cortex, only dopamine-producing neurons in the ventral tegmental area differentially responded to preferred versus less-preferred songs. In contrast, norepinephrine neurons in the locus coeruleus increased expression of activity-dependent neural markers for both preferred and less-preferred songs. These data suggest that dopamine acting directly in sensory-processing areas can shape the incentive salience of communication signals.

Behavioral discrimination and time-series phenotyping of birdsong performance.

Variation in the acoustic structure of vocal signals is important to communicate social information. However, relatively little is known about the features that receivers extract to decipher relevant social information. Here, we took an expansive, bottom-up approach to delineate the feature space that could be important for processing social information in zebra finch song. Using operant techniques, we discovered that female zebra finches can consistently discriminate brief song phrases ("motifs") from different social contexts. We then applied machine learning algorithms to classify motifs based on thousands of time-series features and to uncover acoustic features for motif discrimination. In addition to highlighting classic acoustic features, the resulting algorithm revealed novel features for song discrimination, for example, measures of time irreversibility (i.e., the degree to which the statistical properties of the actual and time-reversed signal differ). Moreover, the algorithm accurately predicted female performance on individual motif exemplars. These data underscore and expand the promise of broad time-series phenotyping to acoustic analyses and social decision-making.

Acetylcholine in action.

The neurotransmitter acetylcholine influences how male finches perform courtship songs by acting on a region of the premotor cortex called HVC.

Auditory Selectivity for Spectral Contrast in Cortical Neurons and Behavior.

Vocal communication relies on the ability of listeners to identify, process, and respond to vocal sounds produced by others in complex environments. To accurately recognize these signals, animals' auditory systems must robustly represent acoustic features that distinguish vocal sounds from other environmental sounds. Vocalizations typically have spectral structure; power regularly fluctuates along the frequency axis, creating spectral contrast. Spectral contrast is closely related to harmonicity, which refers to spectral power peaks occurring at integer multiples of a fundamental frequency. Although both spectral contrast and harmonicity typify natural sounds, they may differ in salience for communication behavior and engage distinct neural mechanisms. Therefore, it is important to understand which of these properties of vocal sounds underlie the neural processing and perception of vocalizations.Here, we test the importance of vocalization-typical spectral features in behavioral recognition and neural processing of vocal sounds, using male zebra finches. We show that behavioral responses to natural and synthesized vocalizations rely on the presence of discrete frequency components, but not on harmonic ratios between frequencies. We identify a specific population of neurons in primary auditory cortex that are sensitive to the spectral resolution of vocal sounds. We find that behavioral and neural response selectivity is explained by sensitivity to spectral contrast rather than harmonicity. This selectivity emerges within the cortex; it is absent in the thalamorecipient region and present in the deep output region. Further, deep-region neurons that are contrast-sensitive show distinct temporal responses and selectivity for modulation density compared with unselective neurons.SIGNIFICANCE STATEMENT Auditory coding and perception are critical for vocal communication. Auditory neurons must encode acoustic features that distinguish vocalizations from other sounds in the environment and generate percepts that direct behavior. The acoustic features that drive neural and behavioral selectivity for vocal sounds are unknown, however. Here, we show that vocal response behavior scales with stimulus spectral contrast but not with harmonicity, in songbirds. We identify a distinct population of auditory cortex neurons in which response selectivity parallels behavioral selectivity. This neural response selectivity is explained by sensitivity to spectral contrast rather than to harmonicity. Our findings inform the understanding of how the auditory system encodes socially-relevant signals via detection of an acoustic feature that is ubiquitous in vocalizations.

Mechanisms of species diversity in birdsong learning.

Vocal communication is critical for social interactions across a diversity of animals. A subset of those animals, including humans and songbirds, must learn how to produce their vocal communication signals. In this issue of PLOS Biology, Wang and colleagues use genome-wide investigations of gene expression in species hybrids to uncover transcriptional networks that could influence species differences in song learning and production. We provide an overview of birdsong learning and discuss how the study by Wang and colleagues advances our understanding of mechanisms of song learning and evolution.

Emergent tuning for learned vocalizations in auditory cortex.

Vocal learners use early social experience to develop auditory skills specialized for communication. However, it is unknown where in the auditory pathway neural responses become selective for vocalizations or how the underlying encoding mechanisms change with experience. We used a vocal tutoring manipulation in two species of songbird to reveal that tuning for conspecific song arises within the primary auditory cortical circuit. Neurons in the deep region of primary auditory cortex responded more to conspecific songs than to other species' songs and more to species-typical spectrotemporal modulations, but neurons in the intermediate (thalamorecipient) region did not. Moreover, birds that learned song from another species exhibited parallel shifts in selectivity and tuning toward the tutor species' songs in the deep but not the intermediate region. Our results locate a region in the auditory processing hierarchy where an experience-dependent coding mechanism aligns auditory responses with the output of a learned vocal motor behavior.

Dopaminergic regulation of vocal-motor plasticity and performance.

Dopaminergic projections to the basal ganglia and nucleus accumbens shape the learning and plasticity of motivated behaviors across species. In songbirds, vocal learning relies on an evolutionarily specialized basal ganglia nucleus, Area X, densely innervated by dopaminergic inputs from the ventral tegmental area (VTA). While this positions songbirds as a unique model in which to tease apart the contributions of VTA dopamine neurons to vocal learning and performance, that potential has been largely underrealized. Recent breakthroughs recording from and manipulating activity of Area X projecting VTA neurons (VTAx) provide early evidence of the role of VTAx neurons to internally-guided vocal plasticity. We discuss these novel studies in the context of previous data implicating dopamine in Area X in the social modulation of song performance and how to reconcile effects on learning versus performance.

Developmental auditory exposure shapes responses of catecholaminergic neurons to socially-modulated song.

Developmental sensory experience is critical to the tuning of sensory systems and can shape perceptual abilities and their neural substrates. Neuromodulators, including catecholamines, contribute to sensory plasticity in both older and younger individuals and provide a mechanism for translating sensory experience into changes in brain and behavior. Less well known, however, is whether developmental sensory experience has lasting effects on the neuromodulatory neurons themselves. Here, we used female zebra finches to investigate the degree to which developmental auditory experience can have lasting effects on the density and sensory responsiveness of catecholamine-synthesizing neuron populations. We found that hearing courtship, but not non-courtship, song increased expression of the activity-dependent immediate early gene cFOS in dopamine neurons of the caudal ventral tegmental area (VTA) and this increase was dependent on whether females heard adult song during development. Developmental song exposure also affected the density of dopamine producing neurons in the rostral VTA. In contrast, song-evoked responses in noradrenergic neurons of the Locus Coeruleus were not affected by either developmental song exposure or the social context of the stimulus. These data highlight the lasting effects that developmental auditory experience can have in shaping both the density and sensory responsiveness of dopamine neuron populations.

fMRI Reveals a Novel Region for Evaluating Acoustic Information for Mate Choice in a Female Songbird.

Selection of sexual partners is among the most critical decisions that individuals make and is therefore strongly shaped by evolution. In social species, where communication signals can convey substantial information about the identity, state, or quality of the signaler, accurate interpretation of communication signals for mate choice is crucial. Despite the importance of social information processing, to date, relatively little is known about the neurobiological mechanisms that contribute to sexual decision making and preferences. In this study, we used a combination of whole-brain functional magnetic resonance imaging (fMRI), immediate early gene expression, and behavior tests to identify the circuits that are important for the perception and evaluation of courtship songs in a female songbird, the zebra finch (Taeniopygia guttata). Female zebra finches are sensitive to subtle differences in male song performance and strongly prefer the longer, faster, and more stereotyped courtship songs to non-courtship renditions. Using BOLD fMRI and EGR1 expression assays, we uncovered a novel region involved in auditory perceptual decision making located in a sensory integrative region of the avian central nidopallium outside the traditionally studied auditory forebrain pathways. Changes in activity in this region in response to acoustically similar but categorically divergent stimuli showed stronger parallels to behavioral responses than an auditory sensory region. These data highlight a potential role for the caudocentral nidopallium (NCC) as a novel node in the avian circuitry underlying the evaluation of acoustic signals and their use in mate choice.

Courtship song preferences in female zebra finches are shaped by developmental auditory experience.

The performance of courtship signals provides information about the behavioural state and quality of the signaller, and females can use such information for social decision-making (e.g. mate choice). However, relatively little is known about the degree to which the perception of and preference for differences in motor performance are shaped by developmental experiences. Furthermore, the neural substrates that development could act upon to influence the processing of performance features remains largely unknown. In songbirds, females use song to identify males and select mates. Moreover, female songbirds are often sensitive to variation in male song performance. Consequently, we investigated how developmental exposure to adult male song affected behavioural and neural responses to song in a small, gregarious songbird, the zebra finch. Zebra finch males modulate their song performance when courting females, and previous work has shown that females prefer the high-performance, female-directed courtship song. However, unlike females allowed to hear and interact with an adult male during development, females reared without developmental song exposure did not demonstrate behavioural preferences for high-performance courtship songs. Additionally, auditory responses to courtship and non-courtship song were altered in adult females raised without developmental song exposure. These data highlight the critical role of developmental auditory experience in shaping the perception and processing of song performance.

Singing modulates parvalbumin interneurons throughout songbird forebrain vocal control circuitry.

Across species, the performance of vocal signals can be modulated by the social environment. Zebra finches, for example, adjust their song performance when singing to females ('female-directed' or FD song) compared to when singing in isolation ('undirected' or UD song). These changes are salient, as females prefer the FD song over the UD song. Despite the importance of these performance changes, the neural mechanisms underlying this social modulation remain poorly understood. Previous work in finches has established that expression of the immediate early gene EGR1 is increased during singing and modulated by social context within the vocal control circuitry. Here, we examined whether particular neural subpopulations within those vocal control regions exhibit similar modulations of EGR1 expression. We compared EGR1 expression in neurons expressing parvalbumin (PV), a calcium buffer that modulates network plasticity and homeostasis, among males that performed FD song, males that produced UD song, or males that did not sing. We found that, overall, singing but not social context significantly affected EGR1 expression in PV neurons throughout the vocal control nuclei. We observed differences in EGR1 expression between two classes of PV interneurons in the basal ganglia nucleus Area X. Additionally, we found that singing altered the amount of PV expression in neurons in HVC and Area X and that distinct PV interneuron types in Area X exhibited different patterns of modulation by singing. These data indicate that throughout the vocal control circuitry the singing-related regulation of EGR1 expression in PV neurons may be less influenced by social context than in other neuron types and raise the possibility of cell-type specific differences in plasticity and calcium buffering.

Social context differentially modulates activity of two interneuron populations in an avian basal ganglia nucleus.

Basal ganglia circuits are critical for the modulation of motor performance across behavioral states. In zebra finches, a cortical-basal ganglia circuit dedicated to singing is necessary for males to adjust their song performance and transition between spontaneous singing, when they are alone ("undirected" song), and a performance state, when they sing to a female ("female-directed" song). However, we know little about the role of different basal ganglia cell types in this behavioral transition or the degree to which behavioral context modulates the activity of different neuron classes. To investigate whether interneurons in the songbird basal ganglia encode information about behavioral state, I recorded from two interneuron types, fast-spiking interneurons (FSI) and external pallidal (GPe) neurons, in the songbird basal ganglia nucleus area X during both female-directed and undirected singing. Both cell types exhibited higher firing rates, more frequent bursting, and greater trial-by-trial variability in firing when male zebra finches produced undirected songs compared with when they produced female-directed songs. However, the magnitude and direction of changes to the firing rate, bursting, and variability of spiking between when birds sat silently and when they sang undirected and female-directed song varied between FSI and GPe neurons. These data indicate that social modulation of activity important for eliciting changes in behavioral state is present in multiple cell types within area X and suggests that social interactions may adjust circuit dynamics during singing at multiple points within the circuit.

Sex differences and endocrine regulation of auditory-evoked, neural responses in African clawed frogs (Xenopus).

Mating depends on the accurate detection of signals that convey species identity and reproductive state. In African clawed frogs, Xenopus, this information is conveyed by vocal signals that differ in temporal patterns and spectral features between sexes and across species. We characterized spectral sensitivity using auditory-evoked potentials (AEPs), commonly known as the auditory brainstem response, in males and females of four Xenopus species. In female X. amieti, X. petersii, and X. laevis, peripheral auditory sensitivity to their species own dyad-two, species-specific dominant frequencies in the male advertisement call-is enhanced relative to males. Males were most sensitive to lower frequencies including those in the male-directed release calls. Frequency sensitivity was influenced by endocrine state; ovariectomized females had male-like auditory tuning while dihydrotestosterone-treated, ovariectomized females maintained female-like tuning. Thus, adult, female Xenopus demonstrate an endocrine-dependent sensitivity to the spectral features of conspecific male advertisement calls that could facilitate mating. Xenopus AEPs resemble those of other species in stimulus and level dependence, and in sensitivity to anesthetic (MS222). AEPs were correlated with body size and sex within some species. A frequency following response, probably encoded by the amphibian papilla, might facilitate dyad source localization via interaural time differences.

Variation in social relationships relates to song preferences and EGR1 expression in a female songbird.

Social experiences can profoundly shape social behavior and the underlying neural circuits. Across species, the formation of enduring social relationships is associated with both neural and behavioral changes. However, it remains unclear how longer-term relationships between individuals influence brain and behavior. Here, we investigated how variation in social relationships relates to variation in female preferences for and neural responses to song in a pair-bonding songbird. We assessed variation in the interactions between individuals in male-female zebra finch pairs and found that female preferences for their mate's song were correlated with the degree of affiliation and amount of socially modulated singing, but not with the frequency of aggressive interactions. Moreover, variation in measures of pair quality and preference correlated with variation in the song-induced expression of EGR1, an immediate early gene related to neural activity and plasticity, in brain regions important for auditory processing and social behavior. For example, females with weaker preferences for their mate's song had greater EGR1 expression in the nucleus Taeniae, the avian homologue of the mammalian medial amygdala, in response to playback of their mate's courtship song. Our data indicate that the quality of social interactions within pairs relates to variation in song preferences and neural responses to ethologically relevant stimuli and lend insight into neural circuits sensitive to social information. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 76: 1029-1040, 2016.