Tutor Song Research Articles

A Synaptic Basis for Auditory–Vocal Integration in the Songbird

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

Regulation of forebrain perineuronal nets during a critical period for vocal learning

Event Abstract Back to Event Regulation of forebrain perineuronal nets during a critical period for vocal learning Teresa A. Nick1*, T. S. Balmer1, V. M. Carels1 and J. L. Frisch1 1 University of Minnesota Medical School, United States Extracellular matrix structures known as perineuronal nets (PNNs) appear to have roles in the closure of sensory critical periods and may define the critical period for ocular dominance plasticity in the primary visual cortex (Hensch, 2005; Pizzorusso et al., 2002). We hypothesized that similar mechanisms may also underlie sensorimotor critical periods, such as that for speech. For our model, we chose the zebra finch songbird (Taeniopygia guttata). As in humans, vocal sensorimotor learning in the zebra finch occurs during a well-defined and restricted developmental period. The finch learns a song from an adult tutor and then gradually matches the song he produces to the memorized tutor song. Song learning and production depend on a set of anatomically distinct forebrain song nuclei, which allows precise investigation of brain areas with defined behavioral roles. PNNs in song nuclei were examined at the beginning, middle, and end of the sensorimotor learning phase using antibodies to chondroitin sulfate, which composes the PNNs. We found that PNNs are strongly expressed in the adult song system (Fig. 1).PNNs in mammals are preferentially expressed around typically fast-spiking neurons that express parvalbumin, a calcium-binding protein. Finch brain sections were thus also stained for parvalbumin. All neurons in each nucleus were identified using a fluorescent Nissl stain. Consistent with a role for PNNs in vocal development, both the percent of neurons with PNNs and the percent of parvalbumin-positive neurons with PNNs increased to adult levels during the sensorimotor period of vocal development in all song nuclei examined. Stereological quantification confirmed these findings. Several perturbations alter vocal learning, such as overstimulation with tutor song auditory playback, isolation from a song tutor, and chronic exposure to testosterone. Preliminary data suggest that experimentally-induced shifts in the vocal critical period also shift PNN expression pattern. These data suggest that perineuronal nets may regulate plasticity in forebrain networks that underlie sensorimotor learning. Conference: 10th International Conference on Cognitive Neuroscience, Bodrum, Türkiye, 1 Sep - 5 Sep, 2008. Presentation Type: Poster Presentation Topic: Neuronal Plasticity Citation: Nick TA, Balmer TS, Carels VM and Frisch JL (2008). Regulation of forebrain perineuronal nets during a critical period for vocal learning. Conference Abstract: 10th International Conference on Cognitive Neuroscience. doi: 10.3389/conf.neuro.09.2009.01.311 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 10 Dec 2008; Published Online: 10 Dec 2008. * Correspondence: Teresa A Nick, University of Minnesota Medical School, Minneapolis, United States, nickx002@umn.edu Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Teresa A Nick T. S Balmer V. M Carels J. L Frisch Google Teresa A Nick T. S Balmer V. M Carels J. L Frisch Google Scholar Teresa A Nick T. S Balmer V. M Carels J. L Frisch PubMed Teresa A Nick T. S Balmer V. M Carels J. L Frisch Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Incomplete and Inaccurate Vocal Imitation after Knockdown of FoxP2 in Songbird Basal Ganglia Nucleus Area X

The gene encoding the forkhead box transcription factor, FOXP2, is essential for developing the full articulatory power of human language. Mutations of FOXP2 cause developmental verbal dyspraxia (DVD), a speech and language disorder that compromises the fluent production of words and the correct use and comprehension of grammar. FOXP2 patients have structural and functional abnormalities in the striatum of the basal ganglia, which also express high levels of FOXP2. Since human speech and learned vocalizations in songbirds bear behavioral and neural parallels, songbirds provide a genuine model for investigating the basic principles of speech and its pathologies. In zebra finch Area X, a basal ganglia structure necessary for song learning, FoxP2 expression increases during the time when song learning occurs. Here, we used lentivirus-mediated RNA interference (RNAi) to reduce FoxP2 levels in Area X during song development. Knockdown of FoxP2 resulted in an incomplete and inaccurate imitation of tutor song. Inaccurate vocal imitation was already evident early during song ontogeny and persisted into adulthood. The acoustic structure and the duration of adult song syllables were abnormally variable, similar to word production in children with DVD. Our findings provide the first example of a functional gene analysis in songbirds and suggest that normal auditory-guided vocal motor learning requires FoxP2.

Model of Birdsong Learning Based on Gradient Estimation by Dynamic Perturbation of Neural Conductances

We propose a model of songbird learning that focuses on avian brain areas HVC and RA, involved in song production, and area LMAN, important for generating song variability. Plasticity at HVC --> RA synapses is driven by hypothetical "rules" depending on three signals: activation of HVC --> RA synapses, activation of LMAN --> RA synapses, and reinforcement from an internal critic that compares the bird's own song with a memorized template of an adult tutor's song. Fluctuating glutamatergic input to RA from LMAN generates behavioral variability for trial-and-error learning. The plasticity rules perform gradient-based reinforcement learning in a spiking neural network model of song production. Although the reinforcement signal is delayed, temporally imprecise, and binarized, the model learns in a reasonable amount of time in numerical simulations. Varying the number of neurons in HVC and RA has little effect on learning time. The model makes specific predictions for the induction of bidirectional long-term plasticity at HVC --> RA synapses.

Song Selectivity in the Pallial-Basal Ganglia Song Circuit of Zebra Finches Raised Without Tutor Song Exposure

Acoustic experience critically influences auditory cortical development as well as emergence of highly selective auditory neurons in the songbird sensorimotor circuit. In adult zebra finches, these "song-selective" neurons respond better to the bird's own song (BOS) than to songs of other conspecifics. Birds learn their songs by memorizing a tutor's song and then matching auditory feedback of their voice to the tutor song memory. Song-selective neurons in the pallial-basal ganglia circuit called the anterior forebrain pathway (AFP) reflect the development of BOS. However, during learning, they also respond strongly to tutor song and are compromised in their adult selectivity when birds are prevented from matching BOS to tutor, suggesting that selectivity depends on tutor song learning as well as sensorimotor matching of BOS feedback to the tutor song memory. We examined the contribution of sensory learning of tutor song to song selectivity by recording from AFP neurons in birds reared without exposure to adult conspecifics. We found that AFP neurons in these "isolate" birds had highly tuned responses to isolate BOS. The selectivity was as high, and in the striato-pallidal nucleus Area X, even higher than that in normal birds, due to abnormally weak responsiveness to conspecific song. These results demonstrate that sensory learning of tutor song is not necessary for BOS tuning of AFP neurons. Because isolate birds develop their song via sensorimotor learning, our data further illustrate the importance of individual sensorimotor learning for song selectivity and provide insight into possible functions of song-selective neurons.

Functional MRI of the zebra finch brain during song stimulation suggests a lateralized response topography.

Electrophysiological and activity-dependent gene expression studies of birdsong have contributed to the understanding of the neural representation of natural sounds. However, we have limited knowledge about the overall spatial topography of song representation in the avian brain. Here, we adapt the noninvasive functional MRI method in mildly sedated zebra finches (Taeniopygia guttata) to localize and characterize song driven brain activation. Based on the blood oxygenation level-dependent signal, we observed a differential topographic responsiveness to playback of bird's own song, tutor song, conspecific song, and a pure tone as a nonsong stimulus. The bird's own song caused a stronger response than the tutor song or tone in higher auditory areas. This effect was more pronounced in the medial parts of the forebrain. We found left-right hemispheric asymmetry in sensory responses to songs, with significant discrimination between stimuli observed only in the right hemisphere. This finding suggests that perceptual responses might be lateralized in zebra finches. In addition to establishing the feasibility of functional MRI in sedated songbirds, our results demonstrate spatial coding of song in the zebra finch forebrain, based on developmental familiarity and experience.

Adult and peer influences on starling song development

Adult influence on song development was rarely investigated in complex social settings. We investigated the influence of the young/adult ratio on song learning in European starlings. Starling songs are composed of two categories: whistles and warbling. Warbling includes three parts: complex variable motifs, click motifs, and high frequency motifs. Song learning by young birds raised in group (G birds), in group with adults (GT birds) or in pair with one adult (IT birds) were compared. Song tutor choice, song sharing, and song quality varied with social context. The mere presence of adults in a group was not sufficient for young to develop all adult song features. Only IT birds chose adults as tutors. Social contact with only one adult enhanced acquisition of whistles, click motifs, individuality, and reduced the number of high frequency motifs (adult song features). This study suggests that the adult influence is proportional to young/adult ratio.

Birdsong Memory: A Neural Dissociation between Song Recognition and Production

Songbirds learn their song from an adult conspecific tutor when they are young, much like the acquisition of speech in human infants. When an adult zebra finch is re-exposed to its tutor's song, there is increased neuronal activation in the caudomedial nidopallium (NCM), the songbird equivalent of the auditory association cortex. This neuronal activation is related to the fidelity of song imitation, suggesting that the NCM may contain the neural representation of song memory. We found that bilateral neurotoxic lesions to the NCM of adult male zebra finches impaired tutor-song recognition but did not affect the males' song production or their ability to discriminate calls. These findings demonstrate that the NCM performs an essential role in the representation of tutor-song memory. In addition, our results show that tutor-song memory and a motor program for the bird's own song have separate neural representations in the songbird brain. Thus, in both humans and songbirds, the cognitive systems of vocal production and auditory recognition memory are subserved by distinct brain regions.

HVC Neural Sleep Activity Increases With Development and Parallels Nightly Changes in Song Behavior

Sleep abnormalities are coexpressed with human communication disorders. Recent data from the birdsong system, the best model for human speech, indicate that sleep has a critical role in vocal learning. To understand the neural mechanisms that underlie behavioral changes during sleep, we recorded sleep activity in the song control area HVC longitudinally during song development in zebra finches. We focused on the sensorimotor phase of song learning, when the finch shapes his song behavior toward a learned tutor song model. Direct comparison of sleep activity in adults and juveniles revealed that the juvenile HVC has a lower spike rate and longer silent periods than the adult. Within individual finches, sleep silent periods decreased and spike rate increased with age. We next systematically compared neural sleep activity and song behavior. We now report that spike rate during sleep was significantly correlated with overnight changes in song behavior. Collectively, these data indicate that sleep activity in the vocal control area HVC increases with age and may affect song behavior.

Development of Selectivity for Natural Sounds in the Songbird Auditory Forebrain

In adult songbirds, auditory neurons in the primary auditory forebrain region of field L and a secondary auditory forebrain region of caudal mesopallium (CM) are highly responsive to natural sounds, such as conspecific song. Because these nuclei are involved in sensory representations of songs, we investigated how their function changes during development. We recorded neural responses to conspecific and tutor song and acoustically matched synthetic sounds in field L and lateral CM (CLM) of urethane-anesthetized juvenile male zebra finches of approximately 35 days of age. At this age, juvenile songbirds are memorizing the songs of their adult tutors but do not yet sing mature song. They are also starting to recognize songs of individual conspecifics. Compared with adult auditory forebrain neurons, juvenile neurons in field L were on average less responsive to auditory stimuli and exhibited less selectivity for natural sounds compared with the synthetic sounds. This developmental effect was more pronounced in the secondary subregions of L1 and L3 than in the primary thalamo-recipient subregion L2 of field L. CLM showed adultlike selectivity for natural sounds. Also, we did not find any evidence of memory for the tutor song in either field L or CLM. We note that the neural development of selective responses to conspecific song in the secondary subregions of field L is correlated with the emergence of individual song preference around 35 days of age. Therefore we suggest that the emergence of natural sound selectivity in field L could be important for the behavioral development of song recognition.

Syllable chunking in Bengalese finches in a multi-tutor environment

Experiments in birdsong learning have traditionally involved one tutor, usually the father, and the proportion and accuracy of note copying are discussed as indexes of song learning. But in the wild, there are several conspecific males around song-learning pupils. To test song learning under naturalistic situation, 11 male and ten female Bengalese finches were kept in a large flight cage containing ten pot-shaped nests. Seventy-eight chicks were born in this aviary over an 18-month period. We recorded the full songs of male birds after they had matured, and compared the songs between offspring and the original adult males. Most of them learned their songs from one or two tutors and some learned from three or four tutors. When a bird learned from multiple tutors, each of these tutor songs was copied in chunks and within a chunk the order of the song elements was the same as that observed in the tutor male. Furthermore, Bengalese finches change the order of chunks as they sing, and it was possible to express the rule using a finite-state syntax. Therefore, Bengalese finches have an innate universal grammar allowing them to construct a finite-state syntax from multiple sources. [Work supported by JSPS and JST.]

Effects of social rearing conditions on song structure and repertoire size: experimental evidence from the field

Through a cross-fostering experiment, we studied song learning of blue tit, Cyanistes caeruleus, and great tit, Parus major, males reared by heterospecific parents. This was done in the wild, and so potential song tutors, territorial neighbours, potential mates and other social factors were all natural and not affected by the treatment. Normally, the song repertoires of the two species are completely discrete. However, the cross-fostered great tit males altered their song in several aspects, including repertoire size and composition, and temporal and frequency parameters, thus becoming intermediate between the normal songs of the two species. For the cross-fostered blue tit song, only repertoire composition was affected. However, an analysis of repertoire size in both species proved this to be larger in cross-fostered than in control males. This increase in repertoire size shows that repertoire size is influenced by social conditions in these species and is not strictly constrained by memory capacity. Furthermore, several of the cross-fostered great tit males included trilled songs in their repertoires. In these two species, trilled songs have been regarded as unique to the blue tit. In conclusion, our results show the importance of early social experiences for song learning in these wild tits.

The Onset of Song Learning and Song Tutor Selection in Fledgling Zebra Finches

AbstractWe investigated song development in the pre‐independent zebra finch (aged 15–35 d), a period when neural pathways for song learning and production are forming and social influences outside the family are limited. Expt 1 investigated the onset and the minimum duration of tutoring needed for song learning in fledglings. We found most begin to learn song from 25 d of age and need about 10 d contact with the father tutor to make accurate copies. This onset corresponds with major developments in the formation of the neural circuitry implicated in song acquisition. Subsong also begins on day 25 suggesting that the sensory and motor phases of song learning fully overlap in the zebra finch. Our findings support the hypothesis that the song circuitry is fully functional by 35 d of age and the sensitive phase for zebra finches extends therefore from about days 25–65. However, only the first 10 d of this period are necessary to learn a tutor's song with fair accuracy. Expt 2 investigated the role of the paternal bond, spatial proximity and mating status in a fledgling's choice of song tutor where the father was the sole parent. Young chose the father over single unrelated males (expt 2a) or unrelated males in company with their female partners (expt 2b). Given the close spatial proximity of both potential tutors to the fledglings it is probably the filial bond, established via paternal care that is the cause of this preference. Zebra finches sing the same song phrase in two contrasting contexts: female‐directed song during pre‐copulatory courtship and undirected song where no female or display is involved. In expt 3 we tested the song learning preference of pre‐independent young for two categories of non‐paternal tutors: those singing predominantly female‐directed song and those singing exclusively undirected song. There was a small, but significant, preference for fledgling zebra finches to copy songs from males that sang female‐directed song. This preference is consistent with the hypothesis that young males not only learn the acoustic features of their tutor's song but also the visual and dynamic movements that constitute the courtship display.

Localized brain activation specific to auditory memory in a female songbird

Song acquisition in songbird males is a prominent model system for the study of the brain mechanisms of memory. Male zebra finches (Taeniopygia guttata) learn their songs from an adult conspecific tutor early in life. Previous work has shown that exposure of males to their tutor song leads to increased expression of immediate early genes (IEGs) in the caudomedial nidopallium (NCM) and in the caudomedial mesopallium (CMM). In addition, IEG expression in the NCM correlates significantly with the strength of song learning. Interpretation of these findings is complicated, as males both learn the characteristics of tutor song and learn to produce a similar own song. Female zebra finches do not sing, but nevertheless they learn the characteristics of a song to which they were exposed when young, and form a preference for it. Here, adult zebra finch females reared with their fathers showed a significant preference for their father's song. Females that were later reexposed to their father's song showed significantly greater expression of Zenk, the protein product of the IEG ZENK, than controls that were exposed to a novel song, in the CMM, but not in the NCM or hippocampus. These results suggest that in female zebra finches the CMM may be (part of) the neural substrate for the representation of the memory of their father's song.

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.

Auditory Plasticity: Vocal Output Shapes Auditory Cortex

Studies in humans and songbirds have revealed a close link between vocal output and hearing. Now experiments in marmosets have shown that self-generated vocalizations can modulate the activity of neurons in the auditory cortex and even remodel their response properties.

Freedom and Rules: The Acquisition and Reprogramming of a Bird's Learned Song

Canary song is hierarchically structured: Short stereotyped syllables are repeated to form phrases, which in turn are arranged to form songs. This structure occurs even in the songs of young isolates, which suggests that innate rules govern canary song development. However, juveniles that had never heard normal song imitated abnormal synthetic songs with great accuracy, even when the tutor songs lacked phrasing. As the birds matured, imitated songs were reprogrammed to form typical canary phrasing. Thus, imitation and innate song constraints are separate processes that can be segregated in time: freedom in youth, rules in adulthood.

Vocal Experimentation in the Juvenile Songbird Requires a Basal Ganglia Circuit

Songbirds learn their songs by trial-and-error experimentation, producing highly variable vocal output as juveniles. By comparing their own sounds to the song of a tutor, young songbirds gradually converge to a stable song that can be a remarkably good copy of the tutor song. Here we show that vocal variability in the learning songbird is induced by a basal-ganglia-related circuit, the output of which projects to the motor pathway via the lateral magnocellular nucleus of the nidopallium (LMAN). We found that pharmacological inactivation of LMAN dramatically reduced acoustic and sequence variability in the songs of juvenile zebra finches, doing so in a rapid and reversible manner. In addition, recordings from LMAN neurons projecting to the motor pathway revealed highly variable spiking activity across song renditions, showing that LMAN may act as a source of variability. Lastly, pharmacological blockade of synaptic inputs from LMAN to its target premotor area also reduced song variability. Our results establish that, in the juvenile songbird, the exploratory motor behavior required to learn a complex motor sequence is dependent on a dedicated neural circuit homologous to cortico-basal ganglia circuits in mammals.

Song tutoring triggers CaMKII phosphorylation within a specialized portion of the avian basal ganglia

In several songbird species, a specialized anterior forebrain pathway (AFP) that includes part of the avian basal ganglia has been implicated specifically in song learning. To further elucidate cellular mechanisms and circuitry involved in vocal learning, we used quantitative immunoblot analysis to determine if early song tutoring promotes within the AFP phosphorylation of calcium/calmodulin-dependent kinase II (CaMKII), a multifunctional kinase whose phosphorylation at threonine 286 is critical for many forms of neural plasticity and behavioral learning. We report that in young male zebra finches likely to have begun the process of song acquisition, brief tutoring by a familiar conspecific adult promotes a dramatic increase in levels of phosphorylated CaMKII (pCaMKII) in Area X, the striatal/pallidal component of the AFP. In contrast, pCaMKII levels in this region were not elevated if 1) the tutor did not sing, 2) the tutor sang but was visually isolated from the pupil, or 3) the tutor was an unfamiliar adult. In young males that had not previously heard any conspecific song, first exposure to a song tutor produced a more modest, but significant rise in pCaMKII levels. Young females (who do not develop song behavior) did not exhibit any effect of tutoring on pCaMKII levels in that portion of the basal ganglia that corresponds to Area X in males. These data are consistent with the hypothesis that Area X participates in encoding and/or attaching reward value to a representation of tutor song that is accessed later to guide motor learning.

Sequential Learning From Multiple Tutors and Serial Retuning of Auditory Neurons in a Brain Area Important to Birdsong Learning

Songbirds hear many vocal models during a juvenile sensitive period, transiently imitating some while retaining imitations of others in their repertoires. Despite subsequent conflicting experiences, early experience can exert lasting effects on neural structure and function, raising the possibility that transiently expressed vocalizations or their relevant models are stored in the adult songbird's brain. One site where learned song representations could be stored is the lateral magnocellular nucleus of the anterior nidopallium (LMAN), which in the adult songbird contains neurons responsive to playback of the bird's own song (BOS) and the tutor song (TS). To test whether LMAN neurons develop and retain responses to transiently learned songs, we exposed zebra finch hatchlings [posthatch day 0 (PHD0)] to a first TS (TS1) for about 30 days, isolated them for about 30 days, then exposed them to a second TS (TS2) for 30 days starting at PHD 60. Behavioral analysis showed that PHD 60 juveniles had started to copy TS1, although this copying was transient, because the adult BOS resembled TS2 and not TS1. We found that LMAN auditory responses paralleled these behavioral changes: LMAN neurons at PHD 60 responded strongly and selectively to both the juvenile BOS and TS1, whereas LMAN neurons in adults responded to the adult BOS and TS2, but not to the transiently learned song or its model. Therefore LMAN auditory responses can be lost or overwritten as the juvenile copies a new song, suggesting that the adult LMAN does not store information about transiently learned songs or their models.