Song Stimulation Research Articles

Aromatase and nonaromatase neurons in the zebra finch secondary auditory forebrain are indistinct in their song-driven gene induction and intrinsic electrophysiological properties.

Estrogens support major brain functions including cognition, reproduction, neuroprotection and sensory processing. Neuroestrogens are synthesized within some brain areas by the enzyme aromatase and can rapidly modulate local circuit functions, yet the cellular physiology and sensory-response profiles of aromatase neurons are essentially unknown. In songbirds, social and acoustic stimuli drive neuroestrogen elevations in the auditory forebrain caudomedial nidopallium (NCM). In both males and females, neuroestrogens rapidly enhance NCM auditory processing and auditory learning. Estrogen-producing neurons in NCM may therefore exhibit distinguishing profiles for sensory-activation and intrinsic electrophysiology. Here, we explored these questions using both immunocyctochemistry and electrophysiological recordings. Immunoreactivity for aromatase and the immediate early gene EGR1, a marker of activity and plasticity, were quantified in NCM of song-exposed animals versus silence-exposed controls. Using whole-cell patch clamp recordings from NCM slices, we also documented the intrinsic excitability profiles of aromatase-positive and aromatase-negative neurons. We observed that a subset of aromatase neurons were significantly activated during song playback, in both males and females, and in both hemispheres. A comparable population of non-aromatase-expressing neurons were also similarly driven by song stimulation. Membrane properties (i.e., resting membrane potential, rheobase, input resistance and multiple action potential parameters) were similarly indistinguishable between NCM aromatase and non-aromatase neurons. Together, these findings demonstrate that aromatase and non-aromatase neurons in NCM are indistinct in terms of their intrinsic electrophysiology and responses to song. Nevertheless, such similarities in response properties may belie more subtle differences in underlying conductances and/or computational roles that may be crucial to their function.

STIMULASI LAGU DALAM PENGEMBANGAN SENI ANAK USIA DINI

This research is motivated by the importance of providing song stimulation in the development of early childhood arts, while in reality the provision of song stimulation by teachers in schools still limited due to the limited production of children's thematic songs. So that it takes the teacher's ability to create simple songs according to the thematic needs of learning. This study aims to provide the elements of children's songs, namely melody, harmony, rhythm, lyrics, and tempo. Type of research used is literature study. Data were collected by reviewing and reviewing several relevant reading sources, such as scientific journals and related reference books. Furthermore, the elements that must be considered in early childhood songs are analyzed. From the research, the results show that the lyrical elements of children's songs must be positive and simple, the range and jump of the melody is right, the use of basic harmony starts and ends with level I chords, the use of a 1/4 or 1/8 rhythm, and the tempo according to the impression of the song to be shown. Thus the teacher can continue to provide song stimulation in the development of early childhood arts.

Functional genomic analysis and neuroanatomical localization of miR-2954, a song-responsive sex-linked microRNA in the zebra finch.

Natural experience can cause complex changes in gene expression in brain centers for cognition and perception, but the mechanisms that link perceptual experience and neurogenomic regulation are not understood. MicroRNAs (miRNAs or miRs) have the potential to regulate large gene expression networks, and a previous study showed that a natural perceptual stimulus (hearing the sound of birdsong in zebra finches) triggers rapid changes in expression of several miRs in the auditory forebrain. Here we evaluate the functional potential of one of these, miR-2954, which has been found so far only in birds and is encoded on the Z sex chromosome. Using fluorescence in situ hybridization and immunohistochemistry, we show that miR-2954 is present in subsets of cells in the sexually dimorphic brain regions involved in song production and perception, with notable enrichment in cell nuclei. We then probe its regulatory function by inhibiting its expression in a zebra finch cell line (G266) and measuring effects on endogenous gene expression using Illumina RNA sequencing (RNA-seq). Approximately 1000 different mRNAs change in expression by 1.5-fold or more (adjusted p < 0.01), with increases in some but not all of the targets that had been predicted by Targetscan. The population of RNAs that increase after miR-2954 inhibition is notably enriched for ones involved in the MAP Kinase (MAPK) pathway, whereas the decreasing population is dominated by genes involved in ribosomes and mitochondrial function. Since song stimulation itself triggers a decrease in miR-2954 expression followed by a delayed decrease in genes encoding ribosomal and mitochondrial functions, we suggest that miR-2954 may mediate some of the neurogenomic effects of song habituation.

Novel song-stimulated dendritic spine formation and Arc/Arg3.1 expression in zebra finch auditory telencephalon are disrupted by cannabinoid agonism

Cannabinoids are well-established to alter processes of sensory perception; however neurophysiological mechanisms responsible remain unclear. Arc, an immediate-early gene (IEG) product involved in dendritic spine dynamics and necessary for plasticity changes such as long-term potentiation, is rapidly induced within zebra finch caudal medial nidopallium (NCM) following novel song exposure, a response that habituates after repeated stimuli. Arc appears unique in its rapid postsynaptic dendritic expression following excitatory input. Previously, we found that vocal development-altering cannabinoid treatments are associated with elevated dendritic spine densities in motor-(HVC) and learning-related (Area X) song regions of zebra finch telencephalon. Given Arc's dendritic morphological role, we hypothesized that cannabinoid-altered spine densities may involve Arc-related signaling. To test this, we examined the ability of the cannabinoid agonist WIN55212-2 (WIN) to (1) acutely disrupt song-induced Arc expression, (2) interfere with habituation to auditory stimuli, and (3) alter dendritic spine densities in auditory regions. We found that WIN (3mg/kg) acutely reduced Arc expression within both NCM and Field L2 in an antagonist-reversible manner. WIN did not alter Arc expression in thalamic auditory relay nucleus ovoidalis (Ov), suggesting that cannabinoid signaling selectively alters responses to auditory stimulation. Novel song stimulation rapidly increased dendritic spine densities within auditory telencephalon, an effect blocked by WIN pretreatments. Taken together, cannabinoid inhibition of both Arc induction and its habituation to repeated stimuli, combined with prevention of rapid increases in dendritic spine densities, implicates cannabinoid signaling in modulation of physiological processes important to auditory responsiveness and memory.

Neural systems for speech and song in autism

Despite language disabilities in autism, music abilities are frequently preserved. Paradoxically, brain regions associated with these functions typically overlap, enabling investigation of neural organization supporting speech and song in autism. Neural systems sensitive to speech and song were compared in low-functioning autistic and age-matched control children using passive auditory stimulation during functional magnetic resonance and diffusion tensor imaging. Activation in left inferior frontal gyrus was reduced in autistic children relative to controls during speech stimulation, but was greater than controls during song stimulation. Functional connectivity for song relative to speech was also increased between left inferior frontal gyrus and superior temporal gyrus in autism, and large-scale connectivity showed increased frontal–posterior connections. Although fractional anisotropy of the left arcuate fasciculus was decreased in autistic children relative to controls, structural terminations of the arcuate fasciculus in inferior frontal gyrus were indistinguishable between autistic and control groups. Fractional anisotropy correlated with activity in left inferior frontal gyrus for both speech and song conditions. Together, these findings indicate that in autism, functional systems that process speech and song were more effectively engaged for song than for speech and projections of structural pathways associated with these functions were not distinguishable from controls.

The genomic response to courtship song stimulation in female Drosophila melanogaster.

Courtship behaviour involves a complex exchange of signals and responses. These are usually studied at the phenotypic level, and genetic or transcriptional responses to courtship are still poorly understood. Here, we examine the gene-expression changes in Drosophila melanogaster females in response to one of the key male courtship signals in mate recognition, song produced by male wing vibration. Using long oligonucleotide microarrays, we identified several genes that responded differentially to the presence or absence of acoustic courtship stimulus. These changes were modest in both the number of genes involved and fold-changes, but notably dominated by antennal signalling genes involved in olfaction as well as neuropeptides and immune response genes. Second, we compared the expression patterns of females stimulated with synthetic song typical of either conspecific or heterospecific (Drosophila simulans) males. In this case, antennal olfactory signalling and innate immunity genes were also enriched among the differentially expressed genes. We confirmed and investigated the time course of expression differences of two identified immunity genes using real-time quantitative PCR. Our results provide novel insight into specific molecular changes in females in response to courtship song stimulation. These may be involved in both signal perception and interpretation and some may anticipate molecular interactions that occur between the sexes after mating.

Partial dissociation of molecular and behavioral measures of song habituation in adult zebra finches

Initial playback of recorded birdsong triggers a number of responses in zebra finches, including overt listening behavior and ERK pathway-dependent activation of zenk gene transcription in the auditory lobule of the forebrain. Repetition of one song stimulus leads to persistent habituation of these responses, as measured by subsequent presentations 1 day later. In this study, we examined the causal relationships between behavioral and molecular (ERK/zenk) habituation. In a within-subject comparison, we found a strong correlation with the level of prior training for both responses (duration of behavioral listening and magnitude of zenk expression), but little correlation between these responses for birds within the same treatment group. We then tested the hypothesis that ERK/zenk activation during training is necessary for the development of habituation measured 1 day later. Cannula-directed infusion of a pharmacological inhibitor of ERK activation (U0126) immediately before training blocked the development of habituation of the zenk gene response. However, measurement of the effect on behavioral habituation was confounded because birds that were infused with a non-active drug analogue (U0124) showed a decreased response 1 day later, even to novel songs. We conclude that the behavioral response to song stimulation is strongly influenced by factors other than song familiarity, whereas the zenk response in the forebrain may be a more accurate indicator of actual experience hearing a particular song.

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.

Dynamic Role of Postsynaptic Caspase-3 and BIRC4 in Zebra Finch Song-Response Habituation

Activation of the protease caspase-3 is commonly thought to cause apoptotic cell death. Here, we show that caspase-3 activity is regulated at postsynaptic sites in brain following stimuli associated with memory (neural activation and subsequent response habituation) instead of cell death. In the zebra finch auditory forebrain, the concentration of caspase-3 active sites increases briefly within minutes after exposure to tape-recorded birdsong. With confocal and immunoelectron microscopy, we localize the activated enzyme to dendritic spines. The activated caspase-3 protein is present even in unstimulated brain but bound to an endogenous inhibitor, BIRC4 (xIAP), suggesting a mechanism for rapid release and sequestering at specific synaptic sites. Caspase-3 activity is necessary to consolidate a persistent physiological trace of the song stimulus, as demonstrated using pharmacological interference and the zenk gene habituation assay. Thus, the brain appears to have adapted a core component of cell death machinery to serve a unique role in learning and memory.

Calbindin‐positive neurons reveal a sexual dimorphism within the songbird analogue of the mammalian auditory cortex

The oscine song system, a set of interconnected brain nuclei involved in song production and learning, is one of the first and clearest examples of brain sexual dimorphism in a vertebrate, being typically well-developed in males, but not females. Here we present evidence for a sexual dimorphism in the caudomedial nidopallidum (NCM), an auditory area outside of the song system. NCM is thought to correspond to a portion of the auditory cortex of mammals and is involved in the perceptual processing of birdsong. We show that cells immunolabeled for the calcium-binding protein calbindin are primarily localized to caudal NCM and are almost twice as numerous in males as in females. We demonstrate that calbindin-positive cells constitute a subset of GABAergic cells in NCM, and show that the sex dimorphism in this cell population does not result from local gender differences in the overall density of neuronal or GABAergic cells. In addition, we demonstrate that calbindin-positive cells lack song-induced expression of the activity-dependent gene ZENK, and that song stimulation does not change the density or distribution of these cells in NCM. Finally, we show that the distribution of calbindin-positive cells in NCM is strikingly similar to the mRNA expression for the estrogen-generating enzyme aromatase. Together these results suggest that NCM is likely composed of neurochemically-distinct domains and presents a marked sex dimorphism in a specific subset of GABAergic neurons, which may confer sex-specific sensory processing capabilities to this auditory area. Our results also suggest that local sex steroid hormones may play a local role in auditory processing in the songbird telencephalon.

Rhythmic Activity in a Forebrain Vocal Control NucleusIn Vitro

The learned vocalizations of songbirds constitute a rhythmic behavior that is thought to be governed by a central pattern generator and that is accompanied by highly patterned neural activity. Phasic premotor activity is observed during singing in HVC [used as a proper name following the nomenclature of Reiner et al. (2004)], a telencephalic song system nucleus that is essential for song production. Moreover, HVC neurons display phasic patterns of auditory activity in response to song stimulation. To address the cellular basis of pattern generation in HVC, we investigated its rhythm-generating abilities. We report here the induction of sustained, rhythmic activity patterns in HVC when isolated in vitro. Brief, high-frequency stimulation evoked repetitive postsynaptic potentials (PSPs) and local field potentials (LFPs) from HVC neurons recorded in a brain slice preparation made from adult male zebra finches. These rhythmic events were sustained for seconds in the absence of ongoing, phasic stimulation, and they had temporal properties similar to those of syllable occurrence within zebra finch song. Paired recordings revealed synchrony between repetitive PSP and LFP occurrence, indicating that a population of cells participates in this patterned activity. The PSPs resulted from multiple, coordinated, fast-glutamatergic, synaptic inputs. Moreover, their occurrence and timing relied on inhibitory synaptic transmission. Thus, HVC itself has rhythmic abilities that could influence the timing of neural activity over relatively long time windows. These rhythmic properties may contribute to song production or perception in vivo.

Activation and habituation of extracellular signal-regulated kinase phosphorylation in zebra finch auditory forebrain during song presentation.

The sound of tape-recorded birdsong triggers a set of behavioral and physiological responses in zebra finches, including transcriptional activation of the zenk gene in the auditory forebrain. Song repetition leads to the stimulus-specific habituation of these responses. To gain insight into the mechanisms that couple auditory experience to gene regulation, we monitored the phosphorylation of the zebra finch extracellular signal-regulated kinase (ERK) protein by immunoblotting. Initial presentations of novel song (but not tones or noise) resulted in a rapid increase in ERK phosphorylation, followed by a return to basal levels within 5 min. This response was localized to the auditory forebrain where the zenk gene is activated. Sustained repetition of one song caused a selective habituation of the ERK response: a different song triggered another cycle of ERK phosphorylation without altering the habituated response to the first. To test directly for a role of ERK in experience-dependent zenk gene regulation, we infused an inhibitor of mitogen-activated and extracellular-regulated protein kinase kinase (MEK-1; the enzyme responsible for ERK activation) unilaterally into one auditory lobule just before song stimulation. The song-induced increase in zenk mRNA was blocked on the side of the injection, but not on the contralateral (uninfused) side. These results show that ERK phosphorylation is necessary for the initiation of the zenk gene response to novel song and identify ERK as a plausible site of signal integration underlying the selective habituation of genomic responses to a repeated song.

Sexual preferences for songs in female domestic canaries ( Serinus canaria ): can late song exposure, without social reinforcement, influence the effects of early tutoring?

We studied the relative effects of early and late song exposure without social reinforcement on female sexual preferences in the domestic canary (Serinus canaria). Young female canaries were tape-tutored during their first 4 months of life with songs of either domestic or wild male canaries (DT and WT conditions). When they reached sexual maturity, these females were placed in breeding conditions and some of them were re-exposed to songs. During this “late exposure” the females, according to their experimental group, were either presented with new domestic or wild songs (DL and WL conditions) over 40 days, or were kept without song stimulation (− condition). Afterwards, we assessed the sexual preferences of all the females for domestic or wild songs using the copulation solicitation display assay. The results showed that both DT/– and DT/DL females showed a clear preference for domestic songs. However, whereas WT/WL females preferred wild songs; WT/– females did not show any preference. Finally, DT/WL and WT/DL females failed to show any preference. It appeared that a second song experience at the beginning of their first breeding season, without any social reinforcement, allowed the emergence or stabilisation of early preferences, or interfered with these early preferences depending on whether the song category used during the late exposure phase matched or not the song category used during the early tutoring phase, and also depending on which category was used during the first tutoring phase. This behavioural plasticity could help young adult females to adjust the ‘standard’ they built during infancy to new environmental conditions.

Development of song responses in the zebra finch caudomedial neostriatum: role of genomic and electrophysiological activities.

Zebra finches first form demonstrable memories of specific songs between 25 and 35 days of age--several days after fledging from the nest. What accounts for the late onset of specific song memory formation? Here we investigated physiological development of the caudomedial neostriatum (NCM), part of the avian analogue of auditory cortex and a probable component of the system involved in song perception. Two types of physiological responses were characterized: electrophysiological (single-unit spike rate) and genomic (induction of the immediate early gene zenk, also known as zif-268, egr-1, ngfi-a, krox-24). We found that by day 20, zebra finches already have robust electrophysiological responses in NCM to song stimulation. Spike activity was greater in response to conspecific songs compared to heterospecific songs, white noise, or tones, and approximately 10% of the units showed selective responses to forward versus reversed songs. In contrast, at this age the zenk gene is expressed at a constitutively high level and undergoes no further induction in response to song presentation. At day 30, electrophysiological responses remained similar, but the zenk gene began to shift from a constitutive to an inducible pattern of expression. These results are consistent with a general role for NCM in the representation of song auditory patterns, and with a role for zenk gene expression in governing the efficiency of specific song memory storage at different ages.

Early tutoring and adult reproductive behaviour in female domestic canary ( Serinus canaria )

We studied the effect of early tutoring on the subsequent sexual preferences and reproductive activity of female domestic canaries (Serinus canaria). Young female canaries were exposed during the first 4 months of life to songs of either domestic or wild canaries. When adult, these females were again exposed to domestic or wild songs. In the first experiment, the sexual responses of the females to unfamiliar domestic and wild songs were quantified with the copulation solicitation display (CSD) assay. In the second experiment, the same females were tested again with modified tutoring songs. In the third experiment, song stimulation of nest-building and egg-laying was studied. Domestic-strain-tutored females gave more CSDs to domestic than to wild songs. In contrast, wild-strain-tutored females showed no sexual preference. We propose that the sexual preference of adult domestic-strain-tutored female canaries for domestic songs is the consequence of learning and categorisation processes. The discrepancy between the results of the domestic-strain-tutored females and those of the wild-strain-tutored females suggests that female canaries have a predisposition to learn songs of their own strain rather than songs of an alien strain. In the third experiment nest-building and egg-laying activities appeared to be unaffected by early tutoring conditions: there was no significant differential effect of the different tutoring and exposure conditions on nest-building and egg-laying scores. Mate attraction and stimulation of females’ reproductive activity appear to be two separate functions of male song, which may have been shaped by different evolutionary constraints.

Does sex or photoperiodic condition influence ZENK induction in response to song in European starlings?

Variables such as the photoperiod a bird experiences, or its sex, affect behavioral responses to song. The present experiment investigated whether song-induced expression of the immediate-early gene ZENK is also influenced by sex or photoperiod. We examined the expression of the protein product of ZENK in wild-caught male and female European starlings ( Sturnus vulgaris) in different photoperiodic conditions. In the first experiment, adult reproductively active male and female starlings were presented with either conspecific male song or no song in experimental chambers. In the second experiment, conspecific male song was presented to reproductively active and reproductively inactive adult female starlings. Localization of the ZENK protein product revealed that song stimulation resulted in a significant increase in the number of ZENK immunoreactive (-ir) cells in the caudomedial neostriatum (NCM) and the caudomedial hyperstriatum ventrale (cmHV) compared with unstimulated birds. No differences in the number of ZENK-ir neurons were observed between males and females or between reproductively active and inactive females. Thus, the present data indicate that the number of cells expressing ZENK in NCM and cmHV following song playback does not vary with sex or photoperiod in starlings.

Male Song Stimulation of Female Reproduction in Canaries: Features Relevant to Sexual Displays are not Relevant to Nest‐building or Egg‐laying

AbstractRecent studies conducted in our laboratory have demonstrated that a special type of song phrase (‘sexy’ phrases) containing bipartite syllables composed of abrupt frequency falls and short silences stimulate female canaries to solicit for copulation. The study was undertaken to determine whether sexy phrases also facilitated other aspects of the reproductive activity of the female canary, namely, nest‐building and egg‐laying. During the first experiment, we studied the effect of sexy and non‐sexy songs on copulation solicitation displays in 1‐year‐old females without reproductive experience and in mature females with previous reproductive experience. We confirmed that sexy songs elicited more sexual responses than did non‐sexy songs in yearlings and in mature females. During the second experiment, we studied the effect of male songs on nest‐building activities and egg‐laying in 1‐year‐old inexperienced females, and in mature, experienced females. The songs of conspecific males significantly triggered and increased nest‐building behaviour in female canaries whatever their age or reproductive experience. In contrast, song effects on egg‐laying were only found in young females. One‐year‐old inexperienced females exposed to sexy or non‐sexy songs laid more eggs and laid earlier than did 1‐year‐old inexperienced controls; no such differences were observed in mature, experienced females. The efficiency of songs in promoting nest‐building or egg‐laying appeared to be unrelated to their efficiency in eliciting sexual responses. No difference was found between females exposed to sexy songs and females exposed to non‐sexy songs; differences were only found between control and the two groups of song‐exposed females. This result demands further experiments in order to determine whether other song phrase types may account for the stimulating effects of male song on female nest‐building and egg‐laying.

Song-based species discrimination and behaviour assessment by female blackbirds, Turdus merula

Abstract. The ability of female blackbirds to integrate information about a singer's species and possible ensuing behaviour from different parameters of the introductory part (i.e. the motif part) of the blackbird's song was tested. Females were made receptive to song stimulation by oestradiol injections and tested with tapes with modified motif parts in an anechoic chamber using a broad spectrum of female reproductive behaviour as response measures. Females, like males, attended to the natural upper and/or lower limits of variation in seven parameters in the process of species discrimination: (1) the frequency level and (2) the duration of sound figures (i.e. sounds separated by pauses greater than 50 ms); variation across the motif part in the sound figures' (3) frequency level, (4) duration and (5) amplitude; (6) the ratio of the sound pressure of the fundamentals to that of the first and second overtones; and (7) the relative proportions of the three types of sound figures (i.e. (a) whistles; (b) trills, gurgles and squeaks; and (c) compounds of a and b). The results suggest that females, unlike males, disregard both the upper limit of variation in the frequency level in the process of species discrimination, and the degree of modulation of the sound figures' amplitude functions (i.e. envelopes) and, perhaps, the pauses between the sound figures in the process of assessing the ensuing behaviour of the singer. The validity of such sexual differences in song decoding and possible factors leading to their evolution are discussed. The present differences all concern parameters that degrade more when passing through dense vegetation to females than when passing through the more open communication channels to males. This is consistent with the 'song-degradation' hypothesis that one of the sexes may pay less attention than the other to certain song features if these features are degraded to a different extent on their way to the two sexes.

The Response of Silvereyes to Individual Variation in their Territorial Song

Playback experiments and an investigation of song interaction were used to examine the response of Silvereyes Zosterops lateralis chlorocephala to the individual variation in their territorial song. The Silvereyes did not respond differentially to playback of neighbours' or strangers' songs, nor did they habituate to the presence of playback song within or between experiments. The continuous nature of the individual variation in the Silvereyes' songs did not allow their perception of specific individuals or groups on the basis of song structure. Time of day, the individual's song used and the age of the individual responding did not significantly affect the response. There was no apparent interaction of song between territorial males, either in acoustic avoidance or song stimulation.

Song Parts Adapted to Function Both at Long and Short Ranges May Communicate Information about the Species to Female Blackbirds Turdus merula

Blackbird females were hand-raised in the laboratory and made receptive to song stimulation by long-day conditions and estradiol injections. Playback experiments in an anechoic chamber comparing responses of the females to different conspecific songs and songs of foreign species indicate that females gain information about species identity from the conspecific songs. Not only the complete song seems to have a role in species recognition, but also each of its two components, the long-ranging motif part and the short-ranging twitter part, the latter suggesting an adaptation to a life in dense vegetation.