Song System Research Articles

Induction of an immortalized songbird cell line allows for gene characterization and knockout by CRISPR-Cas9

The zebra finch is one of the most commonly studied songbirds in biology, particularly in genomics, neuroscience and vocal communication. However, this species lacks a robust cell line for molecular biology research and reagent optimization. We generated a cell line, designated CFS414, from zebra finch embryonic fibroblasts using the SV40 large and small T antigens. This cell line demonstrates an improvement over previous songbird cell lines through continuous and density-independent growth, allowing for indefinite culture and monoclonal line derivation. Cytogenetic, genomic, and transcriptomic profiling established the provenance of this cell line and identified the expression of genes relevant to ongoing songbird research. Using this cell line, we disrupted endogenous gene sequences using S.aureus Cas9 and confirmed a stress-dependent localization response of a song system specialized gene, SAP30L. The utility of CFS414 cells enhances the comprehensive molecular potential of the zebra finch and validates cell immortalization strategies in a songbird species.

Research on the Architecture of Digital Song and Dance Costume Design System Based on Intelligent Deep Learning Algorithm

In order to improve the intelligence and reliability of song and dance costume design, this article combines the intelligent deep learning algorithm to improve the digital costume design process. In order to solve the problem of minimizing the energy of image deformation, this paper proposes to transform it into a sparse matrix to solve the problem, which is solved by constructing a coefficient matrix and multiple iterations. After the algorithm is improved, this article combines the digital intelligence method to construct the digital song and dance costume design system structure based on the intelligent deep learning algorithm and analyzes its work flow. Finally, this paper verifies the method in this paper through feasibility analysis and system performance test. From the experimental results, it can be seen that the digital song and dance costume design system based on the intelligent deep learning algorithm proposed in this article has a good effect in the song and dance costume design.

Ancient Beliefs and Traditional Singing of Ukrainians: Gender Differentiation

Gender differentiation in the Ukrainian song tradition is most consistently manifested in the ritual system, and partly – in the epic tradition and lyrical song tradition. The primary attention in the article is paid to the ritual sphere, first of all to the calendar cycle. The paper highlights in detail the traditional distribution of functions between men and women in pre-Christian rituals, during which, according to ancient ideas and beliefs, there was some contact between «that» (sacred) and «this» («profane») worlds.
 Males, or more precisely, boys who were members of the so-called «parubotchi gromady» (young men communities) took an active part in the rites only once a year – at the beginning of the calendar-time cycle that is, in winter (in ancient times this happened in the spring) during the rituals of the yards circumambulation. The main purpose of these actions was to wish good for each family from the dead ancestors for the coming year (verbal magic), and in return, the ancestors received gifts – sacrificial food from representatives of the living world to appease them for the next year.
 Women, as representatives of «this» world, maintained contact with otherworldly forces throughout the entire agrarian period from sowing to harvest, as well as in ceremonies associated with the birth of a child, a wedding, or escorting the deceased to the afterlife. In times of crisis in the development of nature and human life, they turned to their deceased ancestors for help. The magical instrument of this connection was the voice, which filled the ritual texts with specific ritual timbre-intonation.
 The gender distribution in other genres of Ukrainian traditional song is somewhat different. Thus, if in the epic songs the prerogative belongs to men, then the lyric song system is characterized by the joint and almost equal participation of men and women. However, it should be noted that the performers of social songs were predominantly men, and women sang family lyric songs. But the most common was a mixed lineup of singing groups. Even more this property is inherent in the late layer of lyrical song performance.
 The author also draws attention to the age aspect of the performance of ritual and non-ritual songs in the Ukrainian tradition

The singing question: re-conceptualizing birdsong.

Birdsong has been the subject of broad research from a variety of sub-disciplines and has taught us much about the evolution, function, and mechanisms driving animal communication and cognition. Typically, birdsong refers to the specialized vocalizations produced by oscines. Historically, much of the research on birdsong was conducted in north temperate regions (specifically in Europe and North America) leading to multiple biases. Due to these historic biases these vocalizations are generally considered to be highly sexually dimorphic, heavily shaped by sexual selection and essential for courtship and territoriality. Song is also typically defined as a learned trait shaped by cultural evolution. Together, this framework focuses research specifically on males, particularly during the north temperate breeding season - reflecting and thereby reinforcing this framework. The physiological underpinnings of song often emphasize the role of the hypothalamic-pituitary-gonadal axis (associated with breeding changes) and the song control system (underlying vocal learning). Over the years there has been great debate over which features of song are essential to the definition of birdsong, which features apply broadly to contexts outside males in the north temperate region, and over the importance of having a definition at all. Importantly, the definitions we use can both guide and limit the progress of research. Here, we describe the history of these definitions, and how these definitions have directed and restricted research to focus on male song in sexually selected contexts. Additionally, we highlight the gaps in our scientific knowledge, especially with respect to the function and physiological mechanisms underlying song in females and in winter, as well as in non-seasonally breeding species. Furthermore, we highlight the problems with using complexity and learning as dichotomous variables to categorize songs and calls. Across species, no one characteristic of song - sexual dimorphism, seasonality, complexity, sexual selection, learning - consistently delineates song from other songbird vocal communication. We provide recommendations for next steps to build an inclusive information framework that will allow researchers to explore nuances in animal communication and promote comparative research. Specifically, we recommend that researchers should operationalize the axis of variation most relevant to their study/species by identifying their specific question and the variable(s) of focus (e.g. seasonality). Researchers should also identify the axis (axes) of variation (e.g. degree of control by testosterone) most relevant to their study and use language consistent with the question and axis (axes) of variation (e.g. control by testosterone in the seasonal vocal production of birds).

Unilateral vocal nerve resection alters neurogenesis in the avian song system in a region-specific manner.

New neurons born in the adult brain undergo a critical period soon after migration to their site of incorporation. During this time, the behavior of the animal may influence the survival or culling of these cells. In the songbird song system, earlier work suggested that adult-born neurons may be retained in the song motor pathway nucleus HVC with respect to motor progression toward a target song during juvenile song learning, seasonal song restructuring, and experimentally manipulated song variability. However, it is not known whether the quality of song per se, without progressive improvement, may also influence new neuron survival. To test this idea, we experimentally altered song acoustic structure by unilateral denervation of the syrinx, causing a poor quality song. We found no effect of aberrant song on numbers of new neurons in HVC, suggesting that song quality does not influence new neuron culling in this region. However, aberrant song resulted in the loss of left-side dominance in new neurons in the auditory region caudomedial nidopallium (NCM), and a bilateral decrease in new neurons in the basal ganglia nucleus Area X. Thus new neuron culling may be influenced by behavioral feedback in accordance with the function of new neurons within that region. We propose that studying the effects of singing behaviors on new neurons across multiple brain regions that differentially subserve singing may give rise to general rules underlying the regulation of new neuron survival across taxa and brain regions more broadly.

Perineuronal nets in HVC and plasticity in male canary song.

Songbirds learn their vocalizations during developmental sensitive periods of song memorization and sensorimotor learning. Some seasonal songbirds, called open-ended learners, recapitulate transitions from sensorimotor learning and song crystallization on a seasonal basis during adulthood. In adult male canaries, sensorimotor learning occurs each year in autumn and leads to modifications of the syllable repertoire during successive breeding seasons. We previously showed that perineuronal nets (PNN) expression in song control nuclei decreases during this sensorimotor learning period. Here we explored the causal link between PNN expression in adult canaries and song modification by enzymatically degrading PNN in HVC, a key song control system nucleus. Three independent experiments identified limited effects of the PNN degradation in HVC on the song structure of male canaries. They clearly establish that presence of PNN in HVC is not required to maintain general features of crystallized song. Some suggestion was collected that PNN are implicated in the stability of song repertoires but this evidence is too preliminary to draw firm conclusions and additional investigations should consider producing PNN degradations at specified time points of the seasonal cycle. It also remains possible that once song has been crystallized at the beginning of the first breeding season, PNN no longer play a key role in determining song structure; this could be tested by treatments with chondroitinase ABC at key steps in ontogeny. It would in this context be important to develop multiple stereotaxic procedures allowing the simultaneous bilateral degradation of PNN in several song control nuclei for extended periods.

Evidence That Artificial Light at Night Induces Structure-Specific Changes in Brain Plasticity in a Diurnal Bird.

We recently reported that artificial light at night (ALAN), at ecologically relevant intensities (1.5, 5 lux), increases cell proliferation in the ventricular zone and recruitment of new neurons in several forebrain regions of female zebra finches (Taeniopygia guttata), along with a decrease of total neuronal densities in some of these regions (indicating possible neuronal death). In the present study, we exposed male zebra finches to the same ALAN intensities, treated them with 5′-bromo-2′-deoxyuridine, quantified cell proliferation and neuronal recruitment in several forebrain regions, and compared them to controls that were kept under dark nights. ALAN increased cell proliferation in the ventricular zone, similar to our previous findings in females. We also found, for the first time, that ALAN increased new neuronal recruitment in HVC and Area X, which are part of the song system in the brain and are male-specific. In other brain regions, such as the medial striatum, nidopallium caudale, and hippocampus, we recorded an increased neuronal recruitment only in the medial striatum (unlike our previous findings in females), and relative to the controls this increase was less prominent than in females. Moreover, the effect of ALAN duration on total neuronal densities in the studied regions varied between the sexes, supporting the suggestion that males are more resilient to ALAN than females. Suppression of nocturnal melatonin levels after ALAN exhibited a light intensity-dependent decrease in males in contrast to females, another indication that males might be less affected by ALAN. Taken together, our study emphasizes the importance of studying both sexes when considering ALAN effects on brain plasticity.

Synthetic Birdsongs as a Tool to Induce, and Iisten to, Replay Activity in Sleeping Birds.

Birdsong is a complex vocal behavior, which emerges out of the interaction between a nervous system and a highly nonlinear vocal device, the syrinx. In this work we discuss how low dimensional dynamical systems, interpretable in terms of the biomechanics involved, are capable of synthesizing realistic songs. We review the experimental and conceptual steps that lead to the formulation of low dimensional dynamical systems for the song system and describe the tests that quantify their success. In particular, we show how to evaluate computational models by comparing the responses of highly selective neurons to the bird’s own song and to synthetic copies generated mathematically. Beyond testing the hypothesis behind the model’s construction, these low dimensional models allow designing precise stimuli in order to explore the sensorimotor integration of acoustic signals.

One-to-one innervation of vocal muscles allows precise control of birdsong

The motor control resolution of any animal behavior is limited to the minimal force step available when activating muscles, which is set by the number and size distribution of motor units (MUs) and muscle-specific force. Birdsong is an excellent model system for understanding acquisition and maintenance of complex fine motor skills, but we know surprisingly little about how the motor pool controlling the syrinx is organized and how MU recruitment drives changes in vocal output. Here we developed an experimental paradigm to measure MU size distribution using spatiotemporal imaging of intracellular calcium concentration in cross-sections of living intact syrinx muscles. We combined these measurements with muscle stress and an invitro syrinx preparation to determine the control resolution of fundamental frequency (fo), a key vocal parameter, in zebra finches. We show that syringeal muscles have extremely small MUs, with 40%-50% innervating ≤3 and 13%-17% innervating a single muscle fiber. Combined with the lowest specific stress (5mN/mm2) known to skeletal vertebrate muscle, small force steps by the major fo controlling muscle provide control of 50-mHz to 7.3-Hz steps per MU. We show that the song system has the highest motor control resolution possible in the vertebrate nervous system and suggest this evolved due to strong selection on fine gradation of vocal output. Furthermore, we propose that high-resolution motor control was a key feature contributing to the radiation of songbirds that allowed diversification of song and speciation by vocal space expansion.

The importance of Vasanthan Kooththu (Art Form) songs in revealing the existence of human social and professional life

The culture of a community plays a major role in narrating the history and life of that particular community. This culture includes various aspects such as religion, ritual, faith, food, customs or practices as well as art and culture. Among these aspects, art is not only for entertainment; but it is also for exposing the social character and tradition of that particular community. The undeniable thing here is the factor that art forms in every country, and in every region are not only to create entertainment but also to preserve the antiquity of their existence up to now. In that respect, Vasanthan Kooththu, one of the Sri Lankan Tamil art forms, is a notable art form in that category. Even though this art form is found in Batticaloa and Jaffna areas, they have differences between each other. This study focuses on the songs of Vasanthan Kooththu performed in the Katuthavalai area of ​​Batticaloa. This type of Kooththu is an art form of tapping and dancing with two sticks in the hands; however, these sticks are also used to express the function of the meaning of the song with dance. Even though this type of Kooththu is a dance form, the greatness of the songs used in this Kooththu is significant. There are 62 types of Vasanthan poems have been in use in the Art Form performances. These have been compiled around the year 1940. These 62 genres of songs are divided into six genres and compiled, namely 'Kattiyam', 'Thoththiram', 'Sariththiram', 'Tholil', 'Vedikkai', 'Vilaiyaattu' and are still in the practice during Kooththu performance. As mentioned earlier, these Kooththu song systems emphasize the art expression of the culture of a community. The songs related to the professions or job involved in these Kooththu songs express the whole series of activities of the agricultural industry. The reason that these Kooththu songs to aim to explore only a specific industry is to be explored. It is vital to discover this factor; and this article explores about how these Kooththu songs are still in operation today as a popular form of folk music performance to describe a particular social professional life, beyond globalization trends, imposition and blends of colonization thinking. This article also explores the significance of these Kooththu songs hold, the importance they have gained in the life of people as well as the value that these Kooththu songs have even today.

Syringeal vocal folds do not have a voice in zebra finch vocal development

Vocal behavior can be dramatically changed by both neural circuit development and postnatal maturation of the body. During song learning in songbirds, both the song system and syringeal muscles are functionally changing, but it is unknown if maturation of sound generators within the syrinx contributes to vocal development. Here we densely sample the respiratory pressure control space of the zebra finch syrinx in vitro. We show that the syrinx produces sound very efficiently and that key acoustic parameters, minimal fundamental frequency, entropy and source level, do not change over development in both sexes. Thus, our data suggest that the observed acoustic changes in vocal development must be attributed to changes in the motor control pathway, from song system circuitry to muscle force, and not by material property changes in the avian analog of the vocal folds. We propose that in songbirds, muscle use and training driven by the sexually dimorphic song system are the crucial drivers that lead to sexual dimorphism of the syringeal skeleton and musculature. The size and properties of the instrument are thus not changing, while its player is.

Song learning and plasticity in songbirds.

Birdsong provides a fascinating system to study both behavioral and neural plasticity. Oscine songbirds learn to sing, exhibiting behavioral plasticity both during and after the song-learning process. As a bird learns, its song progresses from a plastic and highly variable vocalization into a more stereotyped, crystallized song. However, even after crystallization, song plasticity can occur: some species' songs become more stereotyped over time, whereas other species can incorporate new song elements. Alongside the changes in song, songbirds' brains are also plastic. Both song and neural connections change with the seasons in many species, and new neurons can be added to the song system throughout life. In this review, we highlight important research on behavioral and neural plasticity at multiple timescales, from song development in juveniles to lifelong modifications of learned song.

Intrinsic plasticity and birdsong learning

Although information processing and storage in the brain is thought to be primarily orchestrated by synaptic plasticity, other neural mechanisms such as intrinsic plasticity are available. While a number of recent studies have described the plasticity of intrinsic excitability in several types of neurons, the significance of non-synaptic mechanisms in memory and learning remains elusive. After reviewing plasticity of intrinsic excitation in relation to learning and homeostatic mechanisms, we focus on the intrinsic properties of a class of basal-ganglia projecting song system neurons in zebra finch, how these related to each bird’s unique learned song, how these properties change over development, and how they are maintained dynamically to rapidly change in response to auditory feedback perturbations. We place these results in the broader theme of learning and changes in intrinsic properties, emphasizing the computational implications of this form of plasticity, which are distinct from synaptic plasticity. The results suggest that exploring reciprocal interactions between intrinsic and network properties will be a fruitful avenue for understanding mechanisms of birdsong learning.

Neural Code of Motor Planning and Execution during Goal-Directed Movements in Crows.

The planning and execution of head-beak movements are vital components of bird behavior. They require integration of sensory input and internal processes with goal-directed motor output. Despite its relevance, the neurophysiological mechanisms underlying action planning and execution outside of the song system are largely unknown. We recorded single-neuron activity from the associative endbrain area nidopallium caudolaterale (NCL) of two male carrion crows (Corvus corone) trained to plan and execute head-beak movements in a spatial delayed response task. The crows were instructed to plan an impending movement toward one of eight possible targets on the left or right side of a touchscreen. In a fraction of trials, the crows were prompted to plan a movement toward a self-chosen target. NCL neurons signaled the impending motion direction in instructed trials. Tuned neuronal activity during motor planning categorically represented the target side, but also specific target locations. As a marker of intentional movement preparation, neuronal activity reliably predicted both target side and specific target location when the crows were free to select a target. In addition, NCL neurons were tuned to specific target locations during movement execution. A subset of neurons was tuned during both planning and execution period; these neurons experienced a sharpening of spatial tuning with the transition from planning to execution. These results show that the avian NCL not only represents high-level sensory and cognitive task components, but also transforms behaviorally-relevant information into dynamic action plans and motor execution during the volitional perception-action cycle of birds.SIGNIFICANCE STATEMENT Corvid songbirds have become exciting new models for understanding complex cognitive behavior. As a key neural underpinning, the endbrain area nidopallium caudolaterale (NCL) represents sensory and memory-related task components. How such representations are converted into goal-directed motor output remained unknown. In crows, we report that NCL neurons are involved in the planning and execution of goal-directed movements. NCL neurons prospectively signaled motion directions in instructed trials, but also when the crows were free to choose a target. NCL neurons showed a target-specific sharpening of tuning with the transition from the planning to the execution period. Thus, the avian NCL not only represents high-level sensory and cognitive task components, but also transforms relevant information into action plans and motor execution.

The cells of songbird motor circuits

Neuroscience Birds have complex motor and cognitive abilities that rival or exceed the performance of many mammals, but their brains are organized in a notably different way. Parts of the bird brain have been functionally compared to the mammalian neocortex. However, it is still controversial to what extent these regions are truly homologous with the neocortex or if instead they are examples of evolutionary convergence. Colquitt et al. used single-cell sequencing to identify and characterize the major classes of neurons that comprise the song-control system in birds (see the Perspective by Tosches). They found multiple previously unknown neural classes in the bird telencephalon and shed new light on the long-standing controversy regarding the nature of homology between avian and mammalian brains. Science , this issue p. [eabd9704][1]; see also p. [676][2] [1]: /lookup/doi/10.1126/science.abd9704 [2]: /lookup/doi/10.1126/science.abf9551

A histological study of the song system of the carrion crow (Corvus corone).

The song system of songbirds (oscines) is one of the best studied neuroethological model systems. So far, it has been treated as a relatively constrained sensorimotor system. Songbirds such as crows, however, are also known for their capability to cognitively control their audio-vocal system. Yet, the neuroanatomy of the corvid song system has never been explored systematically. We aim to close this scientific gap by presenting a stereotactic investigation of the extended song system of the carrion crow (Corvus corone), an oscine songbird of the corvid family that has become an interesting model system for cognitive neuroscience. In order to identify and delineate the song nuclei, the ascending auditory nuclei, and the descending vocal-motor nuclei, four stains were applied. In addition to the classical Nissl-, myelin-, and a combination of Nissl-and-myelin staining, staining for tyrosine hydroxylase was used to reveal the distribution of catecholaminergic neurons (dopaminergic, noradrenergic, and adrenergic) in the song system. We show that the crow brain contains the important song-related nuclei, including auditory input and motor output structures, and map them throughout the brain. Fiber-stained sections reveal putative connection patterns between the crow's song nuclei comparable to other songbirds.

Estrogen and sex-dependent loss of the vocal learning system in female zebra finches

Sex hormones alter the organization of the brain during early development and coordinate various behaviors throughout life. In zebra finches, song learning is limited to males, with the associated song learning brain pathways only maturing in males and atrophying in females. While this atrophy can be prevented by treating females with exogenous estrogen during early post-hatch development, the requirement of estrogen during normal male song system development is uncertain. For the first time in songbirds, we administered exemestane, a potent third generation estrogen synthesis inhibitor, from the day of hatching until adulthood in order to reassess the role of estrogen in song circuit development. We examined the behavior, brain anatomy, and transcriptomes of individual song nuclei in these pharmacologically manipulated animals. We found that males with long-term exemestane treatment had diminished male-specific plumage and impaired song learning, but minimal effect on song nuclei sizes and their specialized transcriptome. Consistent with prior findings, females with long-term estrogen treatment retained a functional song system with song nuclei that had specialized gene expression similar, but not identical to males. We also observed that different song nuclei responded to estrogen manipulation differently, with Area X in the striatum being the most altered by estrogen modulation. These findings support the hypothesis that song learning is an ancestral trait in both sexes that was subsequently suppressed in females of some species and that estrogen has come to play a critical role in modulating this suppression as well as refinement of song learning.

Central prolactin receptor distribution and pSTAT5 activation patterns in breeding and non-breeding zebra finches (Taeniopygia guttata)

The hormone prolactin has many diverse functions across taxa such as osmoregulation, metabolism, and reproductive behavior. In ring doves, central prolactin action is important for parental care and feeding behavior. However, there is a considerable lack of information on the distribution of the prolactin receptor (PRLR) in the avian CNS to test the hypothesis that prolactin mediates these and other functions in other birds. In order to advance this research, we collected brains from breeding and non-breeding zebra finches to map the PRLR distribution using immunohistochemistry. We found PRLRs are distributed widely across the brain, both in hypothalamic sites known to regulate parental care and feeding, but also in many non-hypothalamic sites, including the tectofugal visual pathway, song system regions, reward associated areas, and pallium. This raises the possibility that prolactin has other functions throughout the brain that are not necessarily related to feeding or parental care. In addition, we also stained brains for pSTAT5, a transcription factor which is expressed when the PRLR is activated and is used as a marker for PRLR activity. We found several notable differences in pSTAT5 activity due to the breeding state of the animal, in both directions, further supporting the hypothesis that prolactin has many diverse functions in the brain both within and outside times of breeding. Together, this study represents the first essential step to inform the design of causative studies which manipulate PRLR-expressing cells to test their role in a wide variety of behaviors and other physiological functions.

Differential development of myelin in zebra finch song nuclei.

Songbirds learn vocalizations by hearing and practicing songs. As song develops, the tempo becomes faster and more precise. In the songbird brain, discrete nuclei form interconnected myelinated circuits that control song acquisition and production. The myelin sheath increases the speed of action potential propagation by insulating the axons of neurons and by reducing membrane capacitance. As the brain develops, myelin increases in density, but the time course of myelin development across discrete song nuclei has not been systematically studied in a quantitative fashion. We tested the hypothesis that myelination develops differentially across time and song nuclei. We examined myelin development in the brains of the zebra finch (Taeniopygia guttata) from chick at posthatch day (d) 8 to adult (up to 147 d) in five major song nuclei: HVC (proper name), robust nucleus of the arcopallium (RA), Area X, lateral magnocellular nucleus of the anterior nidopallium, and medial portion of the dorsolateral thalamic nucleus (DLM). All of these nuclei showed an increase in the density of myelination during development but at different rates and to different final degrees. Exponential curve fits revealed that DLM showed earlier myelination than other nuclei, and HVC showed the slowest myelination of song nuclei. Together, these data show differential maturation of myelination in different portions of the song system. Such differential maturation would be well placed to play a role in regulating the development of learned song.

Sex-and Region-Dependent Expression of the Autism-Linked ADNP Correlates with Social- and Speech-Related Genes in the Canary Brain

The activity-dependent neuroprotective protein (ADNP) syndrome is an autistic-like disorder, instigated by mutations in ADNP. This syndrome is characterized by developmental delays, impairments in speech, motor function, abnormal hearing, and intellectual disabilities. In the Adnp-haploinsufficient mouse model, many of these impediments are evident, appearing in a sex-dependent manner. In zebra finch songbird (ZF; Taeniopygia guttata), an animal model used for song/language studies, ADNP mRNA most robust expression is observed in the cerebrum of young males, potentially corroborating with male ZF exclusive singing behavior and developed cerebral song system. Herein, we report a similar sex-dependent ADNP expression profile, with the highest expression in the cerebrum (qRT-PCR) in the brain of another songbird, the domesticated canary (Serinus canaria domestica). Additional analyses for the mRNA transcripts of the ADNP regulator, vasoactive intestinal peptide (VIP), sister gene ADNP2, and speech-related Forkhead box protein P2 (FoxP2) revealed multiple sex and brain region-dependent positive correlations between the genes (including ADNP). Parallel transcript expression patterns for FoxP2 and VIP were observed alongside specific FoxP2 increase in males compared with females as well as VIP/ADNP2 correlations. In spatial view, a sexually independent extensive form of expression was found for ADNP in the canary cerebrum (RNA in situ hybridization). The songbird cerebral mesopallium area stood out as a potentially high-expressing ADNP tissue, further strengthening the association of ADNP with sense integration and auditory memory formation, previously implicated in mouse and human.