Functional specialization and distributed processing across marmoset lateral prefrontal subregions.

A prominent aspect of primate lateral prefrontal cortex organization is its division into several cytoarchitecturally distinct subregions. Neurophysiological investigations in macaques have provided evidence for the functional specialization of these subregions, but an understanding of the relative representational topography of sensory, social, and cognitive processes within them remains elusive. One explanatory factor is that evidence for functional specialization has been compiled largely from a patchwork of findings across studies, in many animals, and with considerable variation in stimulus sets and tasks. Here, we addressed this by leveraging the common marmoset (Callithrix jacchus) to carry out large-scale neurophysiological mapping of the lateral prefrontal cortex using high-density microelectrode arrays, and a diverse suite of test stimuli including faces, marmoset calls, and spatial working memory task. Task-modulated units and units responsive to visual and auditory stimuli were distributed throughout the lateral prefrontal cortex, while those with saccade-related activity or face-selective responses were restricted to 8aV, 8aD, 10, 46 V, and 47. Neurons with contralateral visual receptive fields were limited to areas 8aV and 8aD. These data reveal a mixed pattern of functional specialization in the lateral prefrontal cortex, in which responses to some stimuli and tasks are distributed broadly across lateral prefrontal cortex subregions, while others are more limited in their representation.

A cross-species framework for classifying sound-movement couplings.

Sound and movement are entangled in animal communication. This is obviously true in the case of sound-constituting vibratory movements of biological structures which generate acoustic waves. A little less obvious is that other moving structures produce the energy required to sustain these vibrations. In many species, the respiratory system moves to generate the expiratory flow which powers the sound-constituting movements (sound-powering movements). The sound may acquire additional structure via upper tract movements, such as articulatory movements or head raising (sound-filtering movements). Some movements are not necessary for sound production, but when produced, impinge on the sound-producing process due to weak biomechanical coupling with body parts (e.g., respiratory system) that are necessary for sound production (sound-impinging movements). Animals also produce sounds contingent with movement, requiring neuro-physiological control regimes allowing to flexibly couple movements to a produced sound, or coupling movements to a perceived external sound (sound-contingent movement). Here, we compare and classify the variety of ways sound and movements are coupled in animal communication; our proposed framework should help structure previous and future studies on this topic.

Bison mother-offspring acoustic communication.

Mother-offspring communication is especially crucial for social species in order to synchronize activities essential for early survival including nursing, resting, maintaining proximity during group movements between food or water sources, and locating one another if separated in a large social group. One of the most social ungulate species in North America is the American Bison (Bison bison), formerly known as buffalo. Adult female bison associate with their young for over a year and communication between mother and offspring is likely essential for establishing and maintaining a bond upon which the life of a calf depends. One goal of this study was to quantify and compare the acoustic form of vocalizations of adult female, subadult, and calf bison and to determine how age classes differed in call structure. The other goal was to identify the contexts in which bison vocalized. Vocalizations of 101 bison (53 adult females, 15 subadults, 33 calves) in a semi-free-ranging herd in Montana were analyzed and found to be pulsatile sounds, unlike vocalizations of bison bulls or domestic cows and calves. Vocalizations of bison cows, subadults, and calves differed significantly in total duration, numbers of pulses, pulse duration, and pulse rate. Seven distinct call contexts were identified. The majority of calls were "moving-on calls" (39%), when a cow called and her calf ran to her side and the 2 moved on together, and "contact calls" (21%) when a cow called and her calf called back but neither changed their location. "Imprinting calls" and "nursing calls" were also identified. Mother-offspring acoustic communication in bison appears especially critical for coordinating movements. Understanding the role of acoustic communication in maintaining the bond between bison mothers and their offspring can contribute to the humane management and welfare of this iconic species.

La comunicación madre-cría es especialmente crucial para especies sociales, debido a que permite sincronizar actividades esenciales para la supervivencia temprana, como la lactancia, el descanso, el mantenimiento de proximidad durante los movimientos de los grupo entre las fuentes de alimento o agua y la localización mutua en caso de separación dentro de un grupo social grande. Una de las especies de ungulados más sociales de América del Norte es el bisonte (Bison bison). Las hembras adultas de bisonte se asocian con sus crías durante más de un año y la comunicación entre madre el becerro es probablemente esencial para establecer y mantener un vínculo en el que depende la vida de la cría. Uno de los objetivos de este estudio fue cuantificar y comparar la forma acústica de las vocalizaciones de hembras adultas, subadultas y crías de bisonte, y determinar cómo diferían las clases por edad en la estructura de las llamadas. Otro objetivo fue identificar los contextos en los que se emitían las vocalizaciones. Se analizaron las vocalizaciones de 101 bisontes (53 hembras adultas, 15 hembras subadultas, 33 crías) en un rebaño semi-libre en Montana. Se encontró que estas vocalizaciones eran sonidos pulsátiles, completamente diferentes a los emitidos por los bisontes machos adultos o las vacas y becerros domésticos. Las vocalizaciones diferían significativamente entre las tres clases de edad en su duración total, número de pulsos, duración de los pulsos y ritmo de los pulsos. La mayoría de las llamadas se dieron en dos contextos: "llamadas de avance" (39%), cuando una hembra adulta llamaba y su cría corría a su lado y ambas avanzaban juntas, y "llamadas de contacto" (21%), cuando una hembra adulta llamaba y su cría respondía, pero ninguna cambiaba su ubicación. También se identificaron "llamadas de impronta" y "llamadas de amamantamiento," así como otros tres contextos de llamada. La comunicación acústica madre-cría en bisontes parece especialmente crítica para coordinar los movimientos. Entender el papel de la comunicación acústica en el mantenimiento del vínculo entre las madres y sus crías puede contribuir al manejo humanitario y al bienestar de esta especie icónica. Este trabajo representa el primer estudio que investiga cuantitativamente las señales acústicas de hembras adultas, subadultas y crías de bisontes Norte Americanos mientras se desplazan en condiciones de semi-libertad.

Comparative Study of the Anxiolytic Activity of Songorine in Elevated Plus Maze Test and by Recording Ultrasonic Vocalizations.

It was found that the diterpene alkaloid songorine administered per os to mice at a dose of 25 µg/kg provides a pronounced anxiolytic effect during elevated plus maze testing comparable to the effect of the benzodiazepine anxiolytic phenazepam. Recording of ultrasonic vocalizations of animals revealed an increase in the number of short high-frequency (50 kHz) signals under the action of songorine and the reference drug, which confirms their anti-anxiety properties.

Role of auditory feedback for vocal production learning in the Egyptian fruit bat.

Some species have evolved the ability to use the sense of hearing to modify existing vocalizations, or even create new ones, which enlarges their repertoires and results in complex communication systems.1 This ability corresponds to various forms of vocal production learning that are all possessed by humans and independently displayed by distantly related vertebrates.1,2,3,4,5,6,7 Among mammals, a few species, including the Egyptian fruit bat,8,9,10 would possess such vocal production learning abilities.7 Yet the necessity of an intact auditory system for the development of the Egyptian fruit bat typical vocal repertoire has not been tested. Furthermore, a systematic causal examination of learned and innate aspects of the entire repertoire has never been performed in any vocal learner. Here we addressed these gaps by eliminating pups' sense of hearing at birth and assessing its effects on vocal production in adulthood. The deafening treatment enabled us to both causally test these bats' vocal learning ability and discern learned from innate aspects of their vocalizations. Leveraging wireless individual audio recordings from freely interacting adults, we show that a subset of the Egyptian fruit bat vocal repertoire necessitates auditory feedback. Intriguingly, these affected vocalizations belong to different acoustic groups in the vocal repertoire of males and females. These findings open the possibilities for targeted studies of the mammalian neural circuits that enable sexually dimorphic forms of vocal learning.

The ultrasonic vocalization (USV) syllable profile during neonatal opioid withdrawal and a kappa opioid receptor component to increased USV emissions in female mice.

RATIONALE: Opioid use during pregnancy can lead to negative infant health outcomes, including neonatal opioid withdrawal syndrome (NOWS). NOWS comprises gastrointestinal, autonomic nervous system, and neurological dysfunction that manifest during spontaneous withdrawal. Variability in NOWS severity necessitates a more individualized treatment approach. Ultrasonic vocalizations (USVs) in neonatal mice are emitted in isolation as a stress response and are increased during opioid withdrawal, thus modeling a negative affective state that can be utilized to test new treatments. OBJECTIVES: We sought to identify the behavioral and USV profile, brainstem transcriptomic adaptations, and role of kappa opioid receptors in USVs during neonatal opioid withdrawal. METHODS: We employed a third trimester-approximate opioid exposure model, where neonatal inbred FVB/NJ pups were injected twice-daily with morphine (10mg/kg, s.c.) or saline (0.9%, 20 ul/g, s.c.) from postnatal day(P) 1 to P14. This protocol induces reduced weight gain, hypothermia, thermal hyperalgesia, and increased USVs during spontaneous morphine withdrawal. RESULTS: On P14, there were increased USV emissions and altered USV syllables during withdrawal, including an increase in Complex 3 syllables in FVB/NJ females (but not males). Brainstem bulk mRNA sequencing revealed an upregulation of the kappa opioid receptor (Oprk1), which contributes to withdrawal-induced dysphoria. The kappa opioid receptor (KOR) antagonist, nor-BNI (30 mg/kg, s.c.), significantly reduced USVs in FVB/NJ females, but not males during spontaneous morphine withdrawal. Furthermore, the KOR agonist, U50,488h (0.625 mg/kg, s.c.), was sufficient to increase USVs on P10 (both sexes) and P14 (females only) in FVB/NJ mice. CONCLUSIONS: We identified an elevated USV syllable, Complex 3, and a female-specific recruitment of the dynorphin/KOR system in increased USVs associated with neonatal opioid withdrawal severity.

Autistic-relevant behavioral phenotypes of a mouse model of cyclin-dependent kinase-like 5 deficiency disorder.

Cyclin-dependent kinase-like 5 (CDKL5) deficiency disorder (CDD) is a neurodevelopmental disease caused by mutations in the X-linked CDKL5 gene and characterized by early-onset epilepsy, intellectual disability, and autistic features. To date, the etiological mechanisms underlying CDD are largely unknown and no effective therapies are available. The Cdkl5 knock-out (KO) mouse has been broadly employed in preclinical studies on CDD; Cdkl5-KO mice display neurobehavioral abnormalities recapitulating most CDD symptoms, including alterations in motor, sensory, cognitive, and social abilities. However, most available preclinical studies have been carried out on adult Cdkl5-KO mice, so little is known about the phenotypic characteristics of this model earlier during development. Furthermore, major autistic-relevant phenotypes, for example, social and communication deficits, have been poorly investigated and mostly in male mutants. Here, we assessed the autistic-relevant behavioral phenotypes of Cdkl5-KO mice during the first three post-natal weeks and in adulthood. Males and females were tested, the latter including both heterozygous and homozygous mutants. Cdkl5 mutant pups showed qualitative and quantitative alterations in ultrasonic communication, detected first at 2 weeks of age and confirmed later in adulthood. Increased levels of anxiety-like behaviors were observed in mutants at 3 weeks and in adulthood, when stereotypies, reduced social interaction and memory deficits were also observed. These behavioral effects of the mutation were evident in both sexes, being more marked and varied in homozygous than heterozygous females. These findings provide novel evidence for the autistic-relevant behavioral profile of the Cdkl5 mouse model, thus supporting its use in future preclinical studies investigating CDD pathology and autism spectrum disorders.

Altered vocal communication in adult vasopressin-deficient Brattleboro rats.

The neuropeptide, arginine vasopressin (AVP), has been implicated in social communication across a diverse array of species. Many rodents communicate basic behavioral states with negative versus positive valence through high-pitched vocalizations above the human hearing range (ultrasonic vocalizations; USVs). Previous studies have found that Brattleboro (Bratt) rats, which have a mutation in the Avp gene, exhibit deficits in their USVs from the early postnatal period through adolescence, but the magnitude of this effect appears to decrease from the juvenile to adolescent phase. The present study tested whether Bratt rats continue to exhibit USV deficits in adulthood. USVs of adult male and female Bratt and wild type (WT) rats were recorded in two contexts: a novel environment (empty arena) and a social context (arena filled with bedding soiled by same-sex conspecifics). The number, frequency, and duration of 50 kHz USVs were quantified by DeepSqueak after validation with manual scoring. Twenty-two kHz measures were quantified by manual scoring because DeepSqueak failed to accurately detect USVs in this frequency range. Adult Bratt rats did not exhibit deficits in the number of 50 kHz USVs: male Bratt rats emitted similar 50 kHz USVs as male WT rats, whereas female Bratt rats emitted more USVs than female WT rats. USV frequency and duration were altered in adult Bratt rats, but in a context-dependent manner. Twenty-two kHz USVs were less affected by the Bratt mutation. The present study demonstrates how chronic AVP deficiency impacts social communication across the lifespan. The present findings reveal a complex role for AVP in vocal communication, whereby disruption to the Avp gene leads to sex-, context-, and developmental phase-specific effects on the quantity and spectrotemporal characteristics of rat USVs.

Seasonal variation in the ultrasonic vocal activity of Humboldt's flying squirrel (Glaucomys oregonensis).

Most mammals rely on vocal communication to increase survival and reproductive success. While the functions of audible vocalizations have been well-studied across mammal species, ultrasonic vocalizations in small mammals outside of bats are less understood. North American flying squirrel species (Glaucomys spp.), including the newly described Humboldt's flying squirrel (Glaucomys oregonensis), produce numerous call-types that extend into the ultrasonic range. To investigate the potential function of ultrasonic call-types in the Humboldt's flying squirrels, we used ultrasonic recorders to record squirrels in the wild across multiple seasons to determine if there are temporal and seasonal patterns in nightly vocal activity and rates of different call-types. We recorded Humboldt's flying squirrels in two geographic locations - Humboldt and San Bernardino counties-in California from 2018 to 2022 in the summer and winter across multiple study areas. We found that although seasonal weather conditions differ between locations, flying squirrels in Humboldt and San Bernardino had similar vocal activity patterns across nightly active periods between locations and between summer and winter. Nightly activity patterns of when the three main chirp-like call-types (arc chirps, tonal chirps, upsweeps) were given varied between seasons in both geographic locations, and these call-types were given at greater rates in the summer in San Bernardino, but rates did not vary by season in Humboldt. Trills, the most structurally complex of the four main call-types, were produced more in the summer than in winter, and also differed in their nightly activity patterns, in both geographic locations. Flying squirrels may use certain call-types earlier or later in the nightly active period due to their potentially varying functions, and may produce more trills in the summer coinciding with the breeding season. Further understanding of the function of different call-types can provide insight into social, foraging, and antipredator behavior of this nocturnal and elusive species.

Context or arousal? Function of drumming in Mongolian gerbils (Meriones unguiculatus).

Drumming is a non-vocal auditory display producing airborne as well as seismic vibrations by tapping body extremities on a surface. It is mostly described as an alarm signal but is also discussed to signal dominance or mating quality. To clarify the function of drumming in Mongolian gerbils (Meriones unguiculatus), we compared the occurrence of drumming during predator, opposite-sex and same-sex encounters. We tested 48 captive Mongolian gerbils (Meriones unguiculatus) in three experiments. In predator experiments, subjects were exposed alone or with their cagemate to aerial and terrestrial predator dummies. In social encounter experiments, familiar and unfamiliar male-female dyads and same-sex dyads were confronted. For the same-sex encounters, a dominance index was calculated for each subject based on the number of won and lost conflicts. Drumming and drumming-call combinations were counted, and a multi-parametric sound analysis was performed. In all experiments drumming and drumming-call combinations occurred. In predator experiments, more subjects drummed when confronted with the predator stimulus than in the habituation phase. In social encounter experiments, more subjects drummed when facing an unfamiliar than a familiar conspecific. In addition, the accompanying call type and body posture of the sender differed between experiments. Thus, we suggest that whereas drumming signals an increased arousal state of the sender, the accompanying call type and the body posture signal context specific information.

Bidirectional generative adversarial representation learning for natural stimulus synthesis.

Thousands of species use vocal signals to communicate with one another. Vocalizations carry rich information, yet characterizing and analyzing these complex, high-dimensional signals is difficult and prone to human bias. Moreover, animal vocalizations are ethologically relevant stimuli whose representation by auditory neurons is an important subject of research in sensory neuroscience. A method that can efficiently generate naturalistic vocalization waveforms would offer an unlimited supply of stimuli with which to probe neuronal computations. Although unsupervised learning methods allow for the projection of vocalizations into low-dimensional latent spaces learned from the waveforms themselves, and generative modeling allows for the synthesis of novel vocalizations for use in downstream tasks, we are not aware of any model that combines these tasks to synthesize naturalistic vocalizations in the waveform domain for stimulus playback. In this paper, we demonstrate BiWaveGAN: a bidirectional generative adversarial network (GAN) capable of learning a latent representation of ultrasonic vocalizations (USVs) from mice. We show that BiWaveGAN can be used to generate, and interpolate between, realistic vocalization waveforms. We then use these synthesized stimuli along with natural USVs to probe the sensory input space of mouse auditory cortical neurons. We show that stimuli generated from our method evoke neuronal responses as effectively as real vocalizations, and produce receptive fields with the same predictive power. BiWaveGAN is not restricted to mouse USVs but can be used to synthesize naturalistic vocalizations of any animal species and interpolate between vocalizations of the same or different species, which could be useful for probing categorical boundaries in representations of ethologically relevant auditory signals.NEW & NOTEWORTHY A new type of artificial neural network is presented that can be used to generate animal vocalization waveforms and interpolate between them to create new vocalizations. We find that our synthetic naturalistic stimuli drive auditory cortical neurons in the mouse equally well and produce receptive field features with the same predictive power as those obtained with natural mouse vocalizations, confirming the quality of the stimuli produced by the neural network.

Agomelatine Is Unable to Attenuate Kainic Acid-Induced Deficits in Early Life Communicative Behavior.

Early life seizures are associated with a variety of behavioral comorbidities. Among the most prevalent of these are deficits in communication. Auditory communicative behaviors in mice, known as ultrasonic vocalizations (USVs), can be used to assess potential treatments. Agomelatine is a melatonin agonist that effectively reduces behavioral comorbidities of seizures in adults; however, its ability to attenuate seizure-induced communicative deficits in neonates is unknown. To address this, we administered C57 mice either saline or kainic acid (KA) on postnatal day (PD) 10. The mice then received either agomelatine or saline 1-h post-status epilepticus. On PD 11, we assessed the quantity of USVs produced, the duration, peak frequency, fundamental frequency, and amplitude of the vocalizations, as well as the call type utilization. We found that KA increased vocal production and reduced USV variability relative to controls. KA also increased USV duration and amplitude and significantly altered the types of calls produced. Agomelatine did not attenuate any of the deficits. Our study is the first to assess agomelatine's efficacy to correct USVs and thus provides an important point of context to the literature, indicating that despite its high therapeutic efficacy to attenuate other behavioral comorbidities of seizures, agomelatine's ability to correct neonatal communicative deficits is limited.

Vocal recognition of partners by female prairie voles.

Recognizing conspecifics is vitally important for differentiating kin, mates, offspring and social threats.1 Although often reliant upon chemical or visual cues, individual recognition across the animal kingdom is also facilitated by unique acoustic signatures in vocalizations.2-4 However, amongst the large Muroidea superfamily of rodents that encompasses laboratory species amenable to neurobiological studies, there is scant behavioral evidence for individual vocal recognition despite individual acoustic variation.5-10 Playback studies have found evidence for coarse communicative functions like mate attraction and territorial defense, but limited finer ability to discriminate known individuals' vocalizations.11-17 Such a capacity would be adaptive for species that form lifelong pair bonds requiring partner identification across timescales, distances and sensory modalities, so to improve the chance of finding individual vocal recognition in a Muroid rodent, we investigated vocal communication in the prairie vole (Microtus ochrogaster) - one of the few socially monogamous mammals.18 We found that the ultrasonic vocalizations of adult prairie voles can communicate individual identity. Even though the vocalizations of individual males change after cohabitating with a female to form a bond, acoustic variation across individuals is greater than within an individual so that vocalizations of different males in a common context are identifiable above chance. Critically, females behaviorally discriminate their partner's vocalizations over a stranger's, even if emitted to another stimulus female. These results establish the acoustic and behavioral foundation for individual vocal recognition in prairie voles, where neurobiological tools19-22 enable future studies revealing its causal neural mechanisms.

A case of chorea-acanthocytosis with significant improvement of symptoms at one year with deep brain stimulation: case report and literature review.

Chorea-acanthocytosis (ChAc) is a rare, neurodegenerative disorder caused by mutations in the VPS13A gene. In this article, we report on a 32-year-old man diagnosed with ChAc, with involuntary movements of the mouth and trunk, drooling of the mouth, slurred speech, and abnormal vocalizations as the main clinical manifestations. Three weeks after implantation of globus pallidus internal (GPi)-deep brain stimulation (DBS), the patient's symptoms improved significantly. For example, articulation is clear, involuntary trunk movements and salivation have largely disappeared, and abnormal vocalizations have been significantly reduced. After 1 year of follow-up, the improvement in involuntary movement symptoms is essentially the same as before. As far as we know, we are the first to report the relief of involuntary vocalizations in a patient with GPi-DBS treatment, and that salivation and involuntary trunk movements have almost disappeared, and all other symptoms are significantly relieved, which is rare in previous cases. All of the above proves that the treatment of our case with DBS was very successful and that longer term follow-up is critical. We also hope that our case will provide new references and therapeutic ideas for the future treatment of patients with ChAc.

Social and emotional alterations in mice lacking the short dystrophin-gene product, Dp71.

BACKGROUND: The Duchenne and Becker muscular dystrophies (DMD, BMD) are neuromuscular disorders commonly associated with diverse cognitive and behavioral comorbidities. Genotype-phenotype studies suggest that severity and risk of central defects in DMD patients increase with cumulative loss of different dystrophins produced in CNS from independent promoters of the DMD gene. Mutations affecting all dystrophins are nevertheless rare and therefore the clinical evidence on the contribution of the shortest Dp71 isoform to cognitive and behavioral dysfunctions is limited. In this study, we evaluated social, emotional and locomotor functions, and fear-related learning in the Dp71-null mouse model specifically lacking this short dystrophin. RESULTS: We demonstrate the presence of abnormal social behavior and ultrasonic vocalization in Dp71-null mice, accompanied by slight changes in exploratory activity and anxiety-related behaviors, in the absence of myopathy and alterations of learning and memory of aversive cue-outcome associations. CONCLUSIONS: These results support the hypothesis that distal DMD gene mutations affecting Dp71 may contribute to the emergence of social and emotional problems that may relate to the autistic traits and executive dysfunctions reported in DMD. The present alterations in Dp71-null mice may possibly add to the subtle social behavior problems previously associated with the loss of the Dp427 dystrophin, in line with the current hypothesis that risk and severity of behavioral problems in patients increase with cumulative loss of several brain dystrophin isoforms.

Adult rat ultrasonic vocalizations and reward: Effects of propranolol and repeated cocaine administration.

BACKGROUND: Mechanisms underlying psychostimulant euphoria remain poorly understood. In adult rats, positive emotional states are associated with alterations in 50-kHz ultrasonic vocalizations (USVs): specifically, "trill" calls are promoted over "flat" calls. Here, we investigated the effects of acute and repeated cocaine administration, and-based on previous findings with amphetamine-their possible dependence on beta-adrenergic receptors. METHODS: Adult male Long-Evans rats received intraperitoneal drug or saline injections before daily USV recording. Fourteen 50-kHz call subtypes were analyzed. In Experiments 1 and 2, cocaine (1-10 mg/kg) and propranolol (10 mg/kg) were tested alone. In Experiment 3, propranolol/cocaine interactions were sought within a conditioned place preference (CPP) procedure. Experiment 4 investigated acute and chronic cocaine effects (Phase 1), and propranolol/cocaine interactions either in an open field (Phase 2) or within a CPP procedure (Phase 3). RESULTS: In drug-naïve animals, cocaine increased the 50-kHz call rate, with sensitization developing rapidly. After more extended exposure, cocaine now also increased the relative prevalence of trill versus flat calls; effects on other subtypes were also revealed. The beta-blocker propranolol prevented neither cocaine CPP nor cocaine effects on USV emission or locomotion but exerted significant USV-related effects when given alone. CPP magnitude and USV-related measures were uncorrelated. CONCLUSIONS: With long-term intraperitoneal administration, cocaine can alter the relative prevalence of several 50-kHz call subtypes; its ability to promote trill versus flat calls, in particular, is consistent with a positive affect interpretation. Cocaine's behavioral effects (i.e., USV-related, locomotor, CPP) appear independent of beta-adrenergic receptor activity.

The ultrasonic vocalization (USV) syllable profile during neonatal opioid withdrawal and a kappa opioid receptor component to increased USV emissions in female mice.

Rationale: Opioid use during pregnancy can lead to negative infant health outcomes, including neonatal opioid withdrawal syndrome (NOWS). NOWS comprises gastrointestinal, autonomic nervous system, and neurological dysfunction that manifest during spontaneous withdrawal. Variability in NOWS severity necessitates a more individualized treatment approach. Ultrasonic vocalizations (USVs) in neonatal mice are emitted in isolation as a stress response and are increased during opioid withdrawal, thus modeling a negative affective state that can be utilized to test new treatments. Objectives: We sought to identify the behavioral and USV profile, brainstem transcriptomic adaptations, and role of kappa opioid receptors in USVs during neonatal opioid withdrawal. Methods: We employed a third trimester-approximate opioid exposure model, where neonatal inbred FVB/NJ pups were injected twice-daily with morphine (10mg/kg, s.c.) or saline (0.9%, 20 ul/g, s.c.) from postnatal day(P) 1 to P14. This protocol induces reduced weight gain, hypothermia, thermal hyperalgesia, and increased USVs during spontaneous morphine withdrawal. Results: On P14, there were increased USV emissions and altered USV syllables during withdrawal, including an increase in Complex 3 syllables in FVB/NJ females (but not males). Brainstem bulk mRNA sequencing revealed an upregulation of the kappa opioid receptor (Oprk1), which contributes to withdrawal-induced dysphoria. The kappa opioid receptor (KOR) antagonist, nor-BNI (30 mg/kg, s.c.), significantly reduced USVs in FVB/NJ females, but not males during spontaneous morphine withdrawal. Furthermore, the KOR agonist, U50,488h (0.625 mg/kg, s.c.), was sufficient to increase USVs on P10 (both sexes) and P14 (females only) in FVB/NJ mice. Conclusions: We identified an elevated USV syllable, Complex 3, and a female-specific recruitment of the dynorphin/KOR system in increased USVs associated with neonatal opioid withdrawal severity.

The link between non-human primate vocalizations and cognition is not constrained by maturation alone: Evidence from healthy preterm infants.

To acquire language, infants must not only identify the signals of their language(s), but also discover how these signals are connected to meaning. By 3 months of age, infants' native language, non-native languages, and vocalizations of non-human primates support infants' formation of object categories-a building block of cognition. But by 6 months, only the native language exerts this cognitive advantage. Prior work with preterm infants indicates that maturation constrains this developing link between the native language and cognition. Here, we assess whether maturation exerts similar constraints on the influence of non-human primate vocalizations on infant categorization. Cross-sectional growth curve analyses of new data from preterm infants and extant data from fullterm infants indicate that developmental tuning of this signal's influence on categorization is best predicted by infants' chronological age, and not gestational status. This evidence, together with prior work, suggests that as infants tune the initially broad set of signals that support early cognition, they are guided by two independent processes: maturation constrains the expression of a link between their native language and cognition, while the influence of non-linguistic signals are guided by other factors, such as postnatal age and experience.

An adverse rearing environment alters maternal responsiveness to infant ultrasonic vocalizations.

Rodent pups use a variety of ultrasonic vocalizations (USVs) to facilitate maternal care. Importantly, infant USV repertoires are dependent on both the age and early life experiences of the pups. We have shown that an adverse rearing environment modeled with the maternal separation (MS) paradigm alters caregiving behavior but little is known about how pup USVs differentially elicit maternal attention. In the present study, maternal approach towards a vocalizing pup over a non-vocalizing pup was tested in a Y-maze apparatus at two developmental time points over the course of MS. At postnatal day (P)10, MS dams engaged in longer interaction times with vocalizing pups compared to non-vocalizing pup, and this effect was strongest in male pups. As expected at P20, dams did not show a preference for either the vocalizing or non-vocalizing pups regardless of rearing environment; however, MS dams spent a greater amount of time in the center of the apparatus as compared to control dams, which can be interpreted as a measure of uncertainty or indecision. These effects of MS on dam USV sensitivity are important considering the sex specific effects of MS exposure across all developmental stages. Our novel findings support the hypothesis that sex-specific pup-dam interactions may drive later life outcomes following adversity.

Ultrasonic vocalization emission is altered following neonatal hypoxic-ischemic brain injury in mice.

Neonatal hypoxic-ischemic (HI) brain injury leads to cognitive impairments including social communication disabilities. Current treatments do not sufficiently target these impairments, therefore new tools are needed to examine social communication in models for neonatal brain injury. Ultrasonic vocalizations (USVs) during early life show potential as a measurement for social development and reflect landmark developmental stages in neonatal mice. However, changes in USV emission early after HI injury have not been found yet. Our current study examines USV patterns and classes in the first 3 days after HI injury. C57Bl/6 mice were subjected to HI on postnatal day (P)9 and USVs were recorded between P10 and P12. Audio files were analyzed using the VocalMat automated tool. HI-injured mice emitted less USVs, for shorter durations, and at a higher frequency compared to control (sham-operated) littermates. The HI-induced alterations in USVs were most distinct at P10 and in the frequency range of 50-75 kHz. At P10 HI-injured mouse pups also produced different ratios of USV class types compared to control littermates. Moreover, alterations in the duration and frequency were specific to certain USV classes in HI animals compared to controls. Injury in the striatum and hippocampus contributed most to alterations in USV communication after HI. Overall, neonatal HI injury leads to USV alterations in newborn mice which could be used as a tool to study early HI-related social communication deficits.