Oxytocin neurons enable social transmission of maternal behaviour.

Maternal care, including by non-biological parents, is important for offspring survival1-8. Oxytocin1,2,9-15, which is released by the hypothalamic paraventricular nucleus (PVN), is a critical maternal hormone. In mice, oxytocin enables neuroplasticity in the auditory cortex for maternal recognition of pup distress15. However, it is unclear how initial parental experience promotes hypothalamic signalling and cortical plasticity for reliable maternal care. Here we continuously monitored the behaviour of female virgin mice co-housed with an experienced mother and litter. This documentary approach was synchronized with neural recordings from the virgin PVN, including oxytocin neurons. These cells were activated as virgins were enlisted in maternal care by experienced mothers, who shepherded virgins into the nest and demonstrated pup retrieval. Virgins visually observed maternal retrieval, which activated PVN oxytocin neurons and promoted alloparenting. Thus rodents can acquire maternal behaviour by social transmission, providing a mechanism for adapting the brains of adult caregivers to infant needs via endogenous oxytocin.

Highly demarcated structural alterations in the brain and impaired social incentive learning in Tbx1 heterozygous mice.

Copy number variants (CNVs) are robustly associated with psychiatric disorders and changes in brain structures. However, because CNVs contain many genes, the precise gene-phenotype relationship remains unclear. Although various volumetric alterations in the brains of 22q11.2 CNV carriers have been identified in humans and mouse models, it is unknown how each gene encoded in the 22q11.2 region contributes to structural alterations, associated mental illnesses, and their dimensions. Our previous studies identified Tbx1, a T-box family transcription factor encoded in the 22q11.2 CNV, as a driver gene for social interaction and communication, spatial and working memory, and cognitive flexibility. However, it remains unclear how TBX1 impacts the volumes of various brain regions and their functionally linked behavioral dimensions. In this study, we used volumetric magnetic resonance imaging analysis to comprehensively evaluate brain region volumes and behavioral alterations relevant to affected structures in congenic Tbx1 heterozygous mice. Our data showed that the volumes of the anterior and posterior portions of the amygdaloid complex and its surrounding cortical regions were most robustly reduced in Tbx1 heterozygous mice. In an amygdala-dependent task, Tbx1 heterozygous mice were impaired in their ability to learn the incentive value of a social partner. The volumes of the primary and secondary auditory cortexes were increased, and acoustic, but not non-acoustic, sensorimotor gating was impaired in Tbx1 heterozygous mice. Our findings identify the brain's regional volume alterations and their relevant behavioral dimensions associated with Tbx1 heterozygosity.

Development of the Paternal Brain in Humans throughout Pregnancy.

Previous studies have demonstrated that paternal caregiving behaviors are reliant on neural pathways similar to those supporting maternal care. Interestingly, a greater variability exists in parental phenotypes in men than in women among individuals and mammalian species. However, less is known about when or how such variability emerges in men. We investigated the longitudinal changes in the neural, hormonal, and psychological bases of expression of paternal caregiving in humans throughout pregnancy and the first 4 months of the postnatal period. We measured oxytocin and testosterone, paternity-related psychological traits, and neural response to infant-interaction videos using fMRI in first-time fathers and childless men at three time points (early to mid-pregnancy, late pregnancy, and postnatal). We found that paternal-specific brain activity in prefrontal areas distinctly develops during middle-to-late pregnancy and is enhanced in the postnatal period. In addition, among fathers, the timing of the development of prefrontal brain activity was associated with specific parenting phenotypes.

Development of the paternal brain in expectant fathers during early pregnancy.

The human parenting brain network mediates caregiving behaviors. When exposed to the stimuli of their infants, compared with non-parents, both fathers and mothers exhibit distinct patterns of neural activation. As human males, relative to females, do not undergo robust physiological changes during pregnancy, when and how the paternal brain networks begin to form remains unclear. Thus, using functional MRI, we examined brain activation in response to infant-interaction videos in two groups, childless males and first-time expectant fathers during their partners' early pregnancy before remarkable changes in their partners' appearances commenced. Multivoxel pattern analysis revealed that expectant fathers' left anterior insula and inferior frontal gyrus showed incipient changes in response to parenthood during early pregnancy. Furthermore, these changes were associated with several paternal traits, such as a negative image toward parenting. Such external factors might influence the paternal brain's development during early pregnancy.

Basal cortisol concentrations related to maternal behavior during puppy development predict post-growth resilience in dogs.

A prolonged stress hyporesponsive period (SHRP) due to the mother's presence may delay the timing of glucocorticoid (GC) elevation in infants, thereby reducing the excessive stress response that would affect post-growth temperament. In dogs, the SHRP has been suggested to persist until postnatal week (PW) 4; therefore, PW 5, which SHRP may be prolonged by the mother dog, may be a critical point in the developmental stage of dogs to establish stress responsiveness. We conducted a long-term survey on the development of dogs to investigate i) whether the degree of the stress response at PW 5 is determined by maternal behavior and ii) whether it can predict post-growth stress responses and temperament in dogs. As a result, the offspring of mother dogs who had more delivery experience and exhibited more maternal behavior showed higher basal cortisol concentrations at PW 5. These offspring may have acquired less fear response as an individual trait and had relatively quick adaptability, albeit with high cortisol concentrations during exposure to novel environments post-growth, suggesting that high cortisol concentrations at PW 5 are linked to resilience post-growth. Basal cortisol concentrations at PW 7 were not affected by maternal variables and were not associated with cortisol response to novel environments post-growth. GCs are essential hormones that increase the probability of survival. Therefore, the high hypothalamic pituitary adrenal activities of the mother dogs and their offspring in this study may not immediately indicate negative states, and these results prompt a reconsideration of the role of GC in organisms.

Responses to pup vocalizations in subordinate naked mole-rats are induced by estradiol ingested through coprophagy of queen's feces.

Naked mole-rats form eusocial colonies consisting of a single breeding female (the queen), several breeding males, and sexually immature adults (subordinates). Subordinates are cooperative and provide alloparental care by huddling and retrieving pups to the nest. However, the physiological mechanism(s) underlying alloparental behavior of nonbreeders remains undetermined. Here, we examined the response of subordinates to pup voice and the fecal estradiol concentrations of subordinates during the three reproductive periods of the queen, including gestation, postpartum, and nonlactating. Subordinate response to pup voice was observed only during the queen's postpartum and was preceded by an incremental rise in subordinates' fecal estradiol concentrations during the queen's gestation period, which coincided with physiological changes in the queen. We hypothesized that the increased estradiol in the queen's feces was disseminated to subordinates through coprophagy, which stimulated subordinates' responses to pup vocalizations. To test this hypothesis, we fed subordinates either fecal pellets from pregnant queens or pellets from nonpregnant queens amended with estradiol for 9 days and examined their response to recorded pup voice. In both treatments, the subordinates exhibited a constant level of response to pup voice during the feeding period but became more responsive 4 days after the feeding period. Thus, we believe that we have identified a previously unknown system of communication in naked mole-rats, in which a hormone released by one individual controls the behavior of another individual and influences the level of responsiveness among subordinate adults to pup vocal signals, thereby contributing to the alloparental pup care by subordinates.

Testosterone regulates the emission of ultrasonic vocalizations and mounting behavior during different developmental periods in mice.

Testosterone masculinizes male sexual behavior by providing organizational and activational effects during the perinatal and peripubertal periods and during adulthood, respectively. We revealed that the emission of ultrasonic vocalizations (USVs) and mounting behavior was regulated by different neural circuits. However, the detailed testosterone effects on these two behaviors have not been fully elucidated. Here, we evaluated the time-dependent effects of testosterone on USVs and mounting behavior in mice using a testosterone treatment model, in which females were treated with testosterone to assess the "gain-of-function" and a "loss-of-function" model. In the loss-of-function model, we used Ad4BP/SF-1ΔFLC/- male mice, in which testosterone production was abolished in prenatal and postnatal stages, and Ad4BP/SF-1ΔFLC/ΔFLC mice, in which testosterone production was markedly reduced only in prenatal stages. When testosterone was administered to female mice during the neonatal and peripubertal periods, but not during adulthood, USV emissions increased. Conversely, testosterone treatment in adult female mice increased the mounting behavior, but not USVs. In Ad4BP/SF-1ΔFLC/- mice, USVs and mounting behavior was completely absent. Ad4BP/SF-1ΔFLC/ΔFLC male mice displayed equivalent levels of USVs but less mounting behavior. Collectively, these results suggest that testosterone has dual regulatory roles in USV emissions and mounting behavior.

Maternal approach behaviors toward neonatal calls are impaired by mother's experiences of raising pups with a risk gene variant for autism.

How the intrinsic sequence structure of neonatal mouse pup ultrasonic vocalization (USV) and maternal experiences determine maternal behaviors in mice is poorly understood. Our previous work showed that pups with a Tbx1 heterozygous (HT) mutation, a genetic risk for autism spectrum disorder (ASD), emit altered call sequences that do not induce maternal approach behaviors in C57BL6/J mothers. Here, we tested how maternal approach behaviors induced by wild-type and HT USVs are influenced by the mother's experience in raising pups of these two genotypes. The results showed that wild-type USVs were effective in inducing maternal approach behaviors when mothers raised wild-type but not HT pups. The USVs of HT pups were ineffective regardless of whether mothers raised HT or wild-type pups. However, the sequence structure of pup USVs had no effect on the general, non-directional incentive motivation of maternal behaviors. Our data show how the mother's experience with a pup with a genetic risk for ASD alters the intrinsic incentive values of USV sequences in maternal approach behaviors.

Breastfeeding dynamically changes endogenous oxytocin levels and emotion recognition in mothers.

Breastfeeding behaviours can significantly change mothers' physiological and psychological states. The hormone oxytocin may mediate breastfeeding and mothers' emotion recognition. This study examined the effects of endogenous oxytocin fluctuation via breastfeeding on emotion recognition in 51 primiparous mothers. Saliva oxytocin was assessed before and after the manipulation (breastfeeding or holding an infant), and emotion recognition tasks were conducted. Among mothers who breastfed daily, mothers with more increased levels of oxytocin after breastfeeding showed more reduced negative recognition and enhanced positive recognition of adult facial expressions. These oxytocin functions accompanying breastfeeding may support continued nurturing behaviours and also affect the general social cognition of other adults beyond any specific effect on infants.

Gonadal steroid hormone secretion during the juvenile period depends on host-specific microbiota and contributes to the development of odor preference.

The host microbial community is thought to have an important role in the host endocrine system and behavioral phenotype. We investigated chronological changes of levels of gonadal hormones and corticosterone in the feces of 4- to 8-week-old female germ-free (GF) mice, and conducted odor preference test at 8 weeks of age. We further evaluated the developmental impact of the microbial community by analyzing 4-week-old GF mice orally administered the fecal microbiota of specific pathogen-free (SPF) mice or guinea pigs (GF-SPF mice or GF-Guinea pig mice). The fecal estradiol, progesterone, and corticosterone levels of GF mice were lower than those of SPF mice. Furthermore, the increased levels in GF mice were suggested to be caused by colonization of microbiota of SPF mice or guinea pigs. However, the degree of recovery of progesterone and corticosterone by microbiota of guinea pigs was lower than that by SPF mice. In odor preference tests, interestingly, female GF mice preferred female odors to male odors, although this preference was not seen in other mice. These findings suggested that the microbial community plays an important role in the development of the host endocrine system for gonadal hormones and corticosterone, and odor preference in mice.

Effect of Canine Oxytocin Receptor Gene Polymorphism on the Successful Training of Drug Detection Dogs.

Drug detection dogs can be trained to locate various prohibited drugs with targeted odors, and they play an important role in the interdiction of drug smuggling in human society. Recent studies provide the interesting hypothesis that the oxytocin system serves as a biological basis for co-evolution between dogs and humans. Here, we offer the new possibility that genetic variation of the canine oxytocin receptor (OXTR) gene may regulate the success of a dog's training to become a drug detection dog. A total of 340 Labrador Retriever dogs that were trained to be drug detection dogs in Japan were analyzed. We genotyped an exonic SNP (rs8679682) in the OXTR gene and compared the training success rate of dogs with different genotypes. We also asked dog trainers in the training facility to evaluate subjective personality assessment scores for each dog and examined how each dog's training success was related to those scores. A significant effect of the OXTR genotype on the success of the dogs' training was found, with a higher proportion of dogs carrying the C allele (T/C and C/C genotypes) being successful candidates than dogs carrying the T/T genotype. Dog personality scores of Training Focus (Factor 1) were positively correlated with an increased likelihood that a dog would successfully complete training. Although the molecular mechanism of the OXTR gene and its functional pathway related to dog behavior remains unknown, our findings suggest that canine OXTR gene variants may regulate individual differences between dogs in their responsiveness to training for drug detection.

Male mice ultrasonic vocalizations enhance female sexual approach and hypothalamic kisspeptin neuron activity.

Vocal communication in animals is important for ensuring reproductive success. Male mice emit song-like "ultrasonic vocalizations (USVs)" when they encounter female mice, and females show approach to the USVs. However, it is unclear whether USVs of male mice trigger female behavioral and endocrine responses in reproduction. In this study, we first investigated the relationship between the number of deliveries in breeding pairs for 4months and USVs syllables emitted from those paired males during 3min of sexual encounter with unfamiliar female mice. There was a positive correlation between these two indices, which suggests that breeding pairs in which males could emit USVs more frequently had more offspring. Further, we examined the effect of USVs of male mice on female sexual behavior. Female mice showed more approach behavior towards vocalizing males than devocalized males. Finally, to determine whether USVs of male mice could activate the neural system governing reproductive function in female mice, the activation of kisspeptin neurons, key neurons to drive gonadotropin-releasing hormone neurons in the hypothalamus, was examined using dual-label immunocytochemistry with cAMP response element-binding protein phosphorylation (pCREB). In the arcuate nucleus (Arc), the number of kisspeptin neurons expressing pCREB significantly increased after exposure to USVs of male as compared with noise exposure group. In conclusion, our results suggest that USVs of male mice promote fertility in female mice by activating both their approaching behavior and central kisspeptin neurons.

Oxytocin promotes social bonding in dogs.

Recent evidence suggests that enduring social bonds have fitness benefits. However, very little is known about the neural circuitry and neurochemistry underlying the formation and maintenance of stable social bonds outside reproductive contexts. Oxytocin (OT), a neuropeptide synthetized by the hypothalamus in mammals, regulates many complex forms of social behavior and cognition in both human and nonhuman animals. Animal research, however, has concentrated on monogamous mammals, and it remains unknown whether OT also modulates social bonds in nonreproductive contexts. In this study we provide behavioral evidence that exogenous OT promotes positive social behaviors in the domestic dog toward not only conspecifics but also human partners. Specifically, when sprayed with OT, dogs showed higher social orientation and affiliation toward their owners and higher affiliation and approach behaviors toward dog partners than when sprayed with placebo. Additionally, the exchange of socio-positive behaviors with dog partners triggered the release of endogenous OT, highlighting the involvement of OT in the development of social relationships in the domestic dog. These data provide new insight into the mechanisms that facilitate the maintenance of close social bonds beyond immediate reproductive interest or genetic ties and complement a growing body of evidence that identifies OT as one of the neurochemical foundations of sociality in mammalian species.

Early weaning impairs fear extinction and decreases brain-derived neurotrophic factor expression in the prefrontal cortex of adult male C57BL/6 mice.

We examined the developmental effects of early weaning on anxiety and the extinction of fear memory in male C57BL/6 mice. Early weaning led to increased freezing behaviors after fear conditioning via the foot-shock method both during extinction training and in a test of extinction recall, but did not induce significant changes in anxiety-like behavior. In addition, we found that the levels of brain-derived neurotrophic factor (BDNF) protein and mRNA transcripts for BDNF exon III in the medial prefrontal cortex (mPFC) at the time of extinction recall were impaired in early-weaned mice compared to normally weaned mice. In light of consistent finding that early-weaned mice experience greater stress at weaning, these data suggest that early weaning of male C57BL/6 mice is valuable for studies of the pathogenesis of post-traumatic stress disorder.

Early weaning impairs a social contagion of pain-related stretching behavior in mice.

Pain sensitivity in mice can be modulated through exposure to familiar individuals. This phenomenon is considered a form of emotional contagion, thought to be an evolutionary precursor of empathy in mammals. In particular, mother-infant interactions early in life can considerably alter empathy development. Here, we demonstrated that pairs of mice that were simultaneously administered with a noxious stimulus (acetic acid) exhibited more pain-related behaviors than when one of the pair was treated with a noxious stimulus. However, these differences disappeared when mice were separated from the dam 1 week earlier than the typical weaning age. Even when mice were alone, when treated with acetic acid, early weaning decreased their pain response. These results suggested that the disruption of mother-infant bonding through early weaning impairs pain contagion and modulates sensitivity to pain.

Sex-reversed correlation between stress levels and dominance rank in a captive non-breeder flock of crows.

Group living has both benefits and costs to individuals; benefits include efficient acquisition of resources, and costs include stress from social conflicts among group members. Such social challenges result in hierarchical dominance ranking among group members as a solution to avoid escalating conflict that causes different levels of basal stress between individuals at different ranks. Stress-associated glucocorticoid (corticosterone in rodents and birds; CORT) levels are known to correlate with dominance rank in diverse taxa and to covary with various social factors, such as sex and dominance maintenance styles. Although there is much evidence for sex differences in the basal levels of CORT in various species, the correlation of sex differences in basal CORT with dominance rank is poorly understood. We investigated the correlation between CORT metabolites (CM) in the droppings and social factors, including rank and sex, in a captive non-breeder group of crows. In this group, all the single males dominated all the single females, and dominance ranks were stable among single males but relatively unstable among single females. CM levels and rank were significantly correlated in a sex-reversed fashion: males at higher rank (i.e., more dominant) had higher CM, whereas females at higher rank exhibited lower CM. This is the first evidence of sex-reversed patterns of CM-rank correlation in birds. The results suggest that different mechanisms of stress-dominance relationships operate on the sexes in non-breeder crow aggregations; in males, stress is associated with the cost of aggressive displays, whereas females experience subordination stress due to males' overt aggression.

The behavioral and endocrinological development of stress response in dogs.

Endocrinological stress response has been shown to be absent in a specific period of the early life of rodents; this is named the stress-hyporesponsive period (SHRP). The SHRP is a significant period for the appropriate development of infants. In this study, the presence of SHRP in dogs was identified by conducting a 5-min separation test in 142 Labrador retriever puppies in their early socialization period and measuring the changes in urinary cortisol levels. An increase in cortisol after separation was found after 5 weeks of age, suggesting that the SHRP persists until 4 weeks of age in dogs. The distress vocalization during separation changed and the lactating behavior decreased rapidly around 5 weeks of age, suggesting that the endocrinological and emotional aspects of development change at approximately 5 weeks of age and maternal inhibition of cortisol might occur in dogs as well as rodents.

Oxytocin receptor control social information about fear expression of others in mice.

The social functions of oxytocin are diverse, and the specific aspects of information processing involved in emotional contagion remain unclear. We compared some fear-related behaviors among oxytocin receptor knockout mice and oxytocin-receptor-reduced mice with that of wild-type mice. In the observational fear assay, which reflects fear emotional contagion, mice that observed other individuals receiving electric shocks exhibited vicarious freezing. Mice with reduced or knockout oxytocin receptor expression showed reduced vicarious freezing. In the emotional discrimination assay, which reflects the ability to perceive others' emotional cues, we compared approach and scent-sniffing behaviors toward fear and emotionally neutral individuals. While wild-type mice were able to detect the fear emotion of others, mice with reduced or knocked-out oxytocin receptors showed reduced discrimination ability. In the fear behavior assays, which do not present social cues, we did not find these differences in oxytocin receptor expression in the brain. These findings indicate that oxytocin plays a role in emotional contagion by perceiving the emotions of others.

Long-term monitoring of huddling behavior in mice using online image processing.

Many animal species, including mice, form societies of numerous individuals for survival. Understanding the interactions between individual animals is crucial for elucidating group behavior. One such behavior in mice is huddling, yet its analysis has been limited. In this study, we propose a cost-effective method for monitoring long-term huddling behavior in mice using online image processing with OpenCV. This method treats a single mouse or a group of mice as a cluster of pixels (a 'blob') in video images, extracting and saving only essential information such as areas, coordinates, and orientations. This approach reduces data storage needs to 1/200000th of what would be required if the video were recorded in its compressed form, thereby enabling long-term behavioral analysis. To validate the performance of our algorithm, ~2000 video frames were randomly chosen. We manually counted the number of clusters of mice in these frames and compared them with the number of blobs automatically detected by the algorithm. The results indicated a high level of consistency, exceeding 90% across the selected video frames. Initial observations of both male and female groups suggested some variations in huddling behavior among male and female groups; however, these results should be interpreted cautiously due to a small sample. Group behavior is known to be disrupted in several neuropsychiatric disorders, such as autism. Various mouse models of these disorders have been proposed. Our measurement system, when combined with drug or genetic modification screening, could provide a valuable tool for high-throughput analyses of huddling behavior.

Analysis of Electrocardiograms and Behavior in Mice from Pregnancy to Lactation Period.

Changes in the mother-offspring relationship are presumably accompanied by dynamic changes in the autonomic nervous system. Although temporal measurements of autonomic activity have been performed in human mothers and infants, the analysis of long-term changes remains unexplored. Mouse mothers can form social bonds with their pups and have a short period of pregnancy and lactation, which makes them useful for the examination of physiological changes from pregnancy to pup-rearing. Therefore, a telemetry system was used for several weeks to measure the changes in the autonomic nervous system and the behavior of mouse mothers. The current results showed that an electrocardiogram (ECG) could be stably recorded regardless of the movements of mothers and parturition. ECG analysis showed that the heart rate gradually decreased from pregnancy to lactation, and sympathetic activity sharply increased as the pups developed. Furthermore, the simultaneous recording of behavior and ECG in the home cage enabled us to understand the behavior-dependent influences on the ECG, thereby revealing the characteristics of autonomic nervous activity during each behavior. Thus, the present experimental method helps to understand how the physiological characteristics of mothers change from pregnancy through pup rearing, supporting the healthy development of pups.