Parental brain through time: The origin and development of the neural circuit of mammalian parenting.

This review consolidates current knowledge on mammalian parental care, focusing on its neural mechanisms, evolutionary origins, and derivatives. Neurobiological studies have identified specific neurons in the medial preoptic area as crucial for parental care. Unexpectedly, these neurons are characterized by the expression of molecules signaling satiety, such as calcitonin receptor and BRS3, and overlap with neurons involved in the reproductive behaviors of males but not females. A synthesis of comparative ecology and paleontology suggests an evolutionary scenario for mammalian parental care, possibly stemming from male-biased guarding of offspring in basal vertebrates. The terrestrial transition of tetrapods led to prolonged egg retention in females and the emergence of amniotes, skewing care toward females. The nocturnal adaptation of Mesozoic mammalian ancestors reinforced maternal care for lactation and thermal regulation via endothermy, potentially introducing metabolic gate control in parenting neurons. The established maternal care may have served as the precursor for paternal and cooperative care in mammals and also fostered the development of group living, which may have further contributed to the emergence of empathy and altruism. These evolution-informed working hypotheses require empirical validation, yet they offer promising avenues to investigate the neural underpinnings of mammalian social behaviors.

Anxious about rejection, avoidant of neglect: Infant marmosets tune their attachment based on individual caregiver's parenting style.

Children's secure attachment with their primary caregivers is crucial for physical, cognitive, and emotional maturation. Yet, the causal links between specific parenting behaviors and infant attachment patterns are not fully understood. Here we report infant attachment in New World monkeys common marmosets, characterized by shared infant care among parents and older siblings and complex vocal communications. By integrating natural variations in parenting styles and subsecond-scale microanalyses of dyadic vocal and physical interactions, we demonstrate that marmoset infants signal their needs through context-dependent call use and selective approaches toward familiar caregivers. The infant attachment behaviors are tuned to each caregiver's parenting style; infants use negative calls when carried by rejecting caregivers and selectively avoid neglectful and rejecting caregivers. Family-deprived infants fail to develop such adaptive uses of attachment behaviors. With these similarities with humans, marmosets offer a promising model for investigating the biological mechanisms of attachment security.

Distinct roles of amylin and oxytocin signaling in intrafamilial social behaviors at the medial preoptic area of common marmosets.

Calcitonin receptor (Calcr) and its brain ligand amylin in the medial preoptic area (MPOA) are found to be critically involved in infant care and social contact behaviors in mice. In primates, however, the evidence is limited to an excitotoxic lesion study of the Calcr-expressing MPOA subregion (cMPOA) in a family-living primate species, the common marmoset. The present study utilized pharmacological manipulations of the cMPOA and shows that reversible inactivation of the cMPOA abolishes infant-care behaviors in sibling marmosets without affecting other social or non-social behaviors. Amylin-expressing neurons in the marmoset MPOA are distributed in the vicinity of oxytocin neurons in the anterior paraventricular nucleus of the hypothalamus. While amylin infusion facilitates infant carrying selectively, an oxytocin's inverse agonist, atosiban, reduces physical contact with non-infant family members without grossly affecting infant care. These data suggest that the amylin and oxytocin signaling mediate intrafamilial social interactions in a complementary manner in marmosets.

Behavioral and histochemical characterization of sexually dimorphic responses to acute social isolation and reunion in mice.

In many mammalian species, females exhibit higher sociability and gregariousness than males, presumably due to the benefit of group living for maternal care. We have previously reported that adult female mice exhibit contact-seeking behaviors upon acute social isolation via amylin-calcitonin receptor (Calcr) signaling in the medial preoptic area (MPOA). In this study, we examined the sex differences in the behavioral responses to acute social isolation and reunion, and the levels of amylin and Calcr expression in the MPOA. We found that male mice exhibited significantly less contact-seeking upon social isolation. Upon reunion, male mice contacted each other to a similar extent as females, but their interactions were more aggressive and less affiliative compared with females. While Calcr-expressing neurons were activated during social contacts in males as in females, the amylin and Calcr expression were significantly lower in males than in females. Together with our previous findings, these findings suggested that the lower expression of both amylin and Calcr may explain the lower contact-seeking and social affiliation of male mice.

[Neurobiology of Loneliness, Social Isolation, and Affiliative Social Behavior].

Prolonged social isolation has been reported to be one of the risk factors for human health, equivalent to smoking cigarettes. Therefore, some developed countries have recognized prolonged social isolation as a social problem and have started to address this problem. Studies on rodent models are essential to fundamentally clarify the impacts of social isolation on human health mentally and physically. In this review, we conduct an overview of the neuromolecular mechanisms of loneliness, perceived social isolation, and the effects of prolonged social isolation. Finally, we consider the evolutionary development of neural bases of loneliness.

Maternal physiological calming responses to infant suckling at the breast.

The mother-infant relation is key to infant physical, cognitive and social development. Mutual regulation and cooperation are required to maintain the dyadic system, but the biological foundation of these responses remains to be clarified. In this study, we report the maternal calming responses to infant suckling during breastfeeding. Using behavioral measures and a Holter electrocardiogram as a readout of the maternal autonomic nervous system, the maternal activities during resting, sitting with her infant on her lap, and breastfeeding were assessed. We found that during breastfeeding, mothers talked less and maternal heart rate was lower than during sitting with the infant without breastfeeding. Congruently, maternal heart rate variability measurements indicated a higher parasympathetic activity during breastfeeding. Time-locked analyses suggested that this maternal calming response was initiated by the tactile stimulation at the breast by the infant face or mouth latch, which preceded the perceived milk ejection. These findings suggest that somatosensory stimuli of breastfeeding activate parasympathetic activity in mothers. Just as how the infant Transport Response facilitates the carrying of infants, the maternal calming responses during breastfeeding may promote efficient milk intake by inhibiting spontaneous maternal activities.

Effects of oxytocin ablation on pup rescue, nursing behaviors and response to pup separation in early-to-mid postpartum mice.

Oxytocin, a neuropeptide hormone, is indispensable for milk ejection during nursing and is important for uterine contractions during parturition. The exact functions of oxytocin in postpartum maternal behaviors and motivations require further investigation. To this end, we characterized the role of oxytocin in components of maternal motivations during the mid-postpartum period, which has not been previously studied. To maintain suckling stimuli, postpartum oxytocin knockout (Oxt-/- ) and heterozygous (Oxt+/- ) littermates were co-housed with a wild-type lactating mother and its litter, and were examined for their ability to retrieve pups under standard or high-risk conditions, nursing behavior, maternal aggression towards an unfamiliar intruder, and motivation to regain contact with separated pups. One-third of Oxt-/- mothers exhibited prolonged parturition but were otherwise grossly healthy. Despite their inability to eject milk, Oxt-/- mothers displayed nursing behaviors for similar durations to Oxt+/- mothers during the second postpartum week. In addition, Oxt-/- mothers were essentially intact for pup retrieval under standard conditions and were motivated to stay close to pups, although they showed a mild decrease in maternal care under high-risk conditions and increased anxiety-like behaviors in pup-related contexts. The present findings indicate that oxytocin is dispensable for nursing behavior and maternal motivations, yet suggest that oxytocin may be relevant for stress resilience in the postpartum period.

Role of Calcr expressing neurons in the medial amygdala in social contact among females.

Social animals become stressed upon social isolation, proactively engaging in affiliative contacts among conspecifics after resocialization. We have previously reported that calcitonin receptor (Calcr) expressing neurons in the central part of the medial preoptic area (cMPOA) mediate contact-seeking behaviors in female mice. Calcr neurons in the posterodorsal part of the medial amygdala (MeApd) are also activated by resocialization, however their role in social affiliation is still unclear. Here we first investigated the functional characteristics of MeApd Calcr + cells; these neurons are GABAergic and show female-biased Calcr expression. Next, using an adeno-associated virus vector expressing a short hairpin RNA targeting Calcr we aimed to identify its molecular role in the MeApd. Inhibiting Calcr expression in the MeApd increased social contacts during resocialization without affecting locomotor activity, suggesting that the endogenous Calcr signaling in the MeApd suppresses social contacts. These results demonstrate the distinct roles of Calcr in the cMPOA and MeApd for regulating social affiliation.

A calcitonin receptor-expressing subregion of the medial preoptic area is involved in alloparental tolerance in common marmosets.

Like humans, common marmoset monkeys utilize family cooperation for infant care, but the neural mechanisms underlying primate parental behaviors remain largely unknown. We investigated infant care behaviors of captive marmosets in family settings and caregiver-infant dyadic situations. Marmoset caregivers exhibited individual variations in parenting styles, comprised of sensitivity and tolerance toward infants, consistently across infants, social contexts and multiple births. Seeking the neural basis of these parenting styles, we demonstrated that the calcitonin receptor-expressing neurons in the marmoset medial preoptic area (MPOA) were transcriptionally activated during infant care, as in laboratory mice. Further, site-specific neurotoxic lesions of this MPOA subregion, termed the cMPOA, significantly reduced alloparental tolerance and total infant carrying, while sparing general health and other social or nonsocial behaviors. These results suggest that the molecularly-defined neural site cMPOA is responsible for mammalian parenting, thus provide an invaluable model to study the neural basis of parenting styles in primates.

A method to soothe and promote sleep in crying infants utilizing the transport response.

Approximately 20%-30% of infants cry excessively and exhibit sleep difficulties for no apparent reason, causing parental stress and even triggering impulsive child maltreatment in a small number of cases.1-8 While several sleep training methods or parental education programs may provide long-term improvement of infant cry and sleep problems, there is yet to be a conclusive recommendation for on-site behavioral interventions.9-13 Previously we have reported that brief carrying of infants transiently reduces infant cry via the transport response, a coordinated set of vagal activation and behavioral calming conserved in altricial mammals.14-18 In this study, we disentangled complex infant responses to maternal holding and transport by combining subsecond-scale, event-locked physiological analyses with dynamic mother-infant interactions. Infant cry was attenuated either by maternal carrying or by reciprocal motion provided by a moving cot, but not by maternal holding. Five-minute carrying promoted sleep for crying infants even in the daytime when these infants were usually awake, but not for non-crying infants. Maternal laydown of sleeping infants into a cot exerted bimodal effects, either interrupting or deepening the infants' sleep. During laydown, sleeping infants were alerted most consistently by the initiation of maternal detachment, then calmed after the completion of maternal detachment in a successful laydown. Finally, the sleep outcome after laydown was associated with the sleep duration before the laydown onset. These data propose a "5-min carrying, 5- to 8- min sitting" scheme for attending to infant cry and sleep difficulties, which should be further substantiated in future studies. VIDEO ABSTRACT.

Amylin-Calcitonin receptor signaling in the medial preoptic area mediates affiliative social behaviors in female mice.

Social animals actively engage in contact with conspecifics and experience stress upon isolation. However, the neural mechanisms coordinating the sensing and seeking of social contacts are unclear. Here we report that amylin-calcitonin receptor (Calcr) signaling in the medial preoptic area (MPOA) mediates affiliative social contacts among adult female mice. Isolation of females from free social interactions first induces active contact-seeking, then depressive-like behavior, concurrent with a loss of Amylin mRNA expression in the MPOA. Reunion with peers induces physical contacts, activates both amylin- and Calcr-expressing neurons, and leads to a recovery of Amylin mRNA expression. Chemogenetic activation of amylin neurons increases and molecular knockdown of either amylin or Calcr attenuates contact-seeking behavior, respectively. Our data provide evidence in support of a previously postulated origin of social affiliation in mammals.

Oxytocin Facilitates Allomaternal Behavior under Stress in Laboratory Mice.

Oxytocin (Oxt) controls reproductive physiology and various kinds of social behaviors, but the exact contribution of Oxt to different components of parental care still needs to be determined. Here, we illustrate the neuroanatomical relations of the parental nurturing-induced neuronal activation with magnocellular Oxt neurons and fibers in the medial preoptic area (MPOA), the brain region critical for parental and alloparental behaviors. We used genetically-targeted mouse lines for Oxt, Oxt receptor (Oxtr), vasopressin receptor 1a (Avpr1a), vasopressin receptor 1b (Avpr1b), and thyrotropin-releasing hormone (Trh) to systematically examine the role of Oxt-related signaling in pup-directed behaviors. The Oxtr-Avpr1a-Avpr1b triple knock-out (TKO), and Oxt-Trh-Avpr1a-Avpr1b quadruple KO (QKO) mice were grossly healthy and fertile, except for their complete deficiency in milk ejection and modest deficiency in parturition secondary to maternal loss of the Oxt or Oxtr gene. In our minimal stress conditions, pup-directed behaviors in TKO and QKO mothers and fathers, virgin females and males were essentially indistinguishable from those of their littermates with other genotypes. However, Oxtr KO virgin females did show decreased pup retrieval in the pup-exposure assay performed right after restraint stress. This stress vulnerability in the Oxtr KO was abolished by the additional Avpr1b KO. The general stress sensitivity, as measured by plasma cortisol elevation after restraint stress or by the behavioral performance in the open field (OF) and elevated plus maze (EPM), were not altered in the Oxtr KO but were reduced in the Avpr1b KO females, indicating that the balance of neurohypophysial hormones affects the outcome of pup-directed behaviors.

Calcitonin receptor signaling in the medial preoptic area enables risk-taking maternal care.

Maternal mammals exhibit heightened motivation to care for offspring, but the underlying neuromolecular mechanisms have yet to be clarified. Here, we report that the calcitonin receptor (Calcr) and its ligand amylin are expressed in distinct neuronal populations in the medial preoptic area (MPOA) and are upregulated in mothers. Calcr+ MPOA neurons activated by parental care project to somatomotor and monoaminergic brainstem nuclei. Retrograde monosynaptic tracing reveals that significant modification of afferents to Calcr+ neurons occurs in mothers. Knockdown of either Calcr or amylin gene expression hampers risk-taking maternal care, and specific silencing of Calcr+ MPOA neurons inhibits nurturing behaviors, while pharmacogenetic activation prevents infanticide in virgin males. These data indicate that Calcr+ MPOA neurons are required for both maternal and allomaternal nurturing behaviors and that upregulation of amylin-Calcr signaling in the MPOA at least partially mediates risk-taking maternal care, possibly via modified connectomics of Calcr+ neurons postpartum.

Evolutionary-adaptive and nonadaptive causes of infant attack/desertion in mammals: Toward a systematic classification of child maltreatment.

Behaviors comparable to human child maltreatment are observed widely among mammals, in which parental care is mandatory for offspring survival. This article first reviews the recent findings on the neurobiological mechanisms for nurturing (infant caregiving) behaviors in mammals. Then the major causes of attack/desertion toward infants (conspecific young) in nonhuman mammals are classified into five categories. Three of the categories are 'adaptive' in terms of reproductive fitness: (i) attack/desertion toward non-offspring; (ii) attack/desertion toward biological offspring with low reproductive value; and (iii) attack/desertion toward biological offspring under unfavorable environments. The other two are nonadaptive failures of nurturing motivation, induced by: (iv) caregivers' inexperience; or (v) dysfunction in caregivers' brain mechanisms required for nurturing behavior. The proposed framework covering both adaptive and nonadaptive factors comprehensively classifies the varieties of mammalian infant maltreatment cases and will support the future development of tailored preventive measures for each human case. Also included are remarks that are relevant to interpretation of available animal data to humans: (1) any kind of child abuse/neglect is not justified in modern human societies, even if it is widely observed and regarded as adaptive in nonhuman animals from the viewpoint of evolutionary biology; (2) group-level characteristics cannot be generalized to individuals; and (3) risk factors are neither deterministic nor irreversible.

Using maternal rescue of pups in a cup to investigate mother-infant interactions in mice/rodents.

Efficient parental care is indispensable for survival of the mammalian offspring, and therefore both parents and offspring cooperate to achieve the best performance. For example, when parents transport altricial offspring, the offspring immediately respond by reducing its cry and movement in both human infants and rodent pups. This coordinated set of central, motor and cardiac responses is designated as the Transport Response (TR) and is shown to facilitate maternal carrying in rodents. The present study aims to investigate the core behavioural characteristics of mother-infant interaction, and to investigate the mechanisms underlying the mother-pup cooperation using pharmacological and genetic manipulations (i.e. Oprm1-/). Along with the clear developmental changes of the pups' immobility and posture during maternal carrying as previously reported, there were also adaptations in maternal strategies, particularly in positioning of foothold and oral grasp over the pup's body, with the pups' age and pup's behaviour. Tree-based models elucidated that both of these maternal variables as well as percentage of pups' struggle predict the time required for pup retrieval from a cup. When the sensory-motor control in pups was disturbed by pharmacological or genetic manipulations, these core behaviours were inefficiently performed and impede maternal retrieval. Mother-infant mutual fit is a complex construct where several intermingled mechanisms are involved. Thus mothers and infants, when interacting, should be considered together as one whole system in which any change in one side or the other, affects the output of the whole dyad. The outcome of the interaction relays on a specific dynamic pattern of infant and maternal behaviours, which mutually change and adapt to fit each other's needs. Key features to reach a successful outcome of the interaction were the maternal retrieving strategy and infants' Transport Response behaviour.

Corticotropin-Releasing Factor Receptor 1 in the Anterior Cingulate Cortex Mediates Maternal Absence-Induced Attenuation of Transport Response in Mouse Pups.

A human infant initially shows non-selective sociality, and gradually develops selective attachment toward its caregiver, manifested as "separation anxiety." It was unclear whether such sophistication of attachment system occurs in non-human mammals. To seek a mouse model of separation anxiety, we utilized a primitive attachment behavior, the Transport Response, in that both human and mouse newborns immediately stop crying and stay immobile to cooperate with maternal carrying. We examined the mouse Transport Response in three social contexts: 30-min isolation in a novel environment, 30-min maternal absence experienced with littermates in the home cage, and the control home-cage condition with the mother and littermates. The pups after postnatal day (PND) 13 attenuated their Transport Response not only in complete isolation but also by maternal absence, and activated several brain areas including the periventricular nucleus of the hypothalamus, suggesting that 30-min maternal absence was perceived as a social stress by mouse pups after PND13. This attenuation of Transport Response by maternal absence was independent with plasma corticosterone, but was diminished by prior administration of a corticotropin-releasing factor receptor 1 (CRFR1) antagonist. Among 18 brain areas examined, only neurons in the anterior cingulate cortex (ACC) co-express c-fos mRNA and CRFR1 after maternal absence. Consistently, excitotoxic ACC lesions inhibited the maternal absence-induced attenuation of Transport Response. These data indicate that the expression of mouse Transport Response is influenced not only by social isolation but also by maternal absence even in their home cage with littermates after PND13, at least partly via CRF-CRFR1 signaling in the ACC.

Oxytocin and Parental Behaviors.

The oxytocin/vasopressin ancestor molecule has been regulating reproductive and social behaviors for more than 500 million years. In all mammals, oxytocin is the hormone indispensable for milk-ejection during nursing (maternal milk provision to offspring), a process that is crucial for successful mammalian parental care. In laboratory mice, a remarkable transcriptional activation occurs during parental behavior within the anterior commissural nucleus (AC), the largest magnocellular oxytocin cell population within the medial preoptic area (although the transcriptional activation was limited to non-oxytocinergic neurons in the AC). Furthermore, there are numerous recent reports on oxytocin's involvement in positive social behaviors in animals and humans. Given all those, the essential involvement of oxytocin in maternal/parental behaviors may seem obvious, but basic researchers are still struggling to pin down the exact role oxytocin plays in the regulation of parental behaviors. A major aim of this review is to more clearly define this role. The best conclusion at this moment is that OT can facilitate the onset of parental behavior, or parental behavior under stressful conditions.In this chapter, we will first review the basics of rodent parental behavior. Next, the neuroanatomy of oxytocin systems with respect to parental behavior in laboratory mice will be introduced. Then, the research history on the functional relationship between oxytocin and parental behavior, along with advancements in various techniques, will be reviewed. Finally, some technical considerations in conducting behavioral experiments on parental behavior in rodents will be addressed, with the aim of shedding light on certain pitfalls that should be avoided, so that the progress of research in this field will be facilitated. In this age of populism, researchers should strive to do even more scholarly works with further attention to methodological details.

Development-dependent behavioral change toward pups and synaptic transmission in the rhomboid nucleus of the bed nucleus of the stria terminalis.

Sexually naïve male C57BL/6 mice aggressively bite unfamiliar pups. This behavior, called infanticide, is considered an adaptive reproductive strategy of males of polygamous species. We recently found that the rhomboid nucleus of the bed nucleus of the stria terminalis (BSTrh) is activated during infanticide and that the bilateral excitotoxic lesions of BSTrh suppress infanticidal behavior. Here we show that 3-week-old male C57BL/6 mice rarely engaged in infanticide and instead, provided parental care toward unfamiliar pups, consistent with observations in rats and other rodent species. This inhibition of infanticide at the periweaning period is functional because the next litter will be born at approximately the time of weaning of the previous litter through maternal postpartum ovulation. However, the mechanism of this age-dependent behavioral change is unknown. Therefore, we performed whole-cell patch clamp recordings of BSTrh and compared evoked neurotransmission in response to the stimulation of the stria terminalis of adult and 3-week-old male mice. Although we were unable to detect a significant difference in the amplitudes of inhibitory neurotransmission, the amplitudes and the paired-pulse ratio of evoked excitatory postsynaptic currents differed between adult and 3-week-old mice. These data suggest that maturation of the synaptic terminal in BSTrh that occurred later than 3 weeks after birth may mediate by the adaptive change from parental to infanticidal behavior in male mice.

Distinct preoptic-BST nuclei dissociate paternal and infanticidal behavior in mice.

Paternal behavior is not innate but arises through social experience. After mating and becoming fathers, male mice change their behavior toward pups from infanticide to paternal care. However, the precise brain areas and circuit mechanisms connecting these social behaviors are largely unknown. Here we demonstrated that the c-Fos expression pattern in the four nuclei of the preoptic-bed nuclei of stria terminalis (BST) region could robustly discriminate five kinds of previous social behavior of male mice (parenting, infanticide, mating, inter-male aggression, solitary control). Specifically, neuronal activation in the central part of the medial preoptic area (cMPOA) and rhomboid nucleus of the BST (BSTrh) retroactively detected paternal and infanticidal motivation with more than 95% accuracy. Moreover, cMPOA lesions switched behavior in fathers from paternal to infanticidal, while BSTrh lesions inhibited infanticide in virgin males. The projections from cMPOA to BSTrh were largely GABAergic. Optogenetic or pharmacogenetic activation of cMPOA attenuated infanticide in virgin males. Taken together, this study identifies the preoptic-BST nuclei underlying social motivations in male mice and reveals unexpected complexity in the circuit connecting these nuclei.

The calming effect of maternal carrying in different mammalian species.

Attachment theory postulates that mothers and their infants possess some basic physiological mechanisms that favor their dyadic interaction and bonding. Many studies have focused on the maternal physiological mechanisms that promote attachment (e.g., mothers' automatic responses to infant faces and/or cries), and relatively less have examined infant physiology. Thus, the physiological mechanisms regulating infant bonding behaviors remain largely undefined. This review elucidates some of the neurobiological mechanisms governing social bonding and cooperation in humans by focusing on maternal carrying and its beneficial effect on mother-infant interaction in mammalian species (e.g., in humans, big cats, and rodents). These studies show that infants have a specific calming response to maternal carrying. A human infant carried by his/her walking mother exhibits a rapid heart rate decrease, and immediately stops voluntary movement and crying compared to when he/she is held in a sitting position. Furthermore, strikingly similar responses were identified in mouse rodents, who exhibit immobility, diminished ultra-sonic vocalizations and heart rate. In general, the studies described in the current review demonstrate the calming effect of maternal carrying to be comprised of a complex set of behavioral and physiological components, each of which has a specific postnatal time window and is orchestrated in a well-matched manner with the maturation of the infants. Such reactions could have been evolutionarily adaptive in mammalian mother-infant interactions. The findings have implications for parenting practices in developmentally normal populations. In addition, we propose that infants' physiological response may be useful in clinical assessments as we discuss possible implications on early screening for child psychopathology (e.g., autism spectrum disorders and perinatal brain disorders).