Characterization of oxytocin and vasopressin receptors in the Southern giant pouched rat and comparison to other rodents.

Vasopressin and oxytocin are well known and evolutionarily ancient modulators of social behavior. The distribution and relative densities of vasopressin and oxytocin receptors are known to modulate the sensitivity to these signaling molecules. Comparative work is needed to determine which neural networks have been conserved and modified over evolutionary time, and which social behaviors are commonly modulated by nonapeptide signaling. To this end, we used receptor autoradiography to determine the distribution of vasopressin 1a and oxytocin receptors in the Southern giant pouched rat (Cricetomys ansorgei) brain, and to assess the relative densities of these receptors in specific brain regions. We then compared the relative receptor pattern to 23 other species of rodents using a multivariate ANOVA. Pouched rat receptor patterns were strikingly similar to hamsters and voles overall, despite the variation in social organization among species. Uniquely, the pouched rat had dense vasopressin 1a receptor binding in the caudate-putamen (i.e., striatum), an area that might impact affiliative behavior in this species. In contrast, the pouched rat had relatively little oxytocin receptor binding in much of the anterior forebrain. Notably, however, oxytocin receptor binding demonstrated extremely dense binding in the bed nucleus of the stria terminalis, which is associated with the modulation of several social behaviors and a central hub of the social decision-making network. Examination of the nonapeptide system has the potential to reveal insights into species-specific behaviors and general themes in the modulation of social behavior.

Extreme plasticity of reproductive state in a female rodent.

Successful sexual reproduction relies on the coordination of multiple biological systems, yet traditional concepts of biological sex often ignore the natural plasticity in morphology and physiology underlying sex. Most female mammals develop a patent (i.e., opened) vaginal entrance (introitus) prenatally or postnatally before or during puberty, usually under the influence of estrogens, and remain patent for the remainder of their lifespan1. An exception is the southern African giant pouched rat (Cricetomys ansorgei), whose vaginal introitus remains sealed well into adulthood2. Here, we explore this phenomenon and report that the reproductive organs and the vaginal introitus can undergo astounding and reversible transformation. Non-patency is characterized by reduced uterine size and the presence of a sealed vaginal introitus. Furthermore, the female urine metabolome shows that patent and non-patent females profoundly differ in their urine content, a reflection of differences in physiology and metabolism. Surprisingly, patency state did not predict fecal estradiol or progesterone metabolite concentrations. Exploring the plasticity that exists in reproductive anatomy and physiology can uncover that traits long considered 'fixed' in adulthood can become plastic under specific evolutionary pressures. Moreover, the barriers to reproduction that such plasticity creates present unique challenges to maximizing reproductive potential.

Effects of a GnRH agonist on sex behavior in females of the southern giant pouched rat.

In general, males should be particularly attentive to cues of sexual availability and females should advertise accordingly. Vaginal patency (i.e., the openness of the vagina) is a reliable indicator of sexual maturity; if the vagina is closed, the female is unable to copulate. The southern giant pouched rat (Cricetomys ansorgei) is unusual because females can have fully fused vaginal openings (i.e., vaginal nonpatency) despite being considered 'adults' by other metrics. Moreover, some females reversibly close their vaginal openings. Thus, vaginal patency in the pouched rat is a 'flexible' reproductive state. We subcutaneously implanted a long-acting GnRH agonist (deslorelin), which over time inhibits sex steroid secretion, to better understand the endocrinology and social behavior relating to vaginal patency in this species. We hypothesized that altering GnRH would impact both patency and behavior through its effects on circulating levels of estradiol. Six months of deslorelin treatment did not alter vaginal patency. Behaviorally, deslorelin-treated females spent less time interacting with, and were more aggressive towards males (compared to controls). Notably, deslorelin did not alter female scent marking. We conclude that behavioral receptivity, but not vaginal patency, is impacted by GnRH hormonal cascades in the pouched rat.

The Philosophy of Outliers: Reintegrating Rare Events Into Biological Science.

Individual variation in morphology, physiology, and behavior has been a topic of great interest in the biological sciences. While scientists realize the importance of understanding diversity in individual phenotypes, historically the "minority" results (i.e., outlier observations or rare events) of any given experiment have been dismissed from further analysis. We need to reframe how we view "outliers" to improve our understanding of biology. These rare events are often treated as problematic or spurious, when they can be real rare events or individuals driving evolution in a population. It is our perspective that to understand what outliers can tell us in our data, we need to: (1) Change how we think about our data philosophically, (2) Fund novel collaborations using science "weavers" in our national funding agencies, and (3) Bridge long-term field and lab studies to reveal these outliers in action. By doing so, we will improve our understanding of variation and evolution. We propose that this shift in culture towards more integrative science will incorporate diverse teams, citizen scientists and local naturalists, and change how we teach future students.

Female-Female Reproductive Suppression: Impacts on Signals and Behavior.

Female-female reproductive suppression is evident in an array of mammals, including rodents, primates, and carnivores. By suppressing others, breeding females can benefit by reducing competition from other females and their offspring. There are neuroendocrinological changes during suppression which result in altered behavior, reproductive cycling, and communication. This review, which focuses on species in Rodentia, explores the current theoretical frameworks of female-female reproductive suppression, how female presence and rank impacts reproductive suppression, and some of the proposed mechanisms of suppression. Finally, the understudied role of olfactory communication in female-female reproductive suppression is discussed to identify current gaps in our understanding of this topic.

The giant pouched rat (Cricetomys ansorgei) olfactory receptor repertoire.

For rodents, olfaction is essential for locating food, recognizing mates and competitors, avoiding predators, and navigating their environment. It is thought that rodents may have expanded olfactory receptor repertoires in order to specialize in olfactory behavior. Despite being the largest clade of mammals and depending on olfaction relatively little work has documented olfactory repertoires outside of conventional laboratory species. Here we report the olfactory receptor repertoire of the African giant pouched rat (Cricetomys ansorgei), a Muroid rodent distantly related to mice and rats. The African giant pouched rat is notable for its large cortex and olfactory bulbs relative to its body size compared to other sympatric rodents, which suggests anatomical elaboration of olfactory capabilities. We hypothesized that in addition to anatomical elaboration for olfaction, these pouched rats might also have an expanded olfactory receptor repertoire to enable their olfactory behavior. We examined the composition of the olfactory receptor repertoire to better understand how their sensory capabilities have evolved. We identified 1145 intact olfactory genes, and 260 additional pseudogenes within 301 subfamilies from the African giant pouched rat genome. This repertoire is similar to mice and rats in terms of size, pseudogene percentage and number of subfamilies. Analyses of olfactory receptor gene trees revealed that the pouched rat has 6 expansions in different subfamilies compared to mice, rats and squirrels. We identified 81 orthologous genes conserved among 4 rodent species and an additional 147 conserved genes within the Muroid rodents. The orthologous genes shared within Muroidea suggests that there may be a conserved Muroid-specific olfactory receptor repertoire. We also note that the description of this repertoire can serve as a complement to other studies of rodent olfaction, as the pouched rat is an outgroup within Muroidea. Thus, our data suggest that African giant pouched rats are capable of both natural and trained olfactory behaviors with a typical Muriod olfactory receptor repertoire.

Call-specific patterns of neural activation in auditory processing of Richardson's ground squirrel alarm calls.

INTRODUCTION: Richardson's ground squirrels use alarm calls to warn conspecifics about potential predatory threats. Chirp calls typically indicate high levels of threat from airborne predators, while whistle calls are associated with lower levels of threat from terrestrial predators. These types of calls primarily elicit escape behaviors and increased vigilance in receivers, respectively. While much is known about the neural mechanisms involved in the production of vocalizations, less is known about the mechanisms important for the perception of alarm calls by receivers, and whether changes in perceived risk are associated with unique patterns of neuronal activation. Thus, to determine whether alarm calls associated with different levels of predation risk result in differential neuronal activation, we used immunohistochemistry to identify and quantify c-Fos immunopositive cells in brain regions important in stress, fear, danger, and reward, following alarm call reception. METHODS: We exposed 29 female Richardson's ground squirrels (10 control, 10 whistle receivers, and 9 chirp receivers) to playbacks of whistles, chirps, or a no-vocalization control. We then assessed neuronal activation via c-Fos immunohistochemistry in 12 brain regions. RESULTS: Ground squirrels receiving high-threat "chirp" vocalizations had reduced neuronal activation in the medial amygdala and superior colliculus compared with controls. It is likely that changes in activity in these brain regions serve to alter the balance between approach and avoidance in turn promoting escape behaviors. CONCLUSIONS: Thus, we conclude that in Richardson's ground squirrels, these brain regions are important for the perception of risk resulting from receiving alarm calls and allow for appropriate behavioral responses by receivers.

Comparison of the distribution of oxytocin and vasopressin 1a receptors in rodents reveals conserved and derived patterns of nonapeptide evolution.

Oxytocin (OT) and vasopressin (VP) are known modulators of social behaviour across rodents. Research has revealed the location of action of these nonapeptides through localization of their associated receptors, which include the oxytocin receptor (OTR) and the vasopressin 1a receptor (V1aR). As research into these complex systems has progressed, studies investigating how these systems modulate behaviour have remained relatively narrow in scope (ie, focused on how a single brain region shapes behaviour in only a handful of species). However, the brain regions that regulate social behaviour are part of interconnected neural networks for which coordinated activity enables behavioural variation. Thus, to better understand how nonapeptide systems have evolved under different selective pressures among rodent species, we conducted a meta-analysis using a multivariate comparative method to examine the patterns of OTR and V1aR density expression in this taxon. Several brain regions were highly correlated based on their OTR and V1aR binding patterns across species, supporting the notion that the distribution of these receptors is highly conserved in rodents. However, our results also revealed that specific patterns of V1aR density differed from OTR density, and within-genus variance for V1aR was low compared to between-genus variance, suggesting that these systems have responded and evolved quite differently to selective pressures over evolutionary time. We propose that, in addition to examining single brain regions of interest, taking a broad comparative approach when studying the OT and VP systems is important for understanding how the systemic action of nonapeptides modulate social behaviour across species.

Gone girl: Richardson's ground squirrel offspring and neighbours are resilient to female removal.

Within matrilineal societies, the presence of mothers and female kin can greatly enhance survival and reproductive success owing to kin-biased alarm calling, cooperation in territory defence, protection from infanticidal conspecifics, joint care of young and enhanced access to resources. The removal of mothers by predators or disease is expected to increase the stress experienced by offspring via activation of their hypothalamic-pituitary-adrenal axis, increasing circulating glucocorticoids and reducing offspring survival and reproductive success. Yet, few studies have removed mothers in the post-weaning period to examine the assumed physiological and fitness consequences associated with these mortality events. We examined how the loss of a mother affects juvenile Richardson's ground squirrels' (Urocitellus richardsonii) faecal glucocorticoid metabolites and their survival. Given that neighbours are often close kin, we further hypothesized that conspecific removal would similarly diminish the fitness of neighbouring individuals. Upon removing the mother, we detected no impact on offspring or neighbouring conspecific faecal glucocorticoid metabolites in the removal year, or on overwinter survival in the following year. Furthermore, no impact on neighbour reproductive success was detected. Given the high predation rates of ground squirrels in wild populations, resilience to a changing social environment would prove adaptive for both surviving kin and non-kin.

Central distribution of oxytocin and vasopressin 1a receptors in juvenile Richardson's ground squirrels.

Oxytocin and vasopressin are well-conserved peptides important to the regulation of numerous aspects of social behavior, including sociality. Research exploring the distribution of the receptors for oxytocin (Oxtr) and for vasopressin (Avpr1a) in mammals has revealed associations between receptor distribution, sociality, and species' mating systems. Given that sociality and gregariousness can be tightly linked to reproduction, these nonapeptides unsurprisingly support affiliative behaviors that are important for mating and offspring care. We localized these receptors in juvenile Richardson's ground squirrel brains to determine whether distribution patterns of Oxtr and Avpr1a that are associated with promiscuous mating systems differ in rodents that also exhibit non-reproductive affiliation. These squirrels are social, colonial, and engage in nepotistic alarm calling behavior and affiliation outside of a reproductive context. Juveniles are the most affiliative age-class and are non-reproductive; making them ideal for examining these associations. We found that juveniles had dense Oxtr binding in the dentate gyrus of the hippocampus, amygdala, lateral septum, bed nucleus of the stria terminalis and medial geniculate nucleus. Juveniles had low to modest levels of Avpr1a binding in the medial preoptic area, olfactory bulbs, nucleus accumbens, superior colliculus, and inferior colliculus. We noted Oxtr and Avpr1a binding in the social behavior neural network (SBNN), further supporting a role of these nonapeptides in modulating social behavior across taxa. Oxtr and Avpr1a binding was also present in brain regions important to auditory processing that have known projections to the SBNN. We speculate that these neural substrates may be where these nonapeptides regulate communication.

Like mother, like daughter: heritability of female Richardson's ground squirrel Urocitellus richardsonii cortisol stress responses.

Activation of the hypothalamic-pituitary-adrenal (HPA) axis liberates glucocorticoids, which provides an acute indication of an individual's response to stressors. The heritability of the stress response in wild mammals, however, remains poorly documented. We quantified the cortisol stress response of female Richardson's ground squirrels (RGSs) to handling and physical restraint, testing for: (1) the effects of individual age, time of day, and sample latency; (2) repeatability within individuals; (3) narrow-sense heritability; and (4) differences among individuals owing to potential genetic and/or environmental effects. We detected a positive linear relationship between baseline plasma cortisol (BL-cortisol) concentration and stress-induced plasma cortisol (SI-cortisol) concentration that defined each individual's cortisol stress response. BL-cortisol, SI-cortisol, and stress response did not differ according to the time the sample was taken, or by subject age. Cortisol stress response was highly repeatable within individuals, had a mother-offspring heritability of h 2 = 0.40 ± 0.24 (mean ± SE), full-sibling heritability of h FS 2 = 0.37 ± 0.71 , and half-sibling heritability of h HS 2 = 0.75 ± 1.41 . Stress responses of sibling groups, immediate-family groups, and squirrels within a given area did not differ, whereas those of individuals from more distantly related matrilines did. Our results highlight the natural variability in HPA axis reactivity among individuals by quantifying both BL- and SI-cortisol levels, demonstrate partial heritability of the stress response that is not attributable to environmental variation, and suggest that at least part of an individual's stress response can be accounted for by differences in matrilineal history.

Effects of arginine vasopressin on Richardson's ground squirrel social and vocal behavior.

In nearly every vertebrate species examined thus far arginine vasopressin (AVP) and its homologues modulate behavior; thus, providing rich systems for comparative research. In rodents, AVP is best known for its modulation of social behavior; however, to date, research on AVPs effects on behavior have been limited to laboratory models and a few experiments using large outdoor enclosures. To extend our understanding of AVPs role in modulating social behavior and communication in an ecologically relevant context, we examined the effects of AVP on behavior of free-living Richardson's ground squirrels (Urocitellus richardsonii). To test the hypothesis that AVP influences social behavior and communication, we implanted osmotic minipumps into Richardson's ground squirrels and centrally administered AVP or saline as a control. Three different behavioral experiments quantifying behavior before and after AVP or saline administration were performed: a general behavior survey, a predator model presentation, and a social challenge test. AVP administration increased male vocalization rate when approached by a conspecific, but not when presented with a predator model. In males, social aggression decreased, but antipredator vigilance increased with AVP administration. Finally, AVP-treated females had increased "anxiety-like" behaviors during the social challenge test. Our data reveal that AVP has sex-specific effects on vocalizations and antipredator vigilance, as well as other social behaviors. Further, our data illustrate the importance of social context to AVPs modulation of behavior. (PsycINFO Database Record

Oxytocin and behavior: Lessons from knockout mice.

It is well established that the nonapeptide oxytocin (Oxt) is important for the neural modulation of behaviors in many mammalian species. Since its discovery in 1906 and synthesis in the early 1950s, elegant pharmacological work has helped identify specific neural substrates on which Oxt exerts its effects. More recently, mice with targeted genetic disruptions of the Oxt system-i.e., both the peptide and its receptor (the Oxtr)-have further defined Oxt's actions and laid some important scientific groundwork for studies in other species. In this article, we highlight the scientific contributions that various mouse knockouts of the Oxt system have made to our understanding of Oxt's modulation of behavior. We specifically focus on how the use of these mice has shed light on our understanding of social recognition memory, maternal behavior, aggression, and several nonsocial behaviors. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 77: 190-201, 2017.