Monogamous California Mouse Research Articles

Effects of reproductive status on behavioral and neural responses to isolated pup stimuli in female California mice

The transition to motherhood in mammals is marked by changes in females’ perception of and responsiveness to sensory stimuli from infants. Our understanding of maternally induced sensory plasticity relies most heavily on studies in uniparental, promiscuous house mice and rats, which may not be representative of rodent species with different life histories. We exposed biparental, monogamous California mouse (Peromyscus californicus) mothers and ovariectomized virgin females to one of four acoustic and olfactory stimulus combinations (Control: clean cotton and white noise; Call: clean cotton and pup vocalizations; Odor: pup-scented cotton and white noise; Call + Odor: pup-scented cotton and pup vocalizations) and quantified females’ behavior and Fos expression in select brain regions. Behavior did not differ between mothers and ovariectomized virgins. Among mothers, however, those exposed to the Control condition took the longest to sniff the odor stimulus, and mothers exposed to the Odor condition were quicker to sniff the odor ball compared to those in the Call condition. Behavior did not differ among ovariectomized virgins exposed to the different conditions. Fos expression differed across conditions only in the anterior hypothalamic nucleus (AHN), which responds to aversive stimuli: among mothers, the Control condition elicited the highest AHN Fos and Call + Odor elicited the lowest. Among ovariectomized virgin females, Call elicited the lowest Fos in the AHN. Thus, reproductive status in California mice alters females’ behavioral responses to stimuli from pups, especially odors, and results in the inhibition of defense circuitry in response to pup stimuli.

Testosterone pulses paired with a location induce a place preference to the nest of a monogamous mouse under field conditions.

Changing social environments such as the birth of young or aggressive encounters present a need to adjust behavior. Previous research examined how long-term changes in steroid hormones mediate these adjustments. We tested the novel concept that the rewarding effects of transient testosterone pulses (T-pulses) in males after social encounters alter their spatial distribution on a territory. In free-living monogamous California mice (Peromyscus californicus), males administered three T-injections at the nest spent more time at the nest than males treated with placebo injections. This mimics T-induced place preferences in the laboratory. Female mates of T-treated males spent less time at the nest but the pair produced more vocalizations and call types than controls. Traditionally, transient T-changes were thought to have transient behavioral effects. Our work demonstrates that in the wild, when T-pulses occur in a salient context such as a territory, the behavioral effects last days after T-levels return to baseline.

Reproductive success diminished following mate loss for females but not males in a monogamous rodent

The strictly monogamous California mouse (Peromyscus californicus) forms life-long pair bonds and mates exclusively with a single partner. While studies in the wild indicate that individuals may re-pair with a new partner following mate loss, the preponderance of this behavior and subsequent reproductive outcomes following re-pairing are understudied. To examine reproductive outcomes following re-pairing and to look for sex-specific differences following mate loss, birth records of 584 California mouse pairs from our laboratory were analyzed. Of these pairs, 59 pairs were identified as re-pairs and used for further descriptive analysis. We found that 50/59 (84.7 %) of re-paired animals gave birth, indicating that reproduction with a new mate is not only possible, but perhaps more common than previously described for this species. Additionally, we found that when re-paired, females took significantly longer to birth a subsequent litter as compared to original breeding pairs. Overall findings from the current study provide evidence for sex differences in reproductive outcomes following repairing and for greater flexibility in mating strategy for a species described as strictly monogamous.

Mating system is correlated with immunogenetic diversity in sympatric species of Peromyscine mice.

The number of reproductive partners per individual varies markedly across animal mating systems. This variation may be an important determinant of patterns of immunogenetic diversity, particularly at Major Histocompatibility Complex (MHC) Class I and II loci. To compare immunogenetic variation in taxa with markedly different mating systems, we used RNAseq-generated data to quantify genotypic diversity in three species of Peromyscine rodents: the monogamous California mouse (Peromyscus californicus) and the polygynandrous deer mouse (P. maniculatus) and brush mouse (P. boylii). By sampling populations of these species from multiple localities in California, we were able to conduct replicated analyses of the relationship between mating system and immunogenetic variation. Across the four localities sampled, diversity at MHC Class I and II genes was consistently higher in the two polygynandrous species. We found no evidence that sampling location (i.e., variation in habitat conditions) contributed to observed differences in MHC variation among populations or species. Collectively, our data indicate that immunogenetic variation in Peromyscine mice is associated with reproductive behavior, rather than geographic locality or habitat type. The consistently greater variability detected in the polygynandrous species examined suggests that balancing selection imposed by behaviorally-mediated pathogen exposure is important in maintaining variation at MHC genes in these animals.

Rapid effects of testosterone on social decision-making in a monogamous California mice (Peromyscus californicus)

Social animals must cope with challenges and opportunities by adjusting how they react to a salient stimulus. Here we use California mice (Peromyscus californicus) and investigate the mechanisms underlying social decision-making by studying (i) rapid effects of testosterone (T) pulses on a male’s decisions to approach a novel male (challenge) versus a receptive female (opportunity), and (ii) whether social experience shapes how such effects are manifested. In Experiment 1, we found that sexually naïve males administered saline injections preferentially approached unfamiliar females over unfamiliar males, in contrast, 10 min after receiving a single T-injection, males expressed a preference for approaching unfamiliar males. Such an effect of T only occurred in sexually naïve males, but not pair-bonded males, suggesting that the rapid effects of T on approach behavior may rely on the pair-bonding experiences. Experiment 2 investigated social decision-making across three repeated exposures to the challenge/opportunity situations. Only the initial decision, approach to the challenge, predicted future aggressive behaviors, and such an effect relied on the rapid actions of T. We also found that experience with the controlled challenge situation (the male intruder was restrained behind a wire mesh) dampened the approach to the male side (potential threat) when later exposed to the same conditions. This suggests that a resident’s motivation to defend against a threatening individual may decrease as the threat posed by the “neighbors” is reduced. Overall rapid effects of post-encounter T pulses may play important roles in influencing behavioral decisions during social interactions.

Differential gene expression in relation to mating system in Peromyscine rodents.

Behaviors that increase an individual's exposure to pathogens are expected to have important effects on immunoactivity. Because sexual reproduction typically requires close contact among conspecifics, mating systems provide an ideal opportunity to study the immunogenetic correlates of behaviors with high versus low risks of pathogen exposure. Despite logical links between polygynandrous mating behavior, increased pathogen exposure, and greater immunoactivity, these relationships have seldom been examined in nonhuman vertebrates. To explore interactions among these variables in a different lineage of mammals, we used RNAseq to study the gene expression profiles of liver tissue—a highly immunoactive organ—from sympatric populations of the monogamous California mouse (Peromyscus californicus) and two polygynandrous congeners (P. maniculatus and P. boylii). Differential expression and co‐expression analyses revealed distinct patterns of gene activity among species, with much of this variation associated with differences in mating system. This tendency was particularly pronounced for MHC genes, with multiple MHC Class I genes being upregulated in the two polygynandrous species, as expected if exposure to sexually transmitted pathogens varies with mating system. Our results underscore the role of mating behavior in influencing patterns of gene expression and highlight the use of emerging transcriptomic tools in behavioral studies of free‐living animals.

Changes in Behavior and Ultrasonic Vocalizations During Pair Bonding and in Response to an Infidelity Challenge in Monogamous California Mice

Despite recent exciting research about pair bonding, little is known about how mammalian vocalizations change with the initiation and maintenance of pair bonding in monogamous species. Moreover, even less is known about the significance of pair bond resilience in the face of social challenges. In the strictly monogamous California mouse (Peromyscus californicus), we measured changes in ultrasonic vocalizations (USV) and other behaviors within male-female dyads over the course of pair bonding and characterized associations of USVs with affiliation and aggression. After one week of cohabitation, pairs exhibited decreased aggression and “bark” USVs, and increased “simple sweep” and “sustained vocalization” (SV) USV types. Accordingly, the number of barks was associated with aggression, whereas the number of simple sweeps and the number, call duration and bout size of SVs corresponded with affiliation. We then experimentally assessed the impact of an infidelity challenge (one week cohabitation with an unfamiliar, opposite-sex, extra-pair individual) for both sexes on pair social behavior, acoustic behavior, and reproductive success. The infidelity challenge temporarily disrupted pair bond interactions during pair reunion, independent of which sex experienced the infidelity challenge, via both increases in aggression and barks, and a stunting of affiliation and SVs, compared to control pairs. Pair reproductive success, in the form of birth latency, litter size, pup survival and birth weight, did not differ between infidelity challenge pairs and controls. The quality of pair interactions, however, was associated with reproductive success: aggression during pair reunion across all pairs was associated with a lower likelihood of successfully producing a litter. Similarly, among infidelity challenge pairs, but not the controls, there was a positive association between pair affiliation and paternal care, and a negative association between pair aggression and paternal care. Overall, the infidelity challenge revealed a weak negative effect on reproductive success, but we speculate, based on our results, that greater resiliency of a pair bond can moderate negative effects of a social challenge.

Genomic analysis of MHC-based mate choice in the monogamous California mouse.

Variation at Major Histocompatibility Complex (MHC) genes is thought to be an important mechanism underlying mate choice in vertebrates, with individuals typically predicted to prefer MHC-dissimilar reproductive partners. However, analyses based on individual MHC loci have generated contradictory results regarding the role of these genes in mate-choice decisions. To provide a more comprehensive assessment of relationships between MHC variation and mating behavior, we used an exome capture strategy to characterize variability at 13 MHC loci, 312 innate immune system genes, and 1044 nonimmune genes in 25 obligate monogamous pairs of California mice (Peromyscus californicus) from 2 free-living populations of this species in Monterey County, California. Pairwise genotypic comparisons and analyses of SNP-based allelic differences failed to detect disassortative mating based on MHC variability; reproductive partners were not more dissimilar than randomly generated male-female pairs at MHC, innate or nonimmune loci. Within populations, individuals tended to be more closely related at MHC genes than at innate or nonimmune genes. Consistent with the functional role of immunogenes, the 2 study populations were highly differentiated at MHC and innate genes but not at nonimmune loci. Collectively, our results suggest that MHC genetic variation in California mice reflects local differences in pathogen exposure rather than disassortative mating based on variability at MHC Class I and II genes.

Effects of social defeat on paternal behavior and pair bonding behavior in male California mice (Peromyscus californicus)

Male parental care is an important social behavior for several mammalian species. Psychosocial stress is usually found to inhibit maternal behavior, but effects on paternal behavior have been less consistent. We tested the effects of social defeat stress on pair bond formation and paternal behavior in the monogamous California mouse (Peromyscus californicus). Social defeat reduced time spent in a chamber with a stranger female during a partner preference test conducted 24h after pairing, but increased latency to the first litter. In 10min partner preference tests conducted after the birth of pups, both control and stressed males exhibited selective aggression towards stranger females. Unlike prairie voles, side by side contact was not observed in either partner preference test. Stressed male California mice engaged in more paternal behavior than controls and had reduced anxiety-like responses in the open-field test. Defeat stress enhanced prodynorphin and KOR expression in the medial preoptic area (MPOA) but not PVN. Increased KOR signaling has been linked to increased selective aggression in prairie voles. Together the results show that defeat stress enhances behaviors related to parental care and pair bonding in male California mice.

Ultrasonic vocalization production and playback predicts intrapair and extrapair social behaviour in a monogamous mouse

Most studies examining rodent ultrasonic vocalizations (USVs) have investigated pericopulatory vocal behaviour in polygynous rodents, while vocalizations related to pair bond maintenance in monogamous rodents remain unexplored. In the monogamous California mouse, Peromyscus californicus, we used ultrasonic playbacks and post-playback social interactions to assess possible functions of USVs. We found that females responded with approach towards USVs of an unfamiliar male (bonded male from another pair) compared to a noise control, but displayed no difference in response to calls of their partner versus noise. Responsiveness to unfamiliar males does not appear to reflect an interest in extrapair copulations because during post-playback social interactions, females displayed more agonistic behaviours and fewer affiliative behaviours towards unfamiliar, sexually naive ‘stranger’ males than towards their partners. We speculate that approach to unfamiliar male USVs instead may be related to territorial defence. We further explored associations within the data set. Interestingly, female affiliation with her partner was predicted by USV output, particularly a higher number and proportion of complex call types, produced during the male partner USV elicitation phase. Female approach towards USVs was related to syllable duration of one call type in partner USVs (not in unfamiliar USVs) but no other features, and sufficient variation exists in syllable duration to allow females to theoretically distinguish between individuals based on this measure. Similarly, while pregnancy state did not influence female social behaviour, it decreased approach to playback of partner USVs but not to that of unfamiliar USVs. Overall, our results illuminate concepts about vocal communication in monogamous rodent species with strong pair bonds and suggest that functions of USVs in rodents can extend beyond mate choice.

Effects of social defeat on dopamine neurons in the ventral tegmental area in male and female California mice.

Dopamine neurons in the ventral tegmental area (VTA) have important functions related to rewards but are also activated in aversive contexts. Electrophysiology studies suggest that the degree to which VTA dopamine neurons respond to noxious stimuli is topographically organized across the dorsal-ventral extent. We used c-fos immunohistochemistry to examine the responses of VTA dopamine neurons in contexts of social defeat and social approach. Studying monogamous California mice (Peromyscus californicus) allowed us to observe the effects of social defeat on both males and females. Females exposed to three episodes of defeat, but not a single episode, had more tyrosine hydroxylase (TH)/c-fos-positive cells in the ventral (but not dorsal) VTA compared with controls. This observation suggests that repeated exposure to aversive contexts is necessary to trigger activation of VTA dopamine neurons. Defeat did not affect TH/c-fos colocalizations in males. We also examined the long-term effects of defeat on c-fos expression in a social interaction test. As previously reported, defeat reduced social interaction in females but not males. Surprisingly, there were no effects of defeat stress on TH/c-fos colocalizations in any subregion of the VTA. However, females had more TH/c-fos-positive cells than males across the entire VTA, and also had greater c-fos-positive cell counts in posterior subregions of the nucleus accumbens shell. Our results show that dopamine neurons in the VTA are more responsive to social contexts in females and that the ventral VTA in particular is sensitive to aversive contexts.

Male fidelity expressed through rapid testosterone suppression of ultrasonic vocalizations to novel females in the monogamous California mouse

The steroid hormone testosterone (T) is a well-known mediator of male sexual behavior in vertebrates. However, less is known about T's rapid effects on sexual behavior, particularly those involving ultrasonic vocalizations (USVs), a mode of communication that can influence mate acquisition in rodents. Using the monogamous California mouse, Peromyscus californicus, we tested whether T rapidly alters male USV production by giving T or saline injections to non-paired (sexually naïve) males and paired (paternally experienced and pair-bonded) males immediately prior to a brief exposure to an unrelated, novel female. Among non-paired males, no differences in the total number of USVs were observed; however, T increased the proportion of simple sweeps produced. Among paired males, T decreased the number of USVs produced, and this change was driven by a reduction in simple sweeps. These results suggest a differential rapid effect of T pulses between non-paired and paired males upon exposure to a novel female. Additionally, we observed a positive correlation in the production of USVs made between males and novel females, and this relationship was altered by T. Given the importance of USVs in sexual communication, our study supports an essential concept of monogamy in that mate fidelity is reinforced by decreased responsiveness to prospective mates outside of the pair bond. The central mechanism in pair bonded males that decreases their responsiveness to novel females appears to be one that T can trigger. This is among the first studies to demonstrate that T can inhibit sexually related behaviors and do so rapidly.

Sex differences in stress-induced social withdrawal: role of brain derived neurotrophic factor in the bed nucleus of the stria terminalis

Depression and anxiety disorders are more common in women than men, and little is known about the neurobiological mechanisms that contribute to this disparity. Recent data suggest that stress-induced changes in neurotrophins have opposing effects on behavior by acting in different brain networks. Social defeat has been an important approach for understanding neurotrophin action, but low female aggression levels in rats and mice have limited the application of these methods primarily to males. We examined the effects of social defeat in monogamous California mice (Peromyscus californicus), a species in which both males and females defend territories. We demonstrate that defeat stress increases mature brain-derived neurotrophic factor (BDNF) protein but not mRNA in the bed nucleus of the stria terminalis (BNST) in females but not males. Changes in BDNF protein were limited to anterior subregions of the BNST, and there were no changes in the adjacent nucleus accumbens (NAc). The effects of defeat on social withdrawal behavior and BDNF were reversed by chronic, low doses of the antidepressant sertraline. However, higher doses of sertraline restored social withdrawal and elevated BDNF levels. Acute treatment with a low dose of sertraline failed to reverse the effects of defeat. Infusions of the selective tyrosine-related kinase B receptor (TrkB) antagonist ANA-12 into the anterior BNST specifically increased social interaction in stressed females but had no effect on behavior in females naïve to defeat. These results suggest that stress-induced increases in BDNF in the anterior BNST contribute to the exaggerated social withdrawal phenotype observed in females.

Father Absence in the Monogamous California Mouse Impairs Social Behavior and Modifies Dopamine and Glutamate Synapses in the Medial Prefrontal Cortex

The role of the father in psycho-affective development is indispensable. Yet, the neurobehavioral effects of paternal deprivation (PD) are poorly understood. Here, we examined the behavioral consequences of PD in the California mouse, a species displaying monogamous bonding and biparental care, and assessed its impact on dopamine (DA), serotonin (5-HT), and glutamate (GLU) transmission in the medial prefrontal cortex (mPFC). In adult males, deficits in social interaction were observed, when a father-deprived (PD) mouse was matched with a PD partner. In adult females, deficits were observed when matching a PD animal with a non-PD control, and when matching 2 PD animals. PD also increased aggression in females. Behavioral abnormalities in PD females were associated with a sensitized response to the locomotor-activating effect of amphetamine. Following immunocytochemical demonstration of DA, 5-HT, and GLU innervations in the mPFC, we employed in vivo electrophysiology and microiontophoresis, and found that PD attenuated the basal activity of low-spiking pyramidal neurons in females. PD decreased pyramidal responses to DA in females, while enhancing responses to NMDA in both sexes. We thus demonstrate that, during critical neurodevelopmental periods, PD leads to sex-dependent abnormalities in social and reward-related behaviors that are associated with disturbances in cortical DA and GLU neurotransmission.

Sex differences in effects of dopamine D1 receptors on social withdrawal

Dopamine signaling in the nucleus accumbens (NAc) plays a critical role in the regulation of motivational states. Recent studies in male rodents show that social defeat stress increases the activity of ventral tegmental dopamine neurons projecting to the NAc, and that this increased activity is necessary for stress-induced social withdrawal. Domestic female mice are not similarly aggressive, which has hindered complementary studies in females. Using the monogamous California mouse (Peromyscus californicus), we found that social defeat increased total dopamine, DOPAC, and HVA content in the NAc in both males and females. These results are generally consistent with previous studies in Mus, and suggest defeat stress also increases NAc dopamine signaling in females. However, these results do not explain our previous observations that defeat stress induces social withdrawal in female but not male California mice. Pharmacological manipulations provided more insights. When 500 ng of the D1 agonist SKF38393 was infused in the NAc shell of females that were naïve to defeat, social interaction behavior was reduced. This same dose of SKF38393 had no effect in males, suggesting that D1 receptor activation is sufficient to induce social withdrawal in females but not males. Intra-accumbens infusion of the D1 antagonist SCH23390 increased social approach behavior in females exposed to defeat but not in females naïve to defeat. This result suggests that D1 receptors are necessary for defeat-induced social withdrawal. Overall, our results suggest that sex differences in molecular pathways that are regulated by D1 receptors contribute to sex differences in social withdrawal behavior.

Interactions between parents and parents and pups in the monogamous California mouse (Peromyscus californicus).

The California mouse ( Peromyscus californicus ) may be a valuable animal model to study parenting as it is one of the few monogamous and biparental rodent species. By using automated infra-red imaging and video documentation of established pairs spanning two days prior to birth of the litter until d 5 of post natal development (PND), it was possible to follow interactions between parents and between parents and pups. The paired males were attentive to their partners in the form of grooming and sniffing throughout the time period studied. Both these and other activities of the partners, such as eating and drinking, peaked during late light/ mid-dark period. Beginning the day before birth, and most significantly on PND 0, the female made aggressive attempts to exclude the male from nest-attending, acts that were not reciprocated by the male, although he made repeated attempts to mate his partner during that period. By PND 1, males were permitted to return to the nest, where they initiated grooming, licking, and huddling over the litter, although time spent by the male on parental care was still less than that of the female. Male and female pups were of similar size and grew at the same rate. Pups, which are believed to be exothermic for at least the first two weeks post-natally, maintained a body temperature higher than that of their parents until PND 16. Data are consistent with the inference that the male California mouse parent is important in helping retain pup body heat and permit dams increased time to procure food to accommodate her increased energy needs for lactation. These assessments provide indices that may be used to assess the effects of extrinsic factors, such as endocrine disrupting chemicals, on biparental behaviors and offspring development.

Non-genomic transmission of paternal behaviour between fathers and sons in the monogamous and biparental California mouse

Maternal behaviour has profound, long-lasting implications for the health and well-being of developing offspring. In the monogamous California mouse (Peromyscus californicus), care by both parents is critical for offspring survival. We tested the hypothesis that similar to maternal care in rodents, paternal huddling and grooming (HG) behaviour can be transmitted to future generations via behavioural mechanisms. In California mice, testosterone maintains paternal HG behaviour. In the present study, we randomly assigned a group of male California mice to castration or sham-operated conditions and allowed them to raise their offspring normally. Adult sons of these males were paired with a female, and they were observed interacting with their own offspring. We found that like their fathers, the sons of castrated males huddled and groomed their young at lower levels than the sons of sham-operated fathers. The sons of castrates also retrieved pups more frequently. When both parents were present, the sons of castrates also showed a trend towards engaging in less exploratory behaviour. These data support the hypothesis that paternal behaviour, like maternal behaviour, can be transferred to future generations via epigenetic mechanisms and suggest that in a biparental species both parents contribute to offspring behavioural development.

Sex differences in stress-induced social withdrawal: Independence from adult gonadal hormones and inhibition of female phenotype by corncob bedding

There is compelling evidence for important sex differences in behavioral and hormonal responses to psychosocial stress. Here we examined the effects of gonadal hormones on behavioral responses to social defeat stress in monogamous California mice (Peromyscus californicus). Three episodes of social defeat induced social withdrawal in intact females but not males. Gonadectomy blocked corticosterone responses to defeat in females and sensitized male corticosterone responses. However, gonadectomy had no effects on social interaction behavior, suggesting that social withdrawal is not dependent on gonadal hormones in the adult California mouse. In contrast, defeat reduced exploratory behavior in the open field test for intact but not castrated males. We also examined the effects of social defeat on social interaction behavior when California mice were raised on corncob bedding, which has estrogenic properties. In this dataset of over 300 mice, we observed that social defeat did not induce social withdrawal when females were raised on corncob bedding. This finding suggests that the use of corncob in rodent studies could mask important sex differences in the effects of stress on brain and behavior. Although gonadal hormones do not affect social withdrawal behavior in adults, our data suggest that hormones may act earlier in development to induce a more resilient social phenotype.

A positive Link Between Male Testosterone and Spacing Behavior in Pair‐Bonded California Mice

AbstractThe effects of testosterone (T) on male sexual behavior are well established, but its roles in several associated social behaviors such as pair‐bonding and paternal behavior are diverse. Recently, we reported that male T in response to pairing with a female predicts future paternal behavior in the monogamous and biparental California mouse (Peromyscus californicus). Here, we examined whether T in response to pairing is also associated with spacing behavior between mates under laboratory conditions and if these behavioral patterns are continued during pup care. Pairs of California mice were observed throughout their pair bond before and again after pups were born. We found that males with higher T post‐pairing remained closer to their mates in the weeks before pups were born, and pairs with higher proximity measures during the pair bond spent more time caring for pups together. Overall, these results suggest a similar mechanism for spacing behavior across distinct phases of the pair bond.

Monogamous and Promiscuous Rodent Species Exhibit Discrete Variation in the Size of the Medial Prefrontal Cortex

Limbic-associated cortical areas, such as the medial prefrontal and retrosplenial cortex (mPFC and RS, respectively), are involved in the processing of emotion, motivation, and various aspects of working memory and have been implicated in mating behavior. To determine whether the independent evolution of mating systems is associated with a convergence in cortical mechanisms, we compared the size of mPFC and RS between the monogamous prairie vole (Microtus ochrogaster) and the promiscuous meadow vole (Microtus pennsylvanicus), and between the monogamous California mouse (Peromyscus californicus) and the promiscuous white-footed mouse (Peromyscus leucopus). For both promiscuous mice and voles, the mPFC occupied a significantly larger percentage of total cortex than in the monogamous species. No significant differences were observed for the RS or overall cortex size with respect to mating system, supporting the convergent evolution of mPFC size, specifically. Individual differences in the mating behavior of male prairie voles (wandering versus pair-bonding), presumably facultative tactics, were not reflected in the relative size of the mPFC, which is likely a heritable trait. Given the importance of the mPFC for complex working memory, particularly object-place and temporal order memory, we hypothesize that the relatively greater size of the mPFC in promiscuous species reflects a greater need to remember multiple individuals and the times and locations in which they have been encountered in the home range.