Differences in Diffusion-Weighted Imaging and Resting-State Functional Connectivity Between Two Culturally Distinct Populations of Prairie Vole.

BACKGROUND: We used the highly prosocial prairie vole to test the hypothesis that higher-order brain structure-microarchitecture and functional connectivity (FC)-would differ between males from populations with distinctly different levels of prosocial behavior. Specifically, we studied males from Illinois (IL), which display high levels of prosocial behavior, and first generation males from Kansas dams and IL males (KI), which display the lowest level of prosocial behavior and higher aggression. Behavioral differences between these males are associated with overexpression of estrogen receptor alpha in the medial amygdala and bed nucleus of the stria terminalis and neuropeptide expression in the paraventricular nucleus. METHODS: We compared apparent diffusion coefficient, fractional anisotropy, and blood oxygen level-dependent resting-state FC between males. RESULTS: IL males displayed higher apparent diffusion coefficient in regions associated with prosocial behavior, including the bed nucleus of the stria terminalis, paraventricular nucleus, and anterior thalamic nuclei, while KI males showed higher apparent diffusion coefficient in the brainstem. KI males showed significantly higher fractional anisotropy than IL males in 26 brain regions, with the majority being in the brainstem reticular activating system. IL males showed more blood oxygen level-dependent resting-state FC between the bed nucleus of the stria terminalis, paraventricular nucleus, and medial amygdala along with other brain regions, including the hippocampus and areas associated with social and reward networks. CONCLUSIONS: Our results suggest that gray matter microarchitecture and FC may play a role the expression of prosocial behavior and that differences in other brain regions, especially the brainstem, could be involved. The differences between males suggests that this system represents a potentially valuable model system for studying emotional differences and vulnerability to stress and addiction.

Medial amygdala ERα expression influences monogamous behaviour of male prairie voles in the field.

Formation of long-term pair-bonds is a complex process, involving multiple neural circuits and is context- and experience-dependent. While laboratory studies using prairie voles have identified the involvement of several neural mechanisms, efforts to translate these findings into predictable field outcomes have been inconsistent at best. Here we test the hypothesis that inhibition of oestrogen receptor alpha (ERα) in the medial amygdala of male prairie voles would significantly increase the expression of social monogamy in the field. Prairie vole populations of equal sex ratio were established in outdoor enclosures with males bred for high levels of ERα expression and low levels of prosocial behaviour associated with social monogamy. Medial amygdala ERα expression was knocked down in half the males per population. Knockdown males displayed a greater degree of social monogamy in five of the eight behavioural indices assessed. This study demonstrates the robust nature of ERα in playing a critical role in the expression of male social monogamy in a field setting.

Breeding patterns of female prairie voles (Microtus ochrogaster) displaying alternative reproductive tactics.

Individuals of either sex may display alternative behaviors to obtain copulations, but few studies have examined the breeding patterns of females and males in populations where individuals of both sexes exhibit alternative reproductive tactics (ARTs). In prairie voles (Microtus ochrogaster), most adults are territorial, residing at a single nest site either as male-female pairs or as solitary individuals. However, some adults adopt nonterritorial, wandering tactics. During two field seasons monitoring prairie vole populations maintained in seminatural enclosures, we found evidence that females exhibiting different ARTs bred differentially with resident and wandering males. Females residing at a nest with a male bred significantly more often with a paired resident male, primarily their social partner, and significantly less often with male wanderers compared to single resident females or wandering females. These patterns were not due to chance, because paired resident females produced offspring with paired resident males significantly more than expected based on the relative abundance of these males in the population, whereas single resident females produced offspring with male wanderers significantly more than expected based on the proportion of male wanderers in the population. We did not find any evidence that multiple paternity was greater in the litters of single resident females and wanderer females even though these females lacked a male social partner to limit mating access by multiple males. This suggests that mate guarding by a female's male social partner was not the primary determinant of multiple paternity in the litters of females exhibiting different reproductive tactics. However, male ART did affect the likelihood of multiple paternity. Females that produced offspring with single resident or wanderer males had an increased likelihood of multiple paternity relative to females producing offspring with paired resident males. The results of this study show that female and male ARTs can affect breeding patterns.

Do olfactory cues from males with different avpr1a genotypes affect female mate choice in prairie voles, Microtus ochrogaster?

Multiple hypotheses have been proposed to explain female mate choice and some of the mechanisms underlying these choices. Females prairie voles display social and mating preferences for males with longer avpr1a microsatellite alleles, which provide more paternal care and exhibit less interest in novel females compared to males with shorter avpr1a microsatellite alleles. The cues females use to differentiate among males with different avpr1a genotypes are unknown, so the objective of our study was to determine if females can discriminate among males with different avpr1a genotypes using only male olfactory cues. In a laboratory choice test, females simultaneously presented with soiled bedding from a male with short versus long avpr1a microsatellite alleles showed no significant difference in the total time spent investigating each type of bedding. Nor did a greater number of females spend more time investigating soiled bedding from males with short versus long avpr1a microsatellite alleles. These findings were not influenced by female estrous status or their own avpr1a genotype. Our results suggest olfactory cues alone are insufficient to explain a female's ability to discriminate between males with different avpr1a genotypes and future research should focus on different cues or a combination of cues.

Male prairie voles with different avpr1a microsatellite lengths do not differ in courtship behaviour.

Females are generally expected to be selective when choosing their social and sexual partners. In a previous laboratory study, female prairie voles (Microtus ochrogaster) showed significant social and sexual preferences for males with longer microsatellite DNA within the avpr1a gene encoding the vasopressin 1a receptor, as predicted if females select mates whose parental behaviour should increase female reproductive success. We tested the hypothesis that males with short versus long avpr1a microsatellite alleles exhibit differences in courtship behaviour, which could act as cues for female mate preference. The only behavioural difference we detected between males with short versus long avpr1a microsatellite alleles in mate preference trials was that males with short avpr1a microsatellite alleles sniffed the anogenital region of females more frequently during the first two days of the trials. Our results did not strongly support the hypothesis that a male's avpr1a genotype predicts the courtship behaviours we measured and suggests that other courtship behaviours or traits, such as odour and vocalizations, may be more important to female prairie voles when choosing mates. Additional studies using a wider array of species are needed to assess the degree to which male mammal courtship behaviour provides information on mate quality to females.

Taxon matters: promoting integrative studies of social behavior: NESCent Working Group on Integrative Models of Vertebrate Sociality: Evolution, Mechanisms, and Emergent Properties.

The neural and molecular mechanisms underlying social behavior - including their functional significance and evolution - can only be fully understood using data obtained under multiple social, environmental, and physiological conditions. Understanding the complexity of social behavior requires integration across levels of analysis in both laboratory and field settings. However, there is currently a disconnect between the systems studied in the laboratory versus the field. We argue that recent conceptual and technical advances provide exciting new opportunities to close this gap by making non-model organisms accessible to modern approaches in both laboratory and nature.

Fine-scale spatial patterns of genetic relatedness among resident adult prairie voles.

Characterizing the spatial arrangement of related individuals within populations can convey information about opportunities for the evolution of kin-selected social behaviors, the potential for inbreeding, and the geographic distribution of genetic variation. Prairie voles (Microtus ochrogaster) are socially monogamous rodents that sometimes breed cooperatively. Individuals of both sexes are highly philopatric, and among natal dispersers, the average dispersal distance is about 30 m. Such limited natal dispersal can result in the spatial clustering of kin and we used microsatellite data to estimate genetic relatedness among resident adult prairie voles in 2 natural populations to test the hypothesis that limited natal dispersal of male and female prairie voles results in the spatial clustering of kin. Spatial autocorrelation analyses of nest residency and microsatellite data indicated that proximate same-sex adult residents of both sexes were significantly more related than more spatially distant resident same-sex adults in Kansas. In Indiana, adult female voles residing less than 20 m apart were also significantly more related than more spatially distant resident adult females but spatial clustering of kin was not detected among resident adult males. The spatial clustering of kin indicates that opportunities for kin-selected behaviors exist in both populations, especially among females. Differences in the patterns of spatial genetic structure among resident males between the Kansas and Indiana populations may be due to population differences in factors such as demography and mating system, as well as in the extent of natal philopatry.

An evolutionary framework for studying mechanisms of social behavior.

Social interactions are central to most animals and have a fundamental impact upon the phenotype of an individual. Social behavior (social interactions among conspecifics) represents a central challenge to the integration of the functional and mechanistic bases of complex behavior. Traditionally, studies of proximate and ultimate elements of social behavior have been conducted by distinct groups of researchers, with little communication across perceived disciplinary boundaries. However, recent technological advances, coupled with increased recognition of the substantial variation in mechanisms underlying social interactions, should compel investigators from divergent disciplines to pursue more integrative analyses of social behavior. We propose an integrative conceptual framework intended to guide researchers towards a comprehensive understanding of the evolution and maintenance of mechanisms governing variation in sociality.

avpr1a length polymorphism is not associated with either social or genetic monogamy in free-living prairie voles.

Recent discoveries of single-gene influences on social behaviour have generated a great deal of interest in the proximate mechanisms underlying the expression of complex behaviours. Length polymorphism in a microsatellite in the regulatory region of the gene encoding the vasopressin 1a receptor (avpr1a) has been associated with both inter- and intra-specific variation in socially monogamous behaviour in voles (genus Microtus) under laboratory conditions. Here, we evaluate the relationship between avpr1a length polymorphism and social associations, genetic monogamy, and reproductive success in free-living prairie vole (M. ochrogaster) populations. We found no evidence of a relationship between avpr1a microsatellite length and any of our correlates of either social or genetic monogamy in the field. Our results, especially when taken in conjunction with those of recent experimental studies in semi-natural enclosures, suggest that avpr1a polymorphism is unlikely to have been a major influence in the evolution or maintenance of social monogamy in prairie voles under natural conditions.

Does time after pair bond disruption affect subsequent reproduction in the socially monogamous woodland vole (Microtus pinetorum)?

Disruption of the pair bond between socially monogamous animals leads to changes in behavior, which may have reproductive consequences. There are two alternative hypotheses to explain the effect of the length of time since pair bond disruption on subsequent reproduction. One hypothesis predicts that voles housed immediately with a new opposite-sex conspecific will be as likely to produce litters and will produce them as quickly as voles separated from their initial mate for longer. Alternatively, if attachment between mates is enduring, we expect that more voles separated longer from their previous mates will produce litters and produce them sooner than voles re-paired immediately after separation from their initial mates. Woodland voles, paired with opposite-sex conspecifics, remained together until parturition. Mates were then separated for 0, 7, or 14 days until re-pairing with an opposite-sex conspecific. Pair bond disruption did not prevent males and females from mating subsequently, which was consistent with data from our breeding colony. In addition, the length of time an individual remained alone after pair bond disruption did not affect the latency to produce a litter. Our results show that having been paired previously does not affect subsequent reproduction in this socially monogamous vole.

The relationship between dominance rank and spatial ability among male meadow voles (Microtus pennsylvanicus).

Males of many mammalian species exhibit contest competition and scramble competition for mates, but the relationship between these 2 forms of competition remains poorly understood. The authors measured dominance rank and spatial ability as traits likely to be selected by contest and scramble competition, respectively, among male meadow voles (Microtus pennsylvanicus). The spatial ability of males was assessed using water maze tests, and dominance rank was determined using paired trials in a neutral arena. Dominant males had better spatial-learning ability and tended to have quicker learning speed but did not have better spatial memory than less aggressive subordinates. Therefore, the authors found no evidence that contest and scramble competition have favored alternative reproductive phenotypes among male meadow voles.

Reproductive activation of pine voles (Microtus pinetorum): examination of physiological markers.

We tested the hypothesis that the presence of an opposite-sex conspecific will result in time-related changes in measures of reproductive activation. We housed male-female pairs of pine voles together for 0, 2, 6, 12, or 24 h before collecting blood, reproductive organs and brains for immunocytochemical analysis of LHRH and c-fos. Control animals were never exposed to an opposite-sex conspecific. Following exposure to a male, there was a significant increase in uterine weight but not in LH levels. In males, there were no changes in peripheral indices of activation, i.e. LH levels, testes and seminal vesicle weights were not altered. Consistent with no change in circulating levels of LH, there was no change in LHRH immunoreactivity at any time. However, c-fos immunoreactivity was significantly greater in both males and females in the cingulate cortex and rostral bed nucleus of the stria terminalis (BNST) at 2 h, and in the caudal BNST at 2, 6 and 12 h. Similarly, c-fos immunoreactivity was increased in the rostral MPOA in both males and females at 2 and 6 h. However, in the caudal MPOA, there was a significant interaction between sex and time due to increased c-fos immunoreactivity in females only at 6 h. These results indicate that, in both male and female pine voles, exposure to an opposite-sex conspecific is sufficient to produce rapid, neural activation in brain areas known to be involved in reproductive activation and sexual behavior. This early activation did not occur in LHRH neurons. It is not known if this activation, particularly at early times, is due to reproductive activation or to the formation of pair bonds.

Social transmission and memory of food preferences in pine voles (Microtus pinetorum).

The social transmission of food preferences is affected by factors including the length of time a preference is maintained. The authors investigated the social transmission of food preferences in pine voles (Microtus pinetorum) and whether food items had to be present for memory to persist. A demonstrator vole was fed cocoa-flavored food, a nonpreferred food. After interacting with a demonstrator vole, observer voles preferred cocoa-flavored food as compared with either naive voles that had not interacted with demonstrators or voles that had been exposed to odors. Observers retained this preference for 72 hr after interacting with an observer whether or not flavored foods were present. The ability to learn which food items are palatable and safe may benefit a generalist herbivore that has faced selection to avoid toxic baits.