Genetic variation in the developmental regulation of cortical avpr1a among prairie voles.

Early experiences can have enduring impacts on brain and behavior, but the strength of these effects can be influenced by genetic variation. In principle, polymorphic CpGs (polyCpGs) may contribute to gene-by-environment interactions (G × E) by altering DNA methylation. In this study, we investigate the influence of polyCpGs on the development of vasopressin receptor 1a abundance in the retrosplenial cortex (RSC-V1aR) of prairie voles (Microtus ochrogaster). Two alternative alleles ('HI'/'LO') predict RSC avpr1a expression, V1aR abundance and sexual fidelity in adulthood; these alleles differ in the frequency of CpG sites and in methylation at a putative intron enhancer. We hypothesized that the elevated CpG abundance in the LO allele would make homozygous LO/LO voles more sensitive to developmental perturbations. We found that genotype differences in RSC-V1aR abundance emerged early in ontogeny and were accompanied by differences in methylation of the putative enhancer. As predicted, postnatal treatment with an oxytocin receptor antagonist (OTA) reduced RSC-V1aR abundance in LO/LO adults but not their HI/HI siblings. Similarly, methylation inhibition by zebularine increased RSC-V1aR in LO/LO adults, but not in HI/HI siblings. These data show a gene-by-environment interaction in RSC-V1aR. Surprisingly, however, neither OTA nor zebularine altered adult methylation of the intronic enhancer, suggesting that differences in sensitivity could not be explained by CpG density at the enhancer alone. Methylated DNA immunoprecipiation-sequencing showed additional differentially methylated regions between HI/HI and LO/LO voles. Future research should examine the role of these regions and other regulatory elements in the ontogeny of RSC-V1aR and its developmentally induced changes.

Methylation of avpr1a in the cortex of wild prairie voles: effects of CpG position and polymorphism.

DNA methylation can cause stable changes in neuronal gene expression, but we know little about its role in individual differences in the wild. In this study, we focus on the vasopressin 1a receptor (avpr1a), a gene extensively implicated in vertebrate social behaviour, and explore natural variation in DNA methylation, genetic polymorphism and neuronal gene expression among 30 wild prairie voles (Microtus ochrogaster). Examination of CpG density across 8 kb of the locus revealed two distinct CpG islands overlapping promoter and first exon, characterized by few CpG polymorphisms. We used a targeted bisulfite sequencing approach to measure DNA methylation across approximately 3 kb of avpr1a in the retrosplenial cortex, a brain region implicated in male space use and sexual fidelity. We find dramatic variation in methylation across the avrp1a locus, with pronounced diversity near the exon-intron boundary and in a genetically variable putative enhancer within the intron. Among our wild voles, differences in cortical avpr1a expression correlate with DNA methylation in this putative enhancer, but not with the methylation status of the promoter. We also find an unusually high number of polymorphic CpG sites (polyCpGs) in this focal enhancer. One polyCpG within this enhancer (polyCpG 2170) may drive variation in expression either by disrupting transcription factor binding motifs or by changing local DNA methylation and chromatin silencing. Our results contradict some assumptions made within behavioural epigenetics, but are remarkably concordant with genome-wide studies of gene regulation.

History influences signal recognition: neural network models of túngara frogs.

Animals often attend to only a few of the cues provided by the complex displays of conspecifics. We suggest that these perceptual biases are influenced by mechanisms of signal recognition inherited from antecedent species. We tested this hypothesis by manipulating the evolutionary history of artificial neural networks, observing how the resulting networks respond to many novel stimuli and comparing these responses to the behaviour of females in phonotaxis experiments. Networks with different evolutionary histories proved equally capable of evolving to recognize the call of the túngara frog, Physalaemus pustulosus, but exhibited distinct responses to novel stimuli. History influenced the ability of networks to predict known responses of túngara frogs; network accuracy was determined by how closely the network history approximated the hypothesized history of the túngara frog. Our findings emphasize the influence of past selection pressures on current perceptual mechanisms, and demonstrate how neural network models can be used to address behavioural questions that are intractable through traditional methods.

Neural networks predict response biases of female túngara frogs.

Artificial neural networks have become useful tools for probing the origins of perceptual biases in the absence of explicit information on underlying neuronal substrates. Preceding studies have shown that neural networks selected to recognize or discriminate simple patterns may possess emergent biases toward pattern size of symmetry--preferences often exhibited by real females--and have investigated how these biases shape signal evolution. We asked whether simple neural networks could evolve to respond to an actual mate recognition signal, the call of the túngara frog, Physalaemus pustulosus. We found that not only were networks capable of recognizing the call of the túngara frog, but that they made remarkably accurate quantitative predictions about how well females generalized to many novel calls, and that these predictions were stable over several architectures. The data suggest that the degree to which P. pustulosus females respond to a call may often be an incidental by-product of a sensory system selected simply for species recognition.

Unilateral deficits induced in rats by MPP+ are markedly reduced by an N-terminal peptide fragment of dopamine-releasing protein.

Dopamine-releasing protein (DARP) is a novel factor involved in the function and development of catecholaminergic systems. To test whether a peptide synthesized from the N-terminus of DARP (DARP-36aa) ameliorates deficits in nigrostriatal dopamine, rats were unilaterally lesioned with MPP+, 1 day before administration of DARP-36aa began. Striatal delivery of 1 microgram DARPP-36aa daily for 10 days elevated striatal dopamine (P < 0.01) and reduced amphetamine-induced rotations (P < 0.05) relative to controls, both indicating the protection or restoration of dopaminergic function.

How evolutionary history shapes recognition mechanisms.

Evolutionary psychologists have emphasized the importance of natural selection in shaping cognitive functions, but historical contingency has not received direct study. This is crucial because in response to selection, complex traits tend to be fine-tuned or jury-rigged rather than totally reconstructed. We hypothesize that the neural and cognitive strategies an animal employs in signal recognition are influenced by the strategies used by its ancestors. The responses of female túngara frogs to ancestral calls and to calls of other closely related species are influenced by history. By training artificial neural networks with a series of calls that mimic the species' past history of call evolution or various control histories, we have shown that only networks that evolved through the mimetic history predict the response biases of túngara frogs.

Vestigial preference functions in neural networks and túngara frogs.

Although there is a growing interest in understanding how perceptual mechanisms influence behavioral evolution, few studies have addressed how perception itself is shaped by evolutionary forces. We used a combination of artificial neural network models and behavioral experiments to investigate how evolutionary history influenced the perceptual processes used in mate choice by female túngara frogs. We manipulated the evolutionary history of artificial neural network models and observed an emergent bias toward calls resembling known ancestral states. We then probed female túngara frogs for similar preferences, finding strong biases toward stimuli that resemble a call hypothesized for a recent ancestor. The data strongly suggest that female túngara frogs exhibit vestigial preferences for ancestral calls, and provide a general strategy for exploring the role of historical contingency in perceptual biases.

Regulation of male sexual behavior by progesterone receptor, sexual experience, and androgen.

Recent studies have demonstrated that physiological doses of progesterone may facilitate the androgen-dependent display of male sexual behavior in laboratory rats and three species of lizard. We used mice with a targeted disruption of the progesterone receptor to investigate whether such interactions exist in male mice and whether they may be modified by sexual experience. We found that naive intact male progesterone receptor knockout (PRKO) mice exhibit reduced mount frequencies compared to wild-type (WT) mice. Also unlike WT mice, sexually experienced PRKO males show profound losses in many measures of sexual behavior following castration. In a second experiment, we tested whether male mice heterozygous for a null mutation at the progesterone receptor locus were responsive to testosterone and progesterone treatment. We found that heterozygous males showed a reduced response to testosterone. The data are consistent with experiments indicating that the progesterone receptor is able to facilitate male-typical sex behaviors in other species and suggest novel mechanisms underlying the interaction of androgens and experience.