In this issue of Neuron, Miyasaka et al. demonstrate that the sequential transitions of male sexual behaviors are orchestrated by dual rhythms of dopamine and acetylcholine signaling generated in the ventral shell of the nucleus accumbens.

Male sexual behavior is innate and rewarding. Despite its centrality to reproduction, a molecularly specified neural circuit governing innate male sexual behavior and reward remains to be characterized. We have discovered a developmentally wired neural circuit necessary and sufficient for male mating. This circuit connects chemosensory input to BNSTpr neurons, which innervate POA neurons that project to centers regulating motor output and reward. Epistasis studies demonstrate that BNSTpr neurons are upstream of POA neurons, and BNSTpr-released substance P following mate recognition potentiates activation of POA neurons through Tacr1 to initiate mating. Experimental activation of POA neurons triggers mating, even in sexually satiated males, and it is rewarding, eliciting dopamine release and self-stimulation of these cells. Together, we have uncovered a neural circuit that governs the key aspects of innate male sexual behavior: motor displays, drive, and reward.

Social interactions require awareness and understanding of the behavior of others. Mirror neurons, cells representing an action by self and others, have been proposed to be integral to the cognitive substrates that enable such awareness and understanding. Mirror neurons of the primate neocortex represent skilled motor tasks, but it is unclear if they are critical for the actions they embody, enable social behaviors, or exist in non-cortical regions. We demonstrate that the activity of individual VMHvl neurons in the mouse hypothalamus represents aggression performed by self and others. We used a genetically encoded mirror-TRAP strategy to functionally interrogate these aggression-mirroring neurons. We find that their activity is essential for fighting and that forced activation of these cells triggers aggressive displays by mice, even toward their mirror image. Together, we have discovered a mirroring center in an evolutionarily ancient region that provides a subcortical cognitive substrate essential for a social behavior.

Sex hormones exert a profound influence on gendered behaviors. How individual sex hormone-responsive neuronal populations regulate diverse sex-typical behaviors is unclear. We performed orthogonal, genetically targeted sequencing of four estrogen receptor 1-expressing (Esr1) populations and identified 1,415 genes expressed differentially between sexes or estrous states. Unique subsets of these genes were distributed across all 137 transcriptomically defined Esr1 cell types, including estrous stage-specific ones, that comprise the four populations. We used differentially expressed genes labeling single Esr1 cell types as entry points to functionally characterize two such cell types, BNSTpr and VMHvl. We observed that these two cell types, but not the other Esr1 cell types in these populations, are essential for sex recognition in males and mating in females, respectively. Furthermore, VMHvl cell type projections are distinct from those of other VMHvl cell types. Together, projection and functional specialization of dimorphic cell types enables sex hormone-responsive populations to regulate diverse social behaviors.

Males as compared to females display increased impulsivity and inefficient inhibitory control and are more frequently diagnosed with disorders characterized by impulsivity. We previously demonstrated male rats make more impulsive action responses (i.e. premature responding) than females on the 5-choice serial reaction time task (5-CSRTT). Furthermore, pre-pubertal male rats make more impulsive choice responses (i.e. choosing an immediate small reward over a delayed larger reward) than females on a delayed-based reward T-maze task. The goal of the current work was to determine if gonadal hormones impact sex differences in impulsivity in adult rats. In an initial experiment, male and female rats underwent sham surgeries or were gonadectomized either pre-pubertally or during adulthood and tested on the 5-CSRTT in adulthood. Males displayed more impulsive action responses than females regardless of hormone status. In a second experiment, females received testosterone or vehicle injections on postnatal days 1 and 2. Males received vehicle injections. All rats were gonadectomized prior to puberty and tested on the 5-CSRTT in adulthood. Females treated neonatally with testosterone and control males made more impulsive action responses than control females. In another set of experiments, manipulation of gonadal hormones led to no differences in performance on the delayed-based reward T-maze task in males and females. Results indicate that no sex difference is apparent in impulsive choice on a delayed-base reward task in adult rats. They also reveal that adult sex differences on a task of impulsive action is mediated by organizational effects of gonadal hormones acting during the neonatal period and not impacted by hormones acting during puberty or adulthood.

Sexually naive animals have to distinguish between the sexes because they show species-typical interactions with males and females without meaningful prior experience. However, central neural pathways in naive mammals that recognize sex of other individuals remain poorly characterized. We examined the role of the principal component of the bed nucleus of stria terminalis (BNSTpr), a limbic center, in social interactions in mice. We find that activity of aromatase-expressing BNSTpr (AB) neurons appears to encode sex of other animals and subsequent displays of mating in sexually naive males. Silencing these neurons in males eliminates preference for female pheromones and abrogates mating success, whereas activating them even transiently promotes male-male mating. Surprisingly, female AB neurons do not appear to control sex recognition, mating, or maternal aggression. In summary, AB neurons represent sex of other animals and govern ensuing social behaviors in sexually naive males.

The unique hormonal, genetic and epigenetic environments of males and females during development and adulthood shape the neural circuitry of the brain. These differences in neural circuitry result in sex-typical displays of social behaviours such as mating and aggression. Like other neural circuits, those underlying sex-typical social behaviours weave through complex brain regions that control a variety of diverse behaviours. For this reason, the functional dissection of neural circuits underlying sex-typical social behaviours has proved to be difficult. However, molecularly discrete neuronal subpopulations can be identified in the heterogeneous brain regions that control sex-typical social behaviours. In addition, the actions of oestrogens and androgens produce sex differences in gene expression within these brain regions, thereby highlighting the neuronal subpopulations most likely to control sexually dimorphic social behaviours. These conditions permit the implementation of innovative genetic approaches that, in mammals, are most highly advanced in the laboratory mouse. Such approaches have greatly advanced our understanding of the functional significance of sexually dimorphic neural circuits in the brain. In this review, we discuss the neural circuitry of sex-typical social behaviours in mice while highlighting the genetic technical innovations that have advanced the field.

Impulsive actions and decisions often lead to undesirable outcomes. Lesion and neuroimaging studies have revealed that the orbital frontal cortex (OFC) and dorsal striatum (dSTR) play key roles in inhibitory control. It has been proposed that greater OFC input into the dSTR reflects enhanced top-down cognitive control and less impulsive responding. We previously reported a sex difference in inhibitory control, such that female rats make fewer impulsive errors than do male rats. The goal of the present study was to investigate differences in the OFC and dSTR of young adult male and female rats. In Experiment 1, we measured levels of two myelin-associated proteins, myelin basic protein (MBP) and myelin proteolipid protein (PLP), in the OFC and dSTR. Western blot data revealed that females had significantly higher levels of both MBP and PLP in the OFC but similar levels in the dSTR as compared to males. In Experiment 2, we infused the anterograde tracer, biotinylated dextran amine (BDA), into the OFC and measured the density of BDA in the dSTR. BDA was visualized using histochemistry followed by light microscopy imaging and densitometry analysis. Density of BDA in the dSTR was significantly greater in females as compared to males indicating that the projections from the OFC to dSTR may be greater in females as compared to males. Our results suggest a potential neuroanatomical sex difference that may contribute to the reported differences in inhibitory control levels of male and female rats.

The spontaneously hypertensive rat (SHR) is a commonly used and well-studied rodent model of attention deficit hyperactivity disorder (ADHD). Sex differences in the cognitive symptoms of ADHD are reported. However, the female SHR rat is much less studied than its male counterpart. The goal of the current study was to assess the validity of the SHR rodent model of ADHD by examining attentional performance, inhibitory control, and hyperactivity in both male and female SHR rats. Adult SHR and control Wistar-Kyoto rats were trained on the 5-choice serial reaction time task, a self-paced test of attention and inhibitory control. This task requires animals to identify the location of a brief light stimulus among five possible locations under several challenging conditions. Analyses of percent correct revealed that attentional performance in SHR females was not significantly different from control females, whereas attentional performance in SHR males was significantly different from control males. Analyses of the number of premature responses revealed that SHR rats made more inhibitory control errors than did control rats and that this decrease in inhibitory control was present in both SHR males and females. Analyses of activity in the open field revealed that SHR rats were more hyperactive than were control rats and that this increased hyperactivity was present in both SHR males and females. The current findings have implications for the study of sex differences in ADHD and for the use of SHR rats as a model of ADHD in females.

Neonatal testosterone, either acting directly or through its conversion to estradiol, can exert organizational effects on the brain and behavior. The goal of the current study was to examine sex differences and determine the role of neonatal testosterone on prefrontal cortex-dependent impulsive choice behavior in prepubertal rats. Male and female prepubertal rats were tested on the delay-based impulsive choice task. Impulsive choice was defined as choosing an immediate small food reward over a delayed large reward. In a first experiment to examine sex differences, males made significantly more impulsive choices than did females. In a second experiment to examine the organizational effects of testosterone, females treated with neonatal testosterone made significantly more impulsive choices than did control females and their performance was indistinguishable from that of control males. In a third experiment to determine if the effect of testosterone on performance is due to the actions of androgens or estrogens through its conversion to estradiol, males treated neonatally with the aromatase inhibitor formestane, which blocks the conversion of testosterone to estradiol, females treated neonatally with the non-aromatizable androgen dihydrotestosterone, and females treated neonatally with estradiol made significantly more impulsive choices than did control females and their performance was indistinguishable from that of control males. Results indicate that male pubertal rats display increased impulsive choice behavior as compared to females, that this sex difference results from organizing actions of testosterone during the neonatal period, and that this effect can result from both androgenic and estrogenic actions.

The goal of the current study was to explore sex differences in attentional processes in adult rats using a test of attention shown to be dependent upon the prefrontal cortex. Male and female adult Long-Evans rats were trained on the 5-choice serial reaction time task. This task requires rats to identify the location of a brief light stimulus among five possible locations. Performance was assessed under baseline conditions and under behavioral challenge conditions during which task difficulty was increased. Behavioral challenge conditions included shortening the stimulus duration, shortening the time before the onset of the stimulus, lengthening the time before the onset of the stimulus, and presenting a distracting noise. Analyses across baseline and all challenge conditions revealed that vigilance or sustained attention was more disrupted in female rats than it was in male rats, as measured by percent correct and number of omissions. Analyses also revealed that inhibitory control was more disrupted in male rats than it was in female rats, as measured by number of premature responses. These differences were most prominent when the onset of the stimulus was unpredictably lengthened. There were no differences in reward collection latency or correct response latency indicating no differences in motivation or sensory processes between the sexes. These results indicate that under challenging conditions adult female rats are more prone to make errors of vigilance than are adult male rats, and adult male rats are more prone to make errors of inhibitory control than are adult female rats.