Laura E. Been

Chemosensory and hormone information are relayed directly between the medial amygdala, posterior bed nucleus of the stria terminalis, and medial preoptic area in male Syrian hamsters.

In many rodent species, including Syrian hamsters, the expression of appropriate social behavior depends critically on the perception and identification of conspecific odors. The behavioral response to these odors is mediated by a network of steroid-sensitive ventral forebrain nuclei including the medial amygdala (Me), posterior bed nucleus of the stria terminalis (BNST), and medial preoptic area (MPOA). Although it is well-known that Me, BNST, and MPOA are densely interconnected and each uniquely modulates odor-guided social behaviors, the degree to which conspecific odor information and steroid hormone cues are directly relayed between these nuclei is unknown. To answer this question, we injected the retrograde tracer, cholera toxin B (CTB), into the BNST or MPOA of male subjects and identified whether retrogradely-labeled cells in Me and BNST 1) expressed immediate early genes (IEGs) following exposure to male and/or female odors or 2) expressed androgen receptor (AR). Although few retrogradely-labeled cells co-localized with IEGs, a higher percentage of BNST- and MPOA-projecting cells in the posterior Me (MeP) expressed IEGs in response to female odors than to male odors. The percentage of retrogradely-labeled cells that expressed IEGs did not, however, differ between and female and male odor-exposed groups in the anterior Me (MeA), posterointermediate BNST (BNSTpi), or posteromedial BNST (BNSTpm). Many retrogradely-labeled cells co-localized with AR, and a higher percentage of retrogradely-labeled MeP and BNSTpm cells expressed AR than retrogradely-labeled MeA and BNSTpi cells, respectively. Together, these data demonstrate that Me, BNST, and MPOA interact as a functional circuit to process sex-specific odor cues and hormone information in male Syrian hamsters.

The role of the medial preoptic area in appetitive and consummatory reproductive behaviors depends on sexual experience and odor volatility in male Syrian hamsters.

In Syrian hamsters (Mesocricetus auratus), the expression of reproductive behavior requires the perception and discrimination of sexual odors. The behavioral response to these odors is mediated by a network of ventral forebrain nuclei, including the medial preoptic area (MPOA). The role of MPOA in male copulatory behavior has been well-studied, but less is known about the role of MPOA in appetitive aspects of male reproductive behavior. Furthermore, many previous studies that examined the role of MPOA in reproductive behavior have used large lesions that damaged other nuclei near MPOA or fibers of passage within MPOA, making it difficult to attribute post-lesion deficits in reproductive behavior to MPOA specifically. Thus, the current study used discrete, excitotoxic lesions of MPOA to test the role of this nucleus in opposite-sex odor preference and copulatory behavior in both sexually-naïve and sexually-experienced males. Lesions of MPOA eliminated preference for volatile, opposite-sex odors in sexually-naïve, but not sexually-experienced, males. When males were allowed to contact the sexual odors, however, preference for female odors remained intact. Surprisingly, lesions of MPOA caused severe copulatory deficits only in sexually-naïve males, suggesting previous reports of copulatory deficits following MPOA lesions in sexually-experienced males were not due to damage to MPOA itself. Together, these results demonstrate that the role of MPOA in appetitive and consummatory aspects of reproductive behavior varies with the volatility of the sexual odors and the sexual experience of the male.

Lesions of the posterior bed nucleus of the stria terminalis eliminate opposite-sex odor preference and delay copulation in male Syrian hamsters: role of odor volatility and sexual experience.

In Syrian hamsters (Mesocricetus auratus), the expression of reproductive behavior requires the perception of social odors. The behavioral response to these odors is mediated by a network of ventral forebrain nuclei, including the posterior bed nucleus of the stria terminalis (pBNST). Previous studies have tested the role of the pBNST in reproductive behavior, but the use of large, fiber‐damaging lesions in these studies make it difficult to attribute post‐lesion deficits to the pBNST specifically. Thus, the current study used discrete, excitotoxic lesions of the pBNST to test the role of the pBNST in opposite‐sex odor preference and copulatory behavior in both sexually‐naive and sexually‐experienced males. Lesions of the pBNST decreased sexually‐naive males’ investigation of volatile female odors, resulting in an elimination of opposite‐sex odor preference. This elimination of preference was not due to a sensory deficit, as males with pBNST lesions were able to discriminate between odors. When, however, subjects were given sexual experience prior to pBNST lesions, their preference for volatile opposite‐sex odors remained intact post‐lesion. Similarly, when sexually‐naive or sexually‐experienced subjects were allowed to contact the social odors during the preference test, lesions of the pBNST decreased males’ investigation of female odors but did not eliminate preference for opposite‐sex odors, regardless of sexual experience. Finally, lesions of the pBNST delayed the copulatory sequence in sexually‐naive, but not sexually‐experienced, males such that they took longer to mount, intromit, ejaculate and display long intromissions. Together, these results demonstrate that the pBNST plays a unique and critical role in both appetitive and consummatory aspects of male reproductive behaviors.

Dissociated functional pathways for appetitive and consummatory reproductive behaviors in male Syrian hamsters

In many species, including Syrian hamsters, the generation of male reproductive behavior depends critically on the perception of female odor cues from conspecifics in the environment. The behavioral response to these odors is mediated by a network of steroid-sensitive ventral forebrain nuclei including the medial amygdala (MA), posterior bed nucleus of the stria terminalis (BNST) and medial preoptic area (MPOA). Previous studies have demonstrated that each of these three nuclei is required for appropriate sexual behavior and that MA preferentially sends female odor information directly to BNST and MPOA. It is unknown, however, how the functional connections between MA and BNST and/or MPOA are organized to generate different aspects of reproductive behavior. Therefore, the following experiments used the asymmetrical pathway lesion technique to test the role of the functional connections between MA and BNST and/or MPOA in odor preference and copulatory behaviors. Lesions that functionally disconnected MA from MPOA eliminated copulatory behavior but did not affect odor preference. In contrast, lesions that functionally disconnected MA from BNST eliminated preference for volatile female odors but did not affect preference for directly contacted odors or copulatory behavior. These results therefore demonstrate a double dissociation in the functional connections required for attraction to volatile sexual odors and copulation and, more broadly, suggest that appetitive and consummatory reproductive behaviors are mediated by distinct neural pathways.

ΔJunD overexpression in the nucleus accumbens prevents sexual reward in female Syrian hamsters

Motivated behaviors, including sexual experience, activate the mesolimbic dopamine system and produce long‐lasting molecular and structural changes in the nucleus accumbens. The transcription factor ΔFosB is hypothesized to partly mediate this experience‐dependent plasticity. Previous research in our laboratory has demonstrated that overexpressing ΔFosB in the nucleus accumbens of female Syrian hamsters augments the ability of sexual experience to cause the formation of a conditioned place preference. It is unknown, however, whether ΔFosB‐mediated transcription in the nucleus accumbens is required for the behavioral consequences of sexual reward. We therefore used an adeno‐associated virus to overexpress ΔJunD, a dominant negative binding partner of ΔFosB that decreases ΔFosB‐mediated transcription by competitively heterodimerizing with ΔFosB before binding at promotor regions on target genes, in the nucleus accumbens. We found that overexpression of ΔJunD prevented the formation of a conditioned place preference following repeated sexual experiences. These data, when coupled with our previous findings, suggest that ΔFosB is both necessary and sufficient for behavioral plasticity following sexual experience. Furthermore, these results contribute to an important and growing body of literature demonstrating the necessity of endogenous ΔFosB expression in the nucleus accumbens for adaptive responding to naturally rewarding stimuli.

The Neurobiology of Sexual Solicitation: Vaginal Marking in Female Syrian Hamsters (Mesocricetus auratus)

Vaginal marking is a reproductively-oriented scent marking response, in which female Syrian hamsters deposit vaginal secretions in response to odor cues from male conspecifics. Converging lines of evidence suggest that vaginal marking functions as a solicitational signal, or an advertisement of a female’s impending sexual receptivity. Although vaginal marking is commonly used as an assay of proceptivity, the neural control of vaginal marking remains largely unknown. In this chapter, we will review the existing literature on vaginal marking, synthesizing evidence from behavioral, endocrine, and neuroanatomical studies that indicate targets for the neural control of vaginal marking. Lastly, we will describe preliminary data from our laboratory that suggests a possible neural circuit for the descending control of vaginal marking in female Syrian hamsters.

X-ray kinematics analysis of vaginal scent marking in female Syrian hamsters (Mesocricetus auratus)

Vaginal marking is a stereotyped scent marking behavior in female Syrian hamsters used to attract male hamsters for mating. Although the modulation of vaginal marking by hormones and odors is well understood, the motor control of this proceptive reproductive behavior remains unknown. Therefore, we used X-ray videography to visualize individual bone movements during vaginal marking. Kinematic analyses revealed several consistent motor patterns of vaginal marking. Despite exhibiting a diversity of trial-to-trial non-marking behaviors (e.g. locomotor stepping), we found that lowering and raising the pelvis consistently corresponded with coordinated flexion and extension cycles of the hip, knee, and tail, suggesting that these movements are fundamental to vaginal marking behavior. Surprisingly, we observed only small changes in the angles of the pelvic and sacral regions, suggesting previous reports of pelvic rotation during vaginal marking may need to be reconsidered. From these kinematic data, we inferred that vaginal marking is primarily due to the actions of hip and knee extensor muscles of the trailing leg working against gravity to support the weight of the animal as it controls the descent of the pelvis to the ground. The cutaneous trunci muscle likely mediates the characteristic flexion of the tail. Interestingly, this tail movement occurred on the same time scale as the joint kinematics suggesting possible synergistic recruitment of these muscle groups. These data therefore provide new targets for future studies examining the peripheral control of female reproductive behaviors.

Metabotropic Glutamate Receptor and Fragile X Signaling in a Female Model of Escalated Aggression

BACKGROUND Escalated aggression is a behavioral sign of numerous psychiatric disorders characterized by a loss of control. The neurobiology underlying escalated aggression is unknown and is particularly understudied in females. Research in our laboratory demonstrated that repeated aggressive experience in female hamsters resulted in an escalated response to future aggressive encounters and an increase in dendritic spine density on nucleus accumbens (NAc) neurons. We hypothesized that the activation of group I metabotropic glutamate receptor signaling though the fragile X mental retardation protein (FMRP) pathway may underlie synaptic plasticity associated with aggression escalation. METHODS Female hamsters were given five daily aggression tests with or without prior treatment with the metabotropic glutamate receptor 5 (mGluR5) antagonist 2-methyl-6-(phenylethynyl)-pyridine. Following aggression testing, messenger RNA expression and protein levels were measured in the nucleus accumbens for postsynaptic density protein 95 (PSD-95) and SAP90/PSD-95-associated protein 3, as well as the levels of phosphorylated FMRP. RESULTS Experience-dependent escalation of aggression in female hamsters depends on activation of mGluR5 receptors. Furthermore, aggressive experience decreases phosphorylation of FMRP in the NAc, which is coupled to a long-term increase in the expression of the synaptic scaffolding proteins PSD-95 and SAP90/PSD-95-associated protein 3. Finally, the experience-dependent increase in PSD-95 is prevented by antagonism of the mGluR5 receptor. CONCLUSIONS Activation of the FMRP pathway by group I metabotropic glutamate receptors is involved in regulating synaptic plasticity following aggressive experience. The NAc is a novel target for preclinical studies of the treatment of escalated aggression, with the added benefit that emerging therapeutic approaches are likely to be effective in treating pathologic aggression in both female and male subjects.

Vesicular glutamate transporter 2 and tyrosine hydroxylase are not co-localized in Syrian hamster nucleus accumbens afferents

The nucleus accumbens (NAc) is an important brain region for motivation, reinforcement, and reward. Afferents to the NAc can be divided into two anatomically segregated neurochemical phenotypes: dopaminergic inputs, primarily from the midbrain ventral tegmental area (VTA) and glutamatergic inputs from several cortical and sub-cortical structures. A population of glutamatergic neurons exists within the VTA and evidence from rats and mice suggests that these VTA axons may co-release dopamine and glutamate into the NAc. Our laboratory has used sexual experience in Syrian hamsters as a model of experience-dependent plasticity within the NAc. Given that both dopamine and glutamate are involved in this plasticity, it is important to determine whether these neurotransmitters are co-expressed within the mesolimbic pathway of hamsters. We therefore used immunofluorescent staining to investigate the possible co-localization of tyrosine hydroxylase (TH), a dopaminergic marker, and vesicular glutamate transporter 2 (VGLUT2), a glutamatergic marker, within the mesolimbic pathway. PCR analyses identified VGLUT2 gene expression in the VTA. No co-localization of TH and VGLUT2 protein was detected in NAc fibers, nor was there a difference in immunolabeling between males and females. Further studies are needed to resolve this absence of anatomical co-localization of TH and VGLUT2 in hamster striatal afferents with reports of functional co-release in other rodents.