Male rat sexual behavior requires aromatization of testosterone (T) to estradiol (E2) in the medial preoptic area (MPO) where estrogen receptors (ER) exist in two isoforms, ERα and ERβ. We hypothesized that E2 acts through estrogen receptor α (ERα) in the MPO to promote male mating behavior. Four groups of male rats were castrated, administered DHT s.c. and bilateral MPO implants delivering either: cholesterol, E2, propyl pyrazole triol (PPT, ERα agonist), diarylpropionitrile (DPN, ER β agonist), or 1-methyl-4-phenyl pyridinium (MPP, ERα antagonist). Additional gonadally intact males received bilateral MPO DPN implants. PPT maintained sexual behavior equally as well as E2, whereas mating was not maintained by cholesterol or DPN MPO implants. Exogenous T did not reinstate mating in animals that received MPP MPO implants. These findings indicate that, in the MPO, ERα is necessary and sufficient to promote copulatory behavior in male rats and ERβ is not sufficient for mating.

Thesis advisor: Michael Numan / Postpartum female rats immediately respond to biological or foster offspring with the display of maternal behavior. In contrast, females that are hysterectomized and ovariectomized on day 15 of pregnancy (15HO) and presented with pups 48 hours later show maternal behavior after 2-3 days of pup exposure. The natural onset of maternal behavior in postpartum females is mediated, in part, by the rise in estradiol just prior to birth. When 15HO rats are administered estradiol benzoate (EB) at the time of HO surgery, 48 hours prior to pup presentation, they show an almost immediate onset of maternal behavior. Presumably, EB administration functions to prime neural circuits which regulate maternal behavior such that these circuits respond to pup presentation with increased maternal responsiveness. Two important neural regions which have been shown to interact in order to promote responsiveness toward infant stimuli are the medial preoptic area (MPOA) and the nucleus accumbens (NA). The following series of studies were undertaken to examine how dopamine (DA) activity within these two important neural sites substitutes for the facilitatory effects of chronic (48 hours) EB stimulation of maternal behavior in 15HO rats. Study 1 investigates whether, in the absence of EB treatment, microinjection of dopamine receptor agonists into either NA or MPOA at the time of pup presentation stimulate maternal behavior in 15HO rats. Study 2 examines the underlying mechanism by which DA receptor stimulation of NA promotes the onset of maternal behavior in 15HO rats. Finally, Study 3 examines the relationship between DA receptor stimulation and estradiol stimulation in the facilitation of maternal responsiveness in 15HO rats. / Thesis (PhD) — Boston College, 2009. / Submitted to: Boston College. Graduate School of Arts and Sciences. / Discipline: Psychology.

Maternal behaviours are those that increase offspring survival. Estrogens affect maternal behaviour by activating Estrogen Receptors (ER) in the brain. Maternal brain plasticity was explored by characterizing the effects of exposure to a mixture of environmental pollutants on number of ERs. Following exposure to the toxicants during pregnancy and lactation, brains of female rats were collected, sectioned at 30 μm and immunohistochemistry for ERα performed. Immuno-positive cells in the mPOA, VTA and NAc were counted. A two way ANOVA revealed no main effect of Treatment on the number of immunopositive cells for all three brain regions. However, a significant difference between the High and Low Doses with the high dose reducing the number of ERα+ cells in the mPOA and VTA. Our work showcases the importance of studying the effects of multiple chemical co-exposures on the mother's brain, as maternal brain changes impact maternal behaviour consequently affecting offspring neurodevelopment.

Male rat mating is a suite of individual behaviors mediated by the actions of two metabolites of testosterone (T), dihydrotestosterone (DHT) and estradiol (E2), on the brain. Individually, neither metabolite fully maintains or restores mating in castrated males, but both combined are as effective as T. Two hormone-responsive areas of the brain, the medial preoptic area (MPO) and the medial amygdala (MEA), are crucial for mating. These studies ask: by what mechanism(s) does E2 act in the MPO and MEA? We blocked the conversion of T to E2 in the MEA of intact male rats and sexual behavior was not maintained. We then infused antisense oligodeoxynucleotides (ODNs) to estrogen receptor-alpha (ER-á) mRNA bilaterally to the MPO or the MEA of intact male rats to block ER-á expression. ODN infusion of the MPO attenuated mating but infusion of the MEA had no effect. These results suggest that ER-á is the behaviorally relevant estrogen receptor (ER) in the MPO but not in the MEA. ER was originally described in the cytoplasm and nucleus of cells. Recently plasma membrane associated ERs (mER) have been reported. We conjugated E2 to Bovine Serum Albumin (BSA-E2), a large protein that will not penetrate the plasma membrane, thus restricting the action of E2 to mER, and chronically delivered it to the MPO and MEA. BSA-E2 maintained mating if put in the MPO, but not in the MEA, suggesting a surface action of E2 is sufficient in the MPO. The MPO and MEA are reciprocally connected and probably constitute elements of a larger, steroid-responsive neural network that mediates male mating behavior. To begin to describe this purported circuit, we injected Pseudorabies virus (PRV) into the prostate gland and dually labeled PRV-immunoreactive cells for ER or androgen receptors. We found dual labeling in a forebrain diencephalic circuit that includes the MPO, the medial preoptic nucleus, bed nucleus of stria terminalis, the zona incerta, the periaqueductal gray and other areas that presumably mediate both autonomic and motor aspects of male mating. Together, the results of these studies begin to elucidate locations and mechanisms of E2 mediation of male sexual behavior.

Male rat copulation is mediated by estrogen-sensitive neurons in the medial preoptic area (MPO) and medial amygdala (MEA); however, the mechanisms through which estradiol (E2) acts are not fully understood. We hypothesized that E2 acts through estrogen receptor α (ERα) in the MPO and MEA to promote male mating behavior. Antisense oligodeoxyneucleotides (AS-ODN) complementary to ERα mRNA were bilaterally infused via minipumps into either brain area to block the synthesis of ERα, which we predicted would reduce mating. Western blot analysis and immunocytochemistry revealed a knockdown of ERα in each brain region; however, compared to saline controls, males receiving AS-ODN to the MPO showed significant reductions in all components of mating, whereas males receiving AS-ODN to the MEA continued to mate normally. These results suggest that E2 acts differently in these brain regions to express sexual behavior and that ERα in the MPO, but not in the MEA, promotes mating.

In many species, including Syrian hamsters, male reproductive behavior depends on the perception of odor cues from conspecifics in the environment. Volatile odor cues are processed primarily by the main olfactory system, whereas non-volatile cues are processed primarily by the accessory olfactory system. Together, these two chemosensory systems mediate appetitive reproductive behaviors, such as attraction to female odors, and consummatory reproductive behaviors, such as copulation, in male Syrian hamsters. Main and accessory olfactory information are first integrated in the medial amygdala (MA), a limbic nucleus that is critical for the expression of reproductive behaviors. MA is densely interconnected with other ventral forebrain nuclei that receive chemosensory information and are sensitive to steroid hormones. Specifically, several lines of evidence suggest that MA may generate behavioral responses to socio-sexual odors via functional connections with the posterior bed nucleus of the stria terminalis (BNST) and medial preoptic area (MPOA). It is unknown, however, how these three nuclei act as functional circuit to adaptively regulate appetitive and consummatory reproductive behaviors. Therefore, the overarching goal of this dissertation was to determine how BNST and MPOA function, both uniquely and as a circuit with MA, to generate attraction to female odors and copulatory behaviors in male Syrian hamsters. We found that BNST is required for attraction to female odors, but not for copulation, in sexually-naïve males. In contrast, MPOA is required for both attraction to female odors and for copulation in sexually-naïve males. Surprisingly, prior sexual experience mitigated the requirement of BNST and MPOA for these behaviors. Next, we found that MA preferentially transmits female odor information to BNST and to MPOA, whereas BNST relays female and male odor information equivalently to MPOA. Finally, we found that the functional connections between MA and BNST are required for attraction to female odors but not for copulation, whereas the functional connections between MA and MPOA are required for copulation but not for attraction to female odors. Ultimately, these data may uncover a fundamental mechanism by which this ventral forebrain circuit regulates appetitive and consummatory reproductive behaviors across many species and modalities.

The salience of natural or drug-associated reward is mediated by phasic dopamine (DA) release in the nucleus accumbens (NAc) arising from DAergic cells in the ventral tegmental area (VTA). Circulating sex steroid hormones can modulate reward associated with drugs of abuse; yet, it still remains unclear which brain regions are responsible for this modulation. The medial preoptic area (mPOA) is a hypothalamic brain area involved in the expression of naturally rewarding behaviors as well as the regulation and reception of circulating sex steroid hormones in female rats. Considering its role in regulating naturally rewarding behaviors, its well-established anatomical connectivity with the VTA, and its responsiveness to circulating sex steroid hormones, the mPOA is an ideal neural node through which hormones could modulate the rewarding facets of drugs of abuse. Here I show that preoptotegmental efferents to the VTA are primarily GABAergic, that they appose putative DAergic cell bodies in the VTA that project to the NAc, and that they are capable of responding to sex steroid hormones and changes in DA release. Furthermore, cocaine influences neural activity in mPOA efferents that project to the VTA. Removal of the mPOA also enhanced cocaine-induced locomotion, Fos-immunoreactivity in the mesolimbic reward system, DA release in the NAc, and augmented conditioned place preference. Together these findings suggest that the mPOA modulates the release of DA in the mesolimbic reward circuitry via inhibitory connections with DA neurons residing in the VTA, and sex steroid hormones, in turn, may act in the mPOA to modulate response to cocaine. / text

Mice display robust and stereotyped behaviors towards pups: virgin males typically attack pups, while virgin females and sexually experienced males display parental care. I show here that virgin males that are genetically impaired in vomeronasal sensing do not attack pups and are parental, suggesting a key role of the vomeronasal system in controlling male infanticide. In addition, we have identified putative vomeronasal receptors (or receptor groups) for the detection of pup odors, thus uncovering new tools for the molecular and genetic dissection of male infanticide. Further, we have uncovered galanin-expressing neurons in the medial preoptic area (MPOA) as key regulators of male and female parental behavior. Genetic ablation of MPOA galanin- neurons results in dramatic impairment of parental responses in both virgin females and sexually experienced males. In addition, optogenetic activation of these cells in virgin males suppresses infanticide and induces pup grooming. Thus, MPOA galanin-expressing neurons emerge as an essential node of regulation of innate behavior in the hypothalamus that orchestrates male and female parenting while opposing vomeronasal circuits underlying infanticide. Our results provide an entry point for the genetic and circuit-level dissection of mouse parental behavior and its modulation by social experience.

ABSTRACT Evidence from knockout studies in male mice and from experiments in male rats,in which expression of the estrogen receptor alpha (ERα) gene was inhibited in the medial preoptic area (MPO), suggests that ERα is important in the control of male rat mating behavior. Therefore, in this experiment, we tested the hypothesis that activation of ERα in the MPO is sufficient to maintain mating behavior in castrated male rats receiving subcutaneously (s.c.) dihydrotestosterone (DHT), a non-aromatizable androgen. Accordingly, castrated rats treated with DHT s.c. received MPO implants of either: (i) propyl-pyrazole-triol (PPT) (Stauffer, et al 2000; Katzenellenbogen, et al 2000), a selective ERα agonist, (ii) E2 (positive controls) or (iii) cholesterol (negative controls)and sexual behavior was monitored. PPT was as effective as E2 at maintaining mating behavior suggesting that, in the MPO, ERα is sufficient to mediate responses to E2 that underlie male rat mating behavior.

In the present work we evaluated the degree of sexual dimorphism in two cell groups of the medial preoptic-anterior hypothalamus (MPOA-AH) in monogamous and polygamous voles. Quantitative determinations were made of volume, cell number, and cell density for the anteroventral-periventriular nucleus (AVPV) and the sexually dimorphic nucleus of the preoptic area (SDN-POA). Polygamous montane voles (Microtus montanus) had a greater degree of sexual dimorphism in both cell groups than did monogamous prairie voles (M. ochrogaster). Most notable was the complete absence of the AVPV in male montane voles; male montane voles also had a significantly larger SDN-POA volume than did females. The only sexual dimorphism in prairie voles was a greater cell density in the female AVPV. In addition, prairie voles had larger relative brain size than did montane voles. Comparative behavioral studies have revealed a correlation between the degree of sexual dimorphism in external morphology and mating system, i.e., polygamous species display greater levels of dimorphism than do monogamous species. The present results indicate that the effects of sexual selection can also be seen in those brain regions, like the hypothalamus, that underlie social and reproductive behavior. Moreover, these results support the hypothesis that neuroanatomic dimorphisms in the MPOA-AH may be related to sex differences in behavior.