Role of aromatase in distinct brain nuclei of the social behaviour network in the expression of sexual behaviour in male Japanese quail.

In male Japanese quail, brain aromatase is crucial for the hormonal activation of sexual behaviour, but the sites producing neuro-oestrogens that are critical for these behaviours have not been completely identified. This study examined the function of aromatase expressed in several nuclei of the social behaviour network on a measure of sexual motivation known as the frequency of rhythmic cloacal sphincter movements (RCSM) and on copulatory behaviour. Sexually experienced castrated males chronically treated with testosterone were stereotaxically implanted with the aromatase inhibitor vorozole (VOR), or cholesterol as control, and tested for sexual behaviour. In experiment 1, males were implanted in the medial preoptic nucleus (POM) with VOR, a manipulation known to reduce the expression of copulatory behaviour. This experiment served as positive control, but also showed that VOR implanted in the dorsomedial or lateral portions of the POM similarly inhibits male copulatory behaviour compared to control implants. In experiments 2 to 4, males received stereotaxic implants of VOR in the periaqueductal gray (PAG), the nucleus taeniae of the amygdala (TnA) and the ventromedial nucleus of the hypothalamus (VMN), respectively. Sexual behaviour was affected only in individuals where VOR was implanted in the PAG: these males displayed significantly lower frequencies of cloacal contact movements, the last step of the copulatory sequence. Inhibition of aromatase in the TnA and VMN did not alter copulatory ability. Overall, RCSM frequency remained unaffected by VOR regardless of implantation site. Together, these results suggest that neuro-oestrogens produced in the POM contribute the most to the control of male copulatory behaviour, while aromatase expressed in the PAG might also participate to premotor aspects of male copulatory behaviour.

Occupational Preferences and Recalled Childhood Sex-Atypical Behavior among Istmo Zapotec Men, Women, and Muxes.

Research has found that both cisgender and transgender androphilic males (i.e., males sexually attracted to and aroused by other adult males) have female-typical occupational preferences when compared with gynephilic males (i.e., males sexually attracted to and aroused by adult females). Moreover, whereas cisgender androphilic males' occupational preferences tend to be intermediate between those of gynephilic men and androphilic women, transgender androphilic males tend to have occupational preferences that are more similar to androphilic women. No study has directly compared both types of androphilic males within the same culture. The present study investigated occupational preference and its relation to childhood sex-atypical behavior (CSAB), among gynephilic men (n = 208), androphilic women (n = 138), and cisgender (n = 132) and transgender (n = 129) androphilic males from the Istmo region of Oaxaca, Mexico, where androphilic males are recognized as third gender, muxes. The study found large sex differences in occupational preferences (d = 2.80). Furthermore, both cisgender muxe nguiiu (d = 2.36) and transgender muxe gunaa (d = 3.44) reported having more sex-atypical occupational preferences compared with men. However, muxe gunaa reported higher female-typical occupational preferences than women (d = 0.59) and muxe nguiiu (d = 0.57), whereas muxe nguiiu and women did not differ (d = 0.08). These findings are consistent with the conclusion that sex-atypical occupational preferences are a cross-culturally universal aspect of male androphilia. Finally, CSAB was associated with sex-atypical occupational preferences among all participants. These findings suggest that a developmental continuity exists between childhood and adulthood sex-atypicality.

Effect of chronic intracerebroventricular administration of an aromatase inhibitor on the expression of socio-sexual behaviors in male Japanese quail.

Aromatase converts androgens into estrogens in the brain of vertebrates including humans. This enzyme is also expressed in other tissues where its action may result in negative effects on human health (e.g., promotion of tumor growth). To prevent these effects, aromatase inhibitors were developed and are currently used to block human estrogen-dependent tumors. In vertebrates including quail, aromatase is expressed in a highly conserved set of interconnected brain nuclei known as the social behavior network. This network is directly implicated in the expression of a large range of social behaviors. The primary goal of this study was to characterize in Japanese quail the potential impact of brain aromatase on sexual behavior, aggressiveness and social motivation (i.e., tendency to approach and stay close to conspecifics). An additional goal was to test the feasibility and effectiveness of long-term delivery of an aromatase inhibitor directly into the third ventricle via Alzet™ osmotic minipumps using male sexual behavior as the aromatase dependent measure. We demonstrate that this mode of administration results in the strongest inhibition of both copulatory behavior and sexual motivation ever observed in this species, while other social behaviors were variably affected. Sexual motivation and the tendency to approach a group of conspecifics including females clearly seem to depend on brain aromatase, but the effects of central estrogen production on aggressive behavior and on the motivation to approach males remain less clear.

Key role of estrogen receptor ß in the organization of brain and behavior of the Japanese quail.

Estrogens play a key role in the sexual differentiation of the brain and behavior. While early estrogen actions exert masculinizing effects on the brain of male rodents, a diametrically opposite effect is observed in birds where estrogens demasculinize the brain of females. Yet, the two vertebrate classes express similar sex differences in the brain and behavior. Although ERα is thought to play a major role in these processes in rodents, the role of ERß is still controversial. In birds, the identity of the estrogen receptor(s) underlying the demasculinization of the female brain remains unclear. The aim of the present study was thus to determine in Japanese quail the effects of specific agonists of ERα (propylpyrazole triol, PPT) and ERß (diarylpropionitrile, DPN) administered at the beginning of the sensitive period (embryonic day 7, E7) on the sexual differentiation of male sexual behavior and on the density of vasotocin-immunoreactive (VT-ir) fibers, a known marker of the organizational action of estrogens on the quail brain. We demonstrate that estradiol benzoate and the ERß agonist (DPN) demasculinize male sexual behavior and decrease the density of VT-ir fibers in the medial preoptic nucleus and the bed nucleus of the stria terminalis, while PPT has no effect on these measures. These results clearly indicate that ERß, but not ERα, is involved in the estrogen-induced sexual differentiation of brain and sexual behavior in quail.

A Retrospective Study of Childhood Sex-Typed Behavior in Istmo Zapotec Men, Women, and Muxes.

Previous research has consistently demonstrated that both transgender and cisgender androphilic males (i.e., males attracted to adult males) display and recall higher levels of childhood female-typical behavior (CFTB) and lower levels of childhood male-typical behavior (CMTB) compared to gynephilic males (i.e., males attracted to adult females). In adulthood, the recalled CFTB and CMTB scores of cisgender androphilic males tend to be intermediate to those of opposite-sex-attracted men and women, whereas transgender androphilic males tend to score similar to women. These studies have been mostly conducted in Euro-American cultures. We examined recalled childhood sex-typed behavior (CSTB) among the Istmo Zapotec-a pre-Colombian culture in the Istmo region of Oaxaca, Mexico, where cisgender and transgender androphilic males are recognized as a third gender, known locally as muxes. The present study sought to determine whether Istmo Zapotec men (n = 180), cisgender muxe nguiiu (n = 63), transgender muxe gunaa (n = 120), and women (n = 138) differ with respect to recalled CFTB and CMTB. Our results indicate that men recalled significantly less CFTB and more CMTB than women. Cisgender muxes scored in between men and women. Transgender muxes scored similar to women. These findings provide further evidence that childhood sex-atypical behavior is a cross-culturally universal and normative developmental aspect of male androphilia, regardless of whether it manifests in the cisgender or transgender form. This is the first study to present quantitative data comparing the recalled CSTB of cisgender and transgender androphilic males from within the same non-Euro-American culture.

Role for the membrane estrogen receptor alpha in the sexual differentiation of the brain.

Estrogens exert pleiotropic effects on multiple physiological and behavioral responses. Male and female sexual behavior in rodents constitutes some of the best-characterized responses activated by estrogens in adulthood and largely depend on ERα. Evidence exists that nucleus- and membrane-initiated estrogen signaling cooperate to orchestrate the activation of these behaviors both in short- and long-term. However, questions remain regarding the mechanism(s) and receptor(s) involved in the early brain programming during development to organize the circuits underlying sexually differentiated responses. Taking advantage of a mouse model harboring a mutation of the ERα palmitoylation site, which prevents membrane ERα signaling (mERα; ERα-C451A), this study investigated the role of mERα on the expression of male and female sexual behavior and neuronal populations that differ between sexes. The results revealed no genotype effect on the expression of female sexual behavior, while male sexual behavior was significantly reduced, but not abolished, in males homozygous for the mutation. Similarly, the number of kisspeptin- (Kp-ir) and calbindin-immunoreactive (Cb-ir) neurons in the anteroventral periventricular nucleus (AVPv) and the sexually dimorphic nucleus of the preoptic area (SDN-POA), respectively, were not different between genotypes in females. In contrast, homozygous males showed increased numbers of Kp-ir and decreased numbers of Cb-ir neurons compared to wild-types, thus leading to an intermediate phenotype between females and wild-type males. Importantly, females neonatally treated with estrogens exhibited the same neurochemical phenotype as their corresponding genotype among males. Together, these data provide evidence that mERα is involved in the perinatal programming of the male brain.

Effects of a novel partner and sexual satiety on the expression of male sexual behavior and brain aromatase activity in quail.

This study was designed to determine whether changes in sexual motivation acutely regulate brain estrogen synthesis by aromatase. Five experiments (Exp.1-5) were first conducted to determine the effect of recent mating and of the presentation of a new female (Coolidge effect) on sexual motivation. Exp.1-2 showed that 10 min or overnight access to copulation decreases measures of male sexual motivation when male subjects were visually exposed to the female they had copulated with and this effect is not counteracted by the view of a new female. Exp.3 showed that sexual motivation is revived by the view of a new female in previously unmated males only allowed to see another female for 10 min. After mating for 10 min (Exp.4) or overnight (Exp.5) with a female, males showed a decrease in copulatory behavior that was not reversed by access to a new female. Exp.6 and 7 confirmed that overnight copulation (Exp.6) and view of a novel female (Exp.7) respectively decreases and increases sexual behavior and motivation. Yet, these manipulations did not affect brain aromatase activity except in the tuberal hypothalamus. Together these data confirm that copulation or prolonged view of a female decrease sexual motivation but a reactivation of sexual motivation by a new female can only be obtained if males had only seen another female but not copulated with her, which is different in some degree from the Coolidge effect described in rodents. Moreover changes in brain aromatase do not simply reflect changes in motivation and more complex mechanisms must be considered.

Familial patterning and prevalence of male androphilia among Istmo Zapotec men and muxes.

Male androphilia (i.e., male sexual attraction to other adult males) is known to cluster within families. Some studies demonstrate that male androphilia clusters in both the paternal and maternal familial lines, whereas other studies demonstrated that it clusters only in the latter. Most of these studies were conducted in Euro-American populations where fertility is low and the sexual orientation of male relatives can sometimes be difficult to ascertain. These two factors can potentially confound the results of such studies. To address these limitations, we examined the familial patterning of male androphilia among the Istmo Zapotec of Oaxaca, Mexico--a high fertility, non-Euro-American population where androphilic males are known locally as muxes, a third gender category. The Istmo Zapotec recognize two types of muxes--muxe gunaa and muxe nguiiu--who typify the transgender and cisgender forms of male androphilia, respectively. We compared the familial patterning of male androphilia between muxe gunaa and muxe nguiiu, as well as between gynephilic men and muxes (both cisgender and transgender forms combined). Istmo Zapotec muxe gunaa and muxe nguiiu exhibit similar familial patterning of male androphilia. Overall, muxes were characterized by significantly more muxe relatives than gynephilic men. This familial patterning was equivalent in both the paternal and maternal lines of muxes. The population prevalence rate of male androphilia was estimated to fall between 3.37-6.02% in the Istmo Zapotec. This is the first study that has compared cisgender and transgender androphilic males from the same high fertility population and demonstrated that the two do not differ with respect to the familial patterning of male androphilia.

Recalled Separation Anxiety in Childhood in Istmo Zapotec Men, Women, and Muxes.

The Istmo Zapotec are a pre-Columbian cultural group indigenous to the Istmo region of Oaxaca, Mexico. Istmo Zapotec recognize three genders: men, women, and muxes. Like Istmo Zapotec men, muxes are biological males. However, unlike Istmo Zapotec men, most muxes are exclusively androphilic (i.e., sexually attracted to adult males), relatively feminine, and routinely adopt the receptive role during anal intercourse. Furthermore, the Istmo Zapotec recognize two types of muxes: muxe gunaa, who resemble the transgender androphilic males that are common in many non-Western cultures, and muxe nguiiu, who resemble the cisgender androphilic males ("gay" men) common in Western cultures. Retrospective research conducted in Canada and Samoa demonstrates that cisgender and transgender androphilic males recall elevated indicators of childhood separation anxiety (i.e., feelings of distress related to separation from major attachment figures) when compared to males who are gynephilic (i.e., sexually attracted to adult females). The present study compared recalled indicators of childhood separation anxiety among Istmo Zapotec men, women, muxe gunaa, and muxe nguiiu (N = 454). Men recalled significantly lower levels of childhood separation anxiety compared to all other groups (all p < .042). No additional group differences were found. Our results are consistent with previous research conducted in Canada and Samoa, suggesting that elevated childhood separation anxiety is a developmental correlate of male androphilia that is cross-culturally universal. This research is also consistent with the conclusion that cisgender and transgender male androphiles share a common biological and developmental foundation despite being different in appearance.