Recalled Childhood Separation Anxiety in Relation to Sex, Sexual Orientation, and Gender Identity in Thailand.

Same-sex sexual attraction is an evolutionary paradox. It is influenced by genes but also associated with reduced reproduction. An altruistic disposition toward kin could increase relatives' reproduction, thus addressing this paradox. Developmentally, such a disposition may manifest as anxiety during separation from kin. This hypothesis has been supported among androphilic (i.e. sexually attracted to males) males, but few similar studies of gynephilic (i.e. sexually attracted to females) females exist. We examined recalled childhood separation anxiety in Thailand and employed the largest and most sexually and gender-diverse sample to date (N = 1403). Among heterosexuals, men recalled less concern for parents' and siblings' well-being during separation than women. Androphilic males evidenced elevated concern for parents' and siblings' well-being during separation, and transfeminine androphilic males known as sao praphet song reported heightened anxiety when separated from parents. Among females, transmasculine gynephilic females known as toms recalled less concern about parents' well-being during separation than feminine females who are attracted to toms and known as dees. Bisexual women recalled less anxiety when separated from parents than dees and lesbian and heterosexual women. These findings suggest a disposition toward heightened kin-directed altruism is evident in childhood among androphilic males, but not gynephilic females.

Carving the Biodevelopment of Same-Sex Sexual Orientation at Its Joints.

Sexual orientation is a core aspect of human experience and understanding its development is fundamental to psychology as a scientific discipline. Biological perspectives have played an important role in uncovering the processes that contribute to sexual orientation development. Research in this field has relied on a variety of populations, including community, clinical, and cross-cultural samples, and has commonly focused on female gynephilia (i.e., female sexual attraction to adult females) and male androphilia (i.e., male sexual attraction to adult males). Genetic, hormonal, and immunological processes all appear to influence sexual orientation. Consistent with biological perspectives, there are sexual orientation differences in brain development and evidence indicates that similar biological influences apply across cultures. An outstanding question in the field is whether the hypothesized biological influences are all part of the same process or represent different developmental pathways leading to same-sex sexual orientation. Some studies indicate that same-sex sexually oriented people can be divided into subgroups who likely experienced different biological influences. Consideration of gender expression in addition to sexual orientation might help delineate such subgroups. Thus, future research on the possible existence of such subgroups could prove to be valuable for uncovering the biological development of sexual orientation. Recommendations for such future research are discussed.

Recalled Childhood Separation Anxiety Differs by Anal Sex Role among Gay Men.

Male androphilia (i.e., male sexual attraction to adult males) is considered an evolutionary paradox because it is partially influenced by genes and associated with decreased reproduction. Traits associated with attachment to genetic relatives (i.e., kin) could prompt increased kin-directed altruism, thereby offsetting decreased reproduction by helping kin reproduce. These traits include childhood separation anxiety and adulthood neuroticism, which have been associated with feminine gender expression. In prior research, gay men with a receptive (Bottom or Versatile) anal sex role (ASR) reported greater childhood gender nonconformity (GNC) than those with an insertive (Top) ASR. We examined whether ASR groups also differed on recalled childhood separation anxiety and adulthood neuroticism. The Separation Anxiety Scale-Revised and Big-Five Personality Inventory - short form were completed by 350 gay and 146 heterosexual men. For neuroticism, ASR preference groups differed from heterosexual men but not from one another. Gay men who preferred a Bottom or Versatile ASR reported higher recalled childhood separation anxiety than Tops and heterosexual men. Recalled childhood GNC mediated ASR group differences with heterosexual men on childhood separation anxiety. These results indicate that subgroups of gay men delineated by ASR differ on an evolutionarily relevant developmental trait, childhood separation anxiety.

Overexpression of Androgen Receptors Masculinizes 2D:4D Digit Ratios in Mice.

Studying the role of the prenatal endocrine environment in humans is challenging due to the ethical and practical considerations of measuring hormone levels of the developing fetus. Because it has been difficult to ascertain whether prenatal androgens contribute to the brain and behavior in humans as it does in non-human species, retrospective markers of prenatal androgens, such as the second-to-fourth finger digit ratio (2D:4D), are of interest to the studying of human behavioral endocrinology. To assess the validity of such markers, laboratory animals have been studied. Some strains of mice have been reported to show a sex difference in 2D:4D, and pharmacological and genetic manipulation of the androgen and estrogen receptors (AR and ER) has implicated a role for prenatal androgens in mediating this sex difference, although there have been conflicting reports. Here, we compared mice with global AR overexpression to mice with wildtype (WT) littermates and mice with neural-specific AR overexpression. We found a sex difference in the right hind paw, such that males had larger digit ratios than females. Regardless of sex, mice with global AR overexpression showed an increase in the right hind 2D:4D ratio compared with both WT and neural-specific AR overexpression mice. These results support a role for non-neural AR in the development of 2D:4D and suggest that increased sensitivity to androgens via increased AR is sufficient to increase the masculinization of digit ratios. Future directions for confirming the validity of 2D:4D as a marker for prenatal androgen exposure are discussed.

Mental rotation task performance in relation to sexual and gender diversity in Thailand.

Neurohormonal theory argues that organizational effects of hormone exposure influence sexual orientation and gender identity, as well as sex differences in visuospatial cognition. This study examined mental rotation task (MRT) performance in a diverse Thai sample (N = 980). Thai culture has several third genders: individuals assigned male at birth (AMAB) who are feminine and attracted to cis men (i.e., sao praphet song); individuals assigned female at birth (AFAB) who are masculine and attracted to feminine individuals (i.e., toms); AFAB individuals who are feminine and attracted to toms (i.e., dees); and sexual orientation categories similar to Western culture (e.g., gay, lesbian, bi). On the MRT, straight cis men outperformed straight cis women. Results were consistent with organizational effects among AMAB individuals, with straight cis men outperforming gay cis men and sao praphet song. Among AFAB individuals, however, only bi and lesbian cis women outperformed dees. Overall, support for neurohormonal theory was limited among AFAB individuals, but MRT performance among AMAB individuals was consistent with organizational effects. This study informs our understanding of visuospatial sex/gender differences and the applicability of neurohormonal theory across cultures.

An anthropometric study of sexual orientation and gender identity in Thailand.

The biodevelopment of psychological sex differentiation is putatively reflected in several anthropometrics. We examined eight anthropometrics in 1404 Thai participants varying in sex, sexual orientation, and gender identity/expression: heterosexual men and women, gay men, lesbian women, bisexual women, sao praphet song (transgender birth-assigned males), toms (transgender birth-assigned females), and dees (birth-assigned females attracted to toms). Exploratory factor analyses indicated the biomarkers should be analyzed independently. Using regressions, in birth-assigned males, less male-typical second-to-fourth digit ratios in the left hand were associated with sexual orientation towards men regardless of gender identity/expression, whereas shorter height and long-bone growth in the arms and legs were more evident among sao praphet song-who are both sexually oriented towards men and markedly feminine. In birth-assigned females, there were no clear sexual orientation effects, but there were possible gender-related effects. Groups of individuals who tend to be more masculine (i.e., toms, lesbians) showed more male-typical patterns on weight and leg length than some groups of individuals who tend to be less masculine (i.e., heterosexual women, dees). Thus, it appears the various anthropometrics inform separate biodevelopmental processes that differentially relate to sexual orientation and gender identity/expression depending on the measure in question as well as birth-assigned sex.

Facial Attractiveness of Sisters of Thai Gay Men and Sao Praphet Song.

Androphilia refers to sexual attraction toward adult males. Androphilic males' female genetic relatives might offset the fitness cost of androphilia by having elevated numbers of offspring. Increased attractiveness relative to other women may enable these females to marry up the social hierarchy, providing greater access to resources to support more offspring. In Thailand, there is a marked status hierarchy and male androphiles are highly visible - identifying as gay men or as a third gender known as sao praphet song. Thus, a field experiment was conducted in which 208 heterosexual men rated the facial attractiveness of 25 sisters of Thai gay men or sao praphet song and 25 comparison women with no known same-sex attracted relatives. Contrary to the prediction, the comparison group was rated as more attractive than the sisters, casting doubt on hypotheses that posit greater attractiveness of female relatives of male androphiles as a proximate mechanism facilitating the evolutionary maintenance of genes influencing male androphilia. Also, men from rural areas rated the women, regardless of group, as more attractive than men from urban areas, consistent with some prior studies showing rural-urban differences in perceptions of attractiveness.

Direct Reproduction and Sexual Orientation and Gender Diversity in Thailand.

Human same-sex sexual attraction is considered to be an evolutionary paradox. This paradox rests on same-sex attracted individuals having lowered direct reproduction, indicating reduced direct fitness of genes that influence same-sex attraction. Yet, relatively few empirical studies have examined the relation between same-sex sexual attraction and direct reproduction. The present study examined direct reproduction (i.e., number of children produced) in a diverse Thai sample (N = 1623) of heterosexual men and women, gay and bisexual men, and lesbian and bisexual women, as well as unique Thai gender categories: sao praphet song (i.e., feminine androphilic third-gender males), toms (i.e., masculine gynephilic third-gender females), and dees (i.e., feminine females who are sexually attracted to toms). In males, heterosexual men were more likely to have produced children than other groups, especially as they aged, and none of the gay men or sao praphet song reported having children. In females, there was some degree of reproduction in each of the groups considered. On average, heterosexual and bisexual women had similar numbers of children, whereas heterosexual women had significantly more children than lesbian women, toms, and dees. Number of children was significantly positively associated with age among heterosexual women, bisexual women, toms, and dees. With age, the discrepancies in average number of children produced between heterosexual women and each of lesbian women and toms became significantly more pronounced. Discussion focuses on how these findings, in conjunction with those of prior studies, inform our understanding of evolutionary selection pressures related to human same-sex sexuality.

Handedness and Birth Order Among Heterosexual Men, Gay Men, and Sao Praphet Song in Northern Thailand.

Previous research has examined handedness and birth order to inform sexual orientation and gender identity/role expression development; however, sexual orientation and gender identity/role expression have rarely been disentangled to provide a more nuanced perspective. In Thailand, we investigated sexual orientation and gender identity simultaneously via comparison of 282 heterosexual men, 201 gay men, and 178 sao praphet song-i.e., androphilic, markedly feminine males recognized as a "third" gender. Handedness was examined as: extremely left-handed, moderately left-handed, ambidextrous, moderately right-handed, or extremely right-handed. Birth order was examined as numbers of older and younger brothers and sisters, by using Berglin's, fraternal, and sororal indices, and by examining the older brother odds ratio and sibling sex ratio. Compared with heterosexual men, gay men and sao praphet song were more likely to be extremely right-handed. Sao praphet song were also more likely to be extremely left-handed than heterosexual and gay men. Heterosexual men and sao praphet song had later sororal birth order compared with the expected Thai population value, suggesting stopping rules influenced when probands' mothers ceased having children. These findings provide new insights and replicate previous findings in a non-Western sample. Regarding handedness, in males, mechanisms related to extreme right-handedness likely influence the development of androphilia, whereas mechanisms related to both extreme right- and extreme left-handedness likely explain the combination of androphilia and feminine gender identity/role expression. Regarding birth order, similar to the conclusions of some prior research, stopping rules pose a challenge for testing the fraternal birth order effect.

Evidence for distinct biodevelopmental influences on male sexual orientation.

Several biological mechanisms have been proposed to influence male sexual orientation, but the extent to which these mechanisms cooccur is unclear. Putative markers of biological processes are often used to evaluate the biological basis of male sexual orientation, including fraternal birth order, handedness, and familiality of same-sex sexual orientation; these biomarkers are proxies for immunological, endocrine, and genetic mechanisms. Here, we used latent profile analysis (LPA) to assess whether these biomarkers cluster within the same individuals or are present in different subgroups of nonheterosexual men. LPA defined four profiles of men based on these biomarkers: 1) A subgroup who did not have these biomarkers, 2) fraternal birth order, 3) handedness, and 4) familiality. While the majority of both heterosexual and nonheterosexual men were grouped in the profile that did not have any biomarker, the three profiles associated with a biomarker were composed primarily of nonheterosexual men. We then evaluated whether these subgroups differed on measures of gender nonconformity and personality that reliably show male sexual orientation differences. The subgroup without biomarkers was the most gender-conforming whereas the fraternal birth order subgroup was the most female-typical and agreeable, compared with the other profiles. Together, these findings suggest there are multiple distinct biodevelopmental pathways influencing same-sex sexual orientation in men.

Both neural and global androgen receptor overexpression affect sexual dimorphism in the mouse brain.

Testosterone is the main endocrine mechanism mediating sexual differentiation of the mammalian brain, although testosterone signalling is complex and important mechanistic questions remain. Notably, the extent to which testosterone acts via androgen receptors (AR) in this process remains unknown and it is also not clear where testosterone acts in the body to produce sexual dimorphisms in neuroanatomy. To address these questions, we used a transgenic mouse model of Cre/loxP-driven AR overexpression in which AR was induced selectively in neural tissue (Nestin-cre) or in all tissues (CMV-cre). We then studied sexually dimorphic features of several well-characterised sexual dimorphisms: calbindin-immunoreactive neurones in the medial preoptic area (CALB-SDN), tyrosine hydroxylase neurones in the anteroventral periventricular nucleus, and vasopressin-immunoreactive neurones originating in the bed nucleus of the stria terminalis and their projections in the lateral septum. We additionally evaluated oestrogen receptor α immunoreactivity in these nuclei. Briefly, we found that global but not neural overexpression of AR resulted in masculinisation of CALB-SDN nucleus volume, cell number and cell size in transgenic females. Furthermore, neural AR overexpression resulted in increased oestrogen receptor α staining in females compared to males in the medial preoptic area. AR overexpression did not affect other measures. Overall, the results of the present study provide support for the hypothesis that androgenic mechanisms external to the nervous system can affect sexual differentiation of the brain.

Gender Nonconformity and Birth Order in Relation to Anal Sex Role Among Gay Men.

Androphilia is associated with an elevated number of older brothers among natal males. This association, termed the fraternal birth order effect, has been observed among gay men who exhibit marked gender nonconformity. Gender nonconformity has been linked to gay men's preferred anal sex role. The present study investigated whether these two lines of research intersect by addressing whether the fraternal birth order effect was associated with both gender nonconformity and a receptive anal sex role (243 gay men, 91 heterosexual men). Consistent with previous research, we identified the fraternal birth order effect in our sample of gay men. Also, gay men were significantly more gender-nonconforming on adulthood and recalled childhood measures compared to heterosexual men. When gay men were compared based on anal sex role (i.e., top, versatile, bottom), all groups showed significantly greater recalled childhood and adult male gender nonconformity than heterosexual men, but bottoms were most nonconforming. Only gay men with a bottom anal sex role showed evidence of a fraternal birth order effect. A sororal birth order effect was found in our sample of gay men, driven by versatiles. No significant associations were found between fraternal birth order and gender nonconformity measures. These results suggest that the fraternal birth order effect may apply to a subset of gay men who have a bottom anal sex role preference and that this subgroup is more gender-nonconforming. However, there were no significant associations between fraternal birth order and gender nonconformity at the individual level. As such, based on the present study, whether processes underpinning the fraternal birth order effect influence gender nonconformity is equivocal.

Handedness is a biomarker of variation in anal sex role behavior and Recalled Childhood Gender Nonconformity among gay men.

Developmental theories of the biological basis of sexual orientation suggest that sexually differentiated psychological and behavioural traits should be linked with sexual orientation. Subgroups of gay men delineated by anal sex roles differ according to at least one such trait: gender expression. The present study assessed the hypothesis that handedness, a biologically determined sexually differentiated trait, corresponds to differences in subgroups of gay men based on anal sex role. Furthermore, it assessed whether handedness mediates the association between gender nonconformity and male sexual orientation. Straight and gay men (N = 333) completed the Edinburgh Inventory of Handedness and the Recalled Childhood Gender Nonconformity Scale. Gay men also completed measures of anal sex role preference. As in previous studies, gay men showed greater non-right-handedness and gender nonconformity than straight men. Also, among gay men, bottoms/versatiles (i.e., gay men who take a receptive anal sex role, or who take on both a receptive and insertive anal sex role) were more gender-nonconforming than tops (i.e., gay men who take an insertive anal sex role). In support of the hypothesis, bottoms/versatiles were more non-right-handed than tops and handedness mediated the male sexual orientation and anal sex role differences in Recalled Childhood Gender Nonconformity. Together, these findings suggest that developmental processes linked to handedness underpin variation among men in sexual orientation and gender nonconformity as well as variation among subgroups of gay men that are delineated by anal sex roles.

Nonneural Androgen Receptors Affect Sexual Differentiation of Brain and Behavior.

Testosterone, acting via estrogenic and androgenic pathways, is the major endocrine mechanism promoting sexual differentiation of the mammalian nervous system and behavior, but we have an incomplete knowledge of which cells and tissues mediate these effects. To distinguish between neural and nonneural actions of androgens in sexual differentiation of brain and behavior, we generated a loxP-based transgenic mouse, which overexpresses androgen receptors (ARs) when activated by Cre. We used this transgene to overexpress AR globally in all tissues using a cytomegalovirus (CMV)-Cre driver (CMV-AR), and we used a Nestin-Cre driver to overexpress AR only in neural tissue (Nes-AR). We then examined whether neural or global AR overexpression can affect socio-sexual behaviors using a resident-intruder paradigm. We found that both neural and global AR overexpression resulted in decreased aggressive behaviors and increased thrusting during mounting of intruders, consistent with a neural site of action. Global, but not neural, AR overexpression in males led to an increase in same-sex anogenital investigation. Together, these results suggest novel roles for nonneural AR in sexual differentiation of mice, and indicate that excess AR can lead to a paradoxical reduction of male-typical behavior.