To better understand sexual orientation from an evolutionary perspective, we investigated whether, compared to heterosexual men, the fewer direct descendants of homosexual men could be counterbalanced by a larger number of other close biological relatives. We also investigated the extent to which three patterns generally studied separately--handedness, number of biological older brothers, and hair-whorl rotation pattern--correlated with each other, and for evidence of replication of previous findings on how each pattern related to sexual orientation. We surveyed at Gay Pride and general community festivals, analyzing data for 894 heterosexual men and 694 homosexual men, both groups predominantly (*80%) white/non-Hispanic. The Kinsey distribution of sexual orientation for men recruited from the general community festivals approximated previous population-based surveys. Compared to heterosexual men, homosexual men had both more relatives, especially paternal relatives, and more homosexual male relatives. We found that the familiality for male sexual orientation decreased with relatedness, i.e., when moving from first-degree to seconddegree relatives. We also replicated the fraternal birth order effect. However, we found no significant correlations among handedness, hair whorl rotation pattern, and sexual orientation, and, contrary to some previous research, no evidence that male sexual orientation is transmitted predominantly through the maternal line.

Results, especially in the context of past studies, support the existence of genes on pericentromeric chromosome 8 and chromosome Xq28 influencing development of male sexual orientation.

Family and twin studies suggest that genes play a role in male sexual orientation. We conducted a genome-wide association study (GWAS) of male sexual orientation on a primarily European ancestry sample of 1,077 homosexual men and 1,231 heterosexual men using Affymetrix single nucleotide polymorphism (SNP) arrays. We identified several SNPs with p < 10−5, including regions of multiple supporting SNPs on chromosomes 13 (minimum p = 7.5 × 10−7) and 14 (p = 4.7 × 10−7). The genes nearest to these peaks have functions plausibly relevant to the development of sexual orientation. On chromosome 13, SLITRK6 is a neurodevelopmental gene mostly expressed in the diencephalon, which contains a region previously reported as differing in size in men by sexual orientation. On chromosome 14, TSHR genetic variants in intron 1 could conceivably help explain past findings relating familial atypical thyroid function and male homosexuality. Furthermore, skewed X chromosome inactivation has been found in the thyroid condition, Graves’ disease, as well as in mothers of homosexual men. On pericentromeric chromosome 8 within our previously reported linkage peak, we found support (p = 4.1 × 10−3) for a SNP association previously reported (rs77013977, p = 7.1 × 10−8), with the combined analysis yielding p = 6.7 × 10−9, i.e., a genome-wide significant association.

Male sexual orientation is influenced by environmental and complex genetic factors. Childhood gender nonconformity (CGN) is one of the strongest correlates of homosexuality with substantial familiality. We studied brothers in families with two or more homosexual brothers (409 concordant sibling pairs in 384 families, as well as their heterosexual brothers), who self-recalled their CGN. To map loci for CGN, we conducted a genome-wide linkage scan (GWLS) using SNP genotypes. The strongest linkage peaks, each with significant or suggestive two-point LOD scores and multipoint LOD score support, were on chromosomes 5q31 (maximum two-point LOD = 4.45), 6q12 (maximum two-point LOD = 3.64), 7q33 (maximum two-point LOD = 3.09), and 8q24 (maximum two-point LOD = 3.67), with the latter not overlapping with previously reported strongest linkage region for male sexual orientation on pericentromeric chromosome 8. Family-based association analyses were used to identify associated variants in the linkage regions, with a cluster of SNPs (minimum association p = 1.3 × 10–8) found at the 5q31 linkage peak. Genome-wide, clusters of multiple SNPs in the 10–6 to 10–8p-value range were found at chromosomes 5p13, 5q31, 7q32, 8p22, and 10q23, highlighting glutamate-related genes. This is the first reported GWLS and genome-wide association study on CGN. Further increasing genetic knowledge about CGN and its relationships to male sexual orientation should help advance our understanding of the biology of these associated traits.

Gender nonconformity-both in childhood and in adulthood-is a strong correlate of male sexual orientation, with homosexual men showing greater gender nonconformity during both periods. Gender nonconformity also tends to be markedly variable, with some homosexual men being more feminine, or less masculine, than others. We replicated large differences in self-reported childhood and adult gender nonconformity between homosexual and heterosexual men. Subsequently, we examined the extent to which homosexual men's variation in gender nonconformity runs in families by examining pairs of genetic brothers who were either both homosexual (concordant pairs) or had different sexual orientations (discordant pairs). Both concordant and discordant pairs yielded modest positive correlations consistent with moderate genetic and/or familial environmental effects on gender nonconformity. Our results support the feasibility of supplementing genetic studies of male sexual orientation with analyses of gender nonconformity variation.