The influence of hormones on the developing fetus has been the most influential causal hypothesis of the development of sexual orientation. In simple terms, the developing fetal brain begins in a "female" typical state. The presence of the Y-chromosome in males prompts the development of testes, which release testosterone, the primary androgen receptor-activating hormone, to masculinize the fetal brain. This masculinizing effect pushes males towards male typical brain structures, and most of the time, attraction to females. It has been hypothesized that
gay men may have been exposed to lower levels of testosterone in key regions of the brain, or had different levels of receptivity to its masculinizing effects, or experienced fluctuations at critical times. In women, it is hypothesized that high levels of exposure to testosterone in key regions may increase likelihood of same sex attraction.).
Lesbians on average were found to have significantly more masculine digit ratios, a finding which has been replicated numerous times in studies cross-culturally, but sometimes contradicted, such as when ethnicity is taken into account. While direct effects are hard to measure for ethical reasons, animal experiments where scientists manipulate exposure to sex hormones during gestation can also induce life long male-typical behavior and mounting in female animals, and female-typical behavior in male animals. Maternal immune responses during fetal development are strongly demonstrated as causing male homosexuality and bisexuality. Research since the 1990s has demonstrated that the more male sons a woman has, there is a higher chance of later born sons being gay. During pregnancy, male cells enter a mother's bloodstream, which are foreign to her immune system. In response, she develops antibodies to neutralize them. These antibodies are then released on future male fetuses and may neutralize Y-linked antigens, which play a role in brain masculinization, leaving areas of the brain responsible for sexual attraction in the female-typical position, or attracted to men. The more sons a mother has will increase the levels of these antibodies, thus creating the observed
fraternal birth order effect. Biochemical evidence to support this effect was confirmed in a lab study in 2017, finding that mothers with a gay son, particularly those with older brothers, had heightened levels of antibodies to the
NLGN4Y Y-protein than mothers with heterosexual sons.
J. Michael Bailey has described maternal immune responses as "causal" of male homosexuality. This effect is estimated to account for between 15-29% of gay men, while other gay and bisexual men are thought to owe sexual orientation to genetic and hormonal interactions. The size of the sheep oSDN has also been demonstrated to be formed in utero, rather than postnatally, underscoring the role of prenatal hormones in masculinization of the brain for sexual attraction. Rice et al. propose that these epimarks normally canalize sexual development, preventing
intersex conditions in most of the population, but sometimes failing to erase across generations and causing reversed sexual preference. Testing this hypothesis is possible with current stem cell technology. There is evidence that mutations in NLGN4X and NLGN4Y are linked to autism spectrum conditions and such conditions may be elevated in asexual people. Thus, NLGN4X/Y may affect neurological functioning associated with, broadly, the forming of social connections to others, including sexual/romantic ones. == Childhood gender nonconformity ==