A neuroendocrine component, the positive estrogen feedback effect, thought to be related to sexual orientation and, indirectly, to sexual differentiation, was evaluated in healthy, noninstitutionalized research volunteers. Men and women with a lifelong heterosexual orientation and men with a lifelong homosexual orientation were administered an estrogen preparation known to enhance the concentration of luteinizing hormone in women but not in men. The secretory pattern of luteinizing hormone in the homosexuals in response to estrogen was intermediate between that of the heterosexual men and that of the women. Furthermore, testosterone was depressed for a significantly longer period in the homosexual men than in the heterosexual men. These findings suggest that biological markers for sexual orientation may exist.