The current study tested the hypothesis that elevated childhood separation anxiety is associated with female-typical childhood behavior and identity by comparing retrospective reports of heterosexual and homosexual men and women (N = 399). Participants completed measures of recalled childhood separation anxiety and childhood gender-atypical behavior and identity. Heterosexual men reported significantly less childhood separation anxiety relative to all other groups. Childhood gender atypicality was significantly positively correlated with childhood separation anxiety among homosexual men, but not among members of other participant groups. Discussion focused on the implications of these findings for the proposed hypothesis as well as future directions for research examining the bases of developmental associations among sex, sexual orientation, gender atypicality, and childhood separation anxiety.