A variety of reptiles possess temperature-dependent sex determination (TSD) in which the incubation temperature of a developing egg determines the gonadal sex. Current evidence suggests that temperature signals may be transduced into steroid hormone signals with estrogens directing ovarian differentiation. Steroidogenic factor 1 (SF-1) is one component of interest because it regulates the expression of steroidogenic enzymes in mammals and is differentially expressed during development of testis and ovary. Northern blot analysis of SF-1 in developing tissues of the red-eared slider turtle (Trachemys scripta), a TSD species, detected a single primary SF-1 transcript of approximately 5.8 kb across all stages of development examined. Analysis by in situ hybridization indicated nearly equivalent SF-1 expression in early, bipotential gonads at male (26°C)- and female (31°C)-producing incubation temperatures. In subsequent stages, as gonadal sex first becomes histologically distinguishable during the temperature-sensitive period, SF-1 expression increased in gonads at a male-producing temperature and decreased at a female-producing temperature, suggesting a role for SF-1 in the sex differentiation pathway. SF-1 message was also found in adrenal and in the periventricular region of the preoptic area and diencephalon, but there was no apparent sex bias in these tissues at any stage examined. The overall developmental pattern of SF-1 mRNA expression in T. scripta appears to parallel that found in mammals, indicating possible homologous functions.