Sex steroids were measured by radioimmunoassay in whole-body extracts of coho salmon, Oncorhynchus kisutch, during early development and sexual differentiation. Profiles were developed for fish from the time of fertilization until 87 days postfertilization (dpf) for six steroids: testosterone (T), 11-ketotestosterone (KT), androstenedione (A), progesterone (P4), 17 alpha-hydroxy-20 beta-dihydroprogesterone (DHP), and 17 beta-estradiol (E2). Ovarian fluid was also examined for steroid content. Steroid profiles of unfertilized eggs essentially paralleled those of ovarian fluid. In one experiment, steroids in developing embryos declined precipitously after fertilization until 30 dpf; at hatching, all steroids increased slightly and then declined during yolk sac absorption. Results from a second experiment basically supported those of the first except that only testosterone increased at the time of hatching. Bimodality was evident in the data on steroid levels for fish collected between 42 and 56 dpf and again after 87 dpf. The hormone levels generally decreased or remained constant after the onset of exogenous feeding. Histological analyses during the first experiment showed the presence of undifferentiated gonads between hatching and 70 dpf, but by 77 dpf ovarian development was evident. In the second experiment, in which fish were more frequently sampled for histological analysis, undifferentiated gonads were present from hatching to 59 dpf. Development of oogonia was observed between 66 and 73 dpf and by 75 dpf ovarian development could be easily discerned. The sex of fish sampled at 101 dpf was determined by examining gonadal morphology, and steroid levels of those fish were determined. A sexual dimorphism was apparent in levels of T, KT, and A, but not of DHP or E2. The dynamics of steroid content of developing coho salmon at hatch, coupled with their bimodal distributions during yolk sac absorption, may suggest a role of sex steroids in the process of sexual differentiation apparent later in development. Changes in whole-body steroid levels at hatch may also be indicative of the onset of sexual differentiation even though no signs of gondal differentiation were histologically discernible at that time.