Gonads of Poecilia latipinna transformed [3H]testosterone into a number of reduced and conjugated metabolites in high yield in vitro. In the male 5 beta-androstane-3 alpha, 17 beta-diol, 5 beta-androstane-3 beta, 17 beta-diol, and their "sulphates" were identified. The only 11-oxygenated androgen detected was a compound tentatively identified as 5 beta-androstane-3 beta, 11 beta, 17 beta-triol. In ovarian incubates androstenedione, 5 beta-androstane-3 alpha, 17 beta-diol and its glucuronide, testosterone glucuronide, and 5 beta-androstane-3 alpha, 17 beta-diol glucuronide were identified. Highest yields of the ovarian glucuronides coincided with the termination of vitellogenesis which may indicate a possible pheromonal role of these conjugates. In vivo plasma levels of estradiol in the female were correlated with vitellogenesis and fell markedly after castration or hypophysectomy. In males the plasma concentrations of testosterone, 11-ketotestosterone and 11 beta-hydroxytestosterone and their conjugates were variable and not apparently correlated with testicular weight, but they were reduced to undetectable levels by castration and hypophysectomy. The results suggest that 5 alpha- and 5 beta-reduced steroids may play a role in the reproductive endocrinology of P. latipinna and that measurements of only the "classical" steroid hormones in this and possibly other species may give only a partial and misleading picture of endocrine changes.