In vitro metabolism of progesterone, androgens and estrogens by rainbow trout embryos

The purpose of the study was to determine the steroid metabolic pathways used by rainbow trout (Oncorhynchus mykiss) embryos. Whole embryo preparations at 42, 53 and 65 days post fertilization (dpf) were incubated, in vitro, with the tritium-labelled steroids, progesterone (P₄), testosterone (T), androstenedione (A4), 17β-estradiol (E₂), and estrone (E₁), and the metabolites formed were separated and identified by reverse phase high performance liquid chromatography (HPLC). The degree of metabolism for each of the substrates was dependent on the age of the embryos, and was always higher in older embryos. P₄ yielded three metabolites, one of them identified by gas chromatography-mass spectroscopy as 3β,7α-dihydroxy-5α-pregnan-20-one. Several unknown metabolites of a similar profile on HPLC were produced for T and A₄, in addition to A₄ and T as metabolites of T and A ₄ metabolism, respectively. Likewise, E₂ and E₁ yielded each other and an unknown polar metabolite. These observations argue for the presence of 5α-reductase, 7α-hydroxylase and 17β-hydroxysteroid dehydrogenase activity in embryo tissues which, by their conversion of biologically active yolk steroids of maternal origin, may provide a protective environment for the embryo during early development.