Stage-related changes in steroid-converting enzyme activities in Squalus testis: synthesis of biologically active metabolites via 3 beta-hydroxysteroid dehydrogenase/isomerase and 5 alpha-reductase.

To investigate the relationship between steroidogenesis and spermatogenesis, two key enzymes of androgen biosynthesis, 3 beta-hydroxysteroid dehydrogenase/isomerase (3 beta-HSD) and 5 alpha-reductase, were compared at premeiotic (PrM), meiotic (M), and postmeiotic (PoM) stages. Staged tissues were obtained by dissection from the testis of the spiny dogfish Squalus acanthias, in which spermatogenesis is a simple diametric progression. Microsomal 3 beta-HSD activity was measured by conversion of [3H]dehydroepiandrosterone (DHEA) to androstenedione (AE). Reaction constants were: Km = 3.2 microM and Vmax = 243 pmol/min/mg protein. 3 beta-HSD increased progressively with maturation, resulting in three- to four-fold higher levels in PoM than in PrM stages. Absolute values and stage-related differences were the same, whether microsomes were derived from whole testis or from isolated spermatocysts (germ cell/Sertoli cell units), thus supporting microscopic studies showing that Sertoli cells are the primary steroidogenic elements of dogfish testis. In vitro conversion of [3H]testosterone to [3H]dihydrotestosterone (DHT) was used to estimate 5 alpha-reductase activity. Apparent substrate affinity was similar to that of 3 beta-HSD (Km = 2.9 microM), but maximal product yields were two to three orders of magnitude lower (Vmax = 208 fmol/min/mg protein). Also, the stage-related pattern of 5 alpha-reductase activity (PrM > PoM >> M) differed from that of 3 beta-HSD (PoM >> M > PrM).(ABSTRACT TRUNCATED AT 250 WORDS)