Effect of gonadotropin type II and 17-hydroxy-4-pregnene-3,20-dione on 17,20β-dihydroxy-4-pregnen-3-one production by rainbow trout testes at different developmental stages
Three experimental approaches were chosen to study the question if the progestin 17α-hydroxy-20β-dihydroprogesterone (17α20βOHP) is synthesised in testes of young Oncorhynchus mykiss, in which the absence of spermatozoa was verified histologically: first, in order to detect 20β-hydroxysteroid dehydrogenase activity (20βHSD), testes homogenates were incubated with 3H-labeled 17αOHP.Metabolites were analysed by TLC, HPLC, and repeated crystallization to constant isotope ratios. One of the metabolites was identified as 17α20βOHP-3H, indicating that already immature testes contain 20βHSD activity and are able to produce 20β-reduced steroids. Second, 17α20βOHP was quantified by radioimmunoassay in incubates of testes fragments. The sensitivity of the gonads to gonadotropin II (GtH II) became evident when comparing incubations in the absence and presence of GtH II. Third, plasma levels of 17α20βOHP were significantly higher in animals injected with partially purified salmon gonadotropin, compared to controls. Thus, for the first time, it could be shown that 20βHSD is present in testicular cells other than spermatozoa. Furthermore, 17α20βOHP is indeed secreted at a very early stage of testicular development; 17α20βOHP secretion is also responsive to GtH II. Future studies will have to show if the functions of this progestin include the stimulation of spermatogenesis.