Skip to main content
Log in

Synthesis and regulation of 17α-hydroxy-20β-dihydroprogesterone in immature males of Oncorhynchus mykiss

  • Published:
Fish Physiology and Biochemistry Aims and scope Submit manuscript

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References cited

  • Arai, R. and Tamaoki, B. 1967. Steroid biosynthesis in vitro by testes of rainbow trout, Salmo gairdneri. Gen. Comp. Endocrinol. 8: 305–313.

    Google Scholar 

  • Axelrod, L.R., Matthijssen, C., Goldzieher, J.W. and Pulliam, J.E. 1965. Definitive identification of microquantities of radioactive steroids by recrystallization to constant specific activity. Acta Endocrinol. Suppl. 99: 3–67.

    Google Scholar 

  • Baynes, S.M. and Scott, A.P. 1985. Seasonal variations in parameters of milt production and in plasma concentration of sex steroids of male rainbow trout (Salmo gairdneri). Gen. Comp. Endocrinol. 57: 150–160.

    Google Scholar 

  • Billard, R. and Escaffre, A.M. 1975. Identification des stades de la spermatogénèse de la truite fario d'après la morphologie des gonades et la spermiation. Bull. Franç. Piscicult. 256: 111–115.

    Google Scholar 

  • Breton, B., Kann, G., Burzawa-Gerard, E. and Billard, R. 1971. Dosage radioimmunologique d'une hormone gonadotrope de carpe (Cyprinus carpio L.). C.R. Acad. Sci., Paris 272: 1515–1517.

    Google Scholar 

  • Breton, B., Jalabert, B. and Reinaud, P. 1976. Purification of gonadotropin from rainbow trout (Salmo gairdneri Richardson) pituitary glands. Ann. Biol. Anim. Bioch. Biophys. 16: 25–36.

    Google Scholar 

  • Campbell, S.M.C. and Wiebe, J.P. 1989. Stimulation of spermatocyte development in prepubertal rats by the Sertoli cell steroid, 3α-hydroxy-4-pregnen-20-one. Biol. Reprod. 40: 897–905.

    Google Scholar 

  • Crim, L.W., Peter, R.E. and Billard, R. 1981. Onset of gonadotropic hormone accumulation in the immature trout pituitary gland in response to estrogen or aromatizable androgen steroid hormones. Gen. Comp. Endocrinol. 44: 374–381.

    Google Scholar 

  • Crim, L.W., Billard, R., Genge, P.D. and Idler, D.R. 1982. The influence of immature gonads on onset of gonadotropic hormone accumulation in the juvenile rainbow trout pituitary gland. Gen. Comp. Endocrinol. 48: 161–166.

    Google Scholar 

  • Déphe, J. and Sire, O. 1982. In vitro metabolism of progesterone and 17α-hydroxyprogesterone in the testis of the rainbow trout, Salmo gairdneri Rich., at different stages of spermatogenesis. Reprod. Nutr. Dev. 22: 427–438.

    Google Scholar 

  • Fostier, A. and Jalabert, B. 1986. Steroidogenesis in rainbow trout (Salmo gairdneri) at various preovulatory stages: changes in plasma hormone levels and in vivo and in vitro responses of the ovary to salmon gonadotropin. Fish Physiol. Biochem. 2: 87–99.

    Google Scholar 

  • Gane, M. 1968. Techniques Histologiques. Masson Eds, Paris.

    Google Scholar 

  • Groves, D.J. and Batten, T.F.C. 1986. Direct control on the gonadotroph in a teleost. Poecilla latipinna: gonadal steroids. Gen. Comp. Endocrinol. 61: 402–416.

    Google Scholar 

  • Kobayashi, M., Aida, K. and Hanyu, I. 1986. Effects of HCG on milt amount and plasma levels of steroid hormones in male goldfish. Bull. Jap. Soc. Sci. Fish. 52: 755.

    Google Scholar 

  • Le Bail, P.Y., Fostier, A. and Marcuzzi, O. 1983. Limites et améliorations du sexage des salmonidés par dosage de la 11-cétotestostérone circulante. Can. J. Zool. 61: 457–460.

    Google Scholar 

  • Le Gac, F. and Loir, M. 1988. Control of testis function in fish: in vitro studies of gonadotropic regulation in the trout (Salmo gairdneri). Reprod. Nutr. Dévelop. 28: 1031–1046.

    Google Scholar 

  • Loir, M. 1988. Trout Sertoli and Leydig cells: isolation, separation, and culture. Gamete Res. 20: 437–458.

    Google Scholar 

  • Loir, M. 1989. Trout Sertoli cells and germ cells in primary culture: I. Morphological and ultrastructural study. Gamete Res. 24: 151–169.

    Google Scholar 

  • Loir, M. 1990a. Trout steroidogenic testicular cells in primary culture. I. Changes in free and conjugated androgen and progestagen secretions: effects of gonadotropin, serum, and lipoproteins. Gen. Comp. Endocrinol. 78: 374–387.

    Google Scholar 

  • Loir, M. 1990b. Trout steroidogenic testicular cells in primary culture. II. Steroidogenic activity of interstitial cells, Sertoli cells, and spermatozoa. Gen. Comp. Endocrinol. 78: 388–398.

    Google Scholar 

  • Loir, M. 1990c. Interstitial cells from the testis of the trout (Oncorhynchus mykiss) in vivo and in primary culture. Cell Tiss. Res. 261: 133–144.

    Google Scholar 

  • Lou, S.W., Aida, K., Hanyu, I., Sakai, K., Nomura, M., Tanaka, M. and Takazi, S. 1986. Endocrine profiles in the males of a twice-annually spawning strain of rainbow trout, Salmo gairdneri Gen. Comp. Endocrinol. 64: 212–219.

    Google Scholar 

  • Miwa, S., Yan, L. and Swanson, P. 1994. Localization of two gonadotropin receptors in the salmon gonad by in vitro ligand autoradiography. Biol. Reprod. 50: 629–642.

    Google Scholar 

  • Nagahama, Y., Young, G., Ueda, H., Kagawa, H. and Adachi, S. 1983. Endocrine control of final maturation in salmonids. In Salmonid Reproduction. pp. 9–19. Edited by R.N. Iwamoto and S. Sower. Seattle.

  • Nagahama, Y. 1987. Gonadotropin action on gametogenesis and steroidogenesis in teleost gonads. Zool. Sci. 4: 209–222.

    Google Scholar 

  • Oota, I. and Yamamoto, K. 1966. Interstitial cells in the immature testes of the rainbow trout. Ann. Zool. Jap. 39: 142–148.

    Google Scholar 

  • Pankhurst, N.W., Van der Kraak, G., Peter, R.E. and Breton, B. 1986. Effects of (D-Ala6, Pro9N ethylamide)-LHRH on plasma levels of gonadotropin, 17α,20β-dihydroxy-4-pregnen-3-one and testosterone in males goldeye (Hiodon alasoides Rafineque). Fish Physiol. Biochem. 1: 163–170.

    Google Scholar 

  • Planas, J.V., Swanson, P. and Dickhoff, W.W. 1991. Regulation of testicular steroidogenesis by coho salmon gonadotropins, GtH I and GtH II, in vitro. In Proc. IVth Int. Symp. Reproductive Physiology of Fish. p. 103. University of East Anglia, Norwich.

    Google Scholar 

  • Planas, J.V., Swanson, P. and Dickhoff, W.W. 1993. Regulation of testicular steroid production in vitro by gonadotropins (GtH I and GtH II) and cyclic AMP in coho salmon (Oncorhynchus kisutch). Gen. Comp. Endocrinol. 91: 8–24.

    Google Scholar 

  • Sakai, N., Ueda, H., Suzuki, N. and Nagahama, Y. 1989a. Involvement of sperm in the production of 17α,20β-dihydroxy-4-pregnen-3-one in the testis of spermiating rainbow trout, Salmo gairdneri. Biomed. Res. 10: 131–138.

    Google Scholar 

  • Sakai, N., Ueda, H., Suzuki, N. and Nagahama, Y. 1989b. Steroid production by Amago salmon (Oncorhynchus rhodurus) testes at different developmental stages. Gen. Comp. Endocrinol. 75: 231–240.

    Google Scholar 

  • Sangalang, G.B. and Freeman, H. 1988. In vitro biosynthesis of 17α,20β-dihydroxy-4-pregnen-3-one by the ovaries, testes, and head kidneys of the Atlantic salmon Salmo salar. Gen. Comp. Endocrinol. 69: 406–415.

    Google Scholar 

  • Schulz, R., Goos, H.J.Th. and Blüm, V. 1988. Salmon gonadotropin (sGtH) immunoreactivity and 11-oxotestosterone secretion of mature rainbow trout (Salmo gairdneri) testes in vitro: an alternative to radio-receptor assay for sGtH-binding studies. Cell Tiss. Res. 251: 665–669.

    Google Scholar 

  • Schulz, R. and Blüm, V. 1990. Steroid secretion of rainbow trout testis in vitro: variation during the reproductive cycle. Gen. Comp. Endocrinol. 80: 1–10.

    Google Scholar 

  • Schulz, R.W., Andriske, M., Lembke, P.J. and Blüm, V. 1992. Effect of salmon gonadotropin hormone on sex steroids in male rainbow trout: plasma levels and testicular secretion in vitro. J. Comp. Physiol. B. 162: 224–230.

    Google Scholar 

  • Scott, A.P. and Sumpter, J.P. 1989. Seasonal variation in testicular germ cell stages and in plasma concentrations of sex steroids in male rainbow trout (Salmo gairdneri) maturing at 2 years old. Gen. Comp. Endocrinol. 73: 46–58.

    Google Scholar 

  • Schwartz, D. 1963. Méthodes Statistiques à l'Usage des Médecins et des Biologistes. Flamarion Médecine-Sciences, Paris.

    Google Scholar 

  • Simpson, T.H., Wright, R.S. and Hunt, S.V. 1964. Steroid biosynthesis in the testis of dogfish (Squalus acanthias). J. Endocrinol. 31: 29–38.

    Google Scholar 

  • Swanson, P., Bernard, M., Nozaki, M., Suzuki, K., Kawauchi, H. and Dickhoff, W.W. 1989. Gonadotropins I and II in juvenile coho salmon. Fish Physiol. Biochem. 7: 169–176.

    Google Scholar 

  • Swanson, P. 1991. Salmon gonadotropins: reconciling old and new ideas. In Proc. IVth Int. Symp. Reproductive Physiology of Fish. p. 2–7. University of East Anglia, Norwich.

    Google Scholar 

  • Ueda, H., Young, G., Crim, L.W., Kambegawa, A. and Nahagama, Y. 1983. 17α,20β-Dihydroxy-4-pregnen-3-one: plasma levels during sexual maturation and in vitro production by testes of amago salmon (Oncorhynchus rhodurus) and rainbow trout (Salmo gairdneri). Gen. Comp. Endocrinol. 51: 106–112.

    Google Scholar 

  • Ueda, H., Kambegawa, A. and Nagahama, Y. 1984. In vitro 11-ketotestosterone and 17α,20β-dihydroxy-4-pregnen-3-one production by testicular fragments and isolated sperm of rainbow trout, Salmo gairdneri. J. Exp. Zool. 231: 435–439.

    Google Scholar 

  • Ueda, H., Kambegawa, A. and Nagahama, Y. 1985. Involvement of gonadotrophin and steroid hormones in spermiation in the amago salmon, Oncorhynchus rhodurus, and goldfish, Carassius auratus. Gen. Comp. Endocrinol. 59: 24–30.

    Google Scholar 

  • Van den Hurk, R., Lambert, J.G.D. and Peute, J. 1982. Steroidogenesis in the gonads of rainbow trout fry (Salmo gairdneri) before and after the onset of gonadal sex differentiation. Reprod. Nutr. Dévelop. 22: 413–425.

    Google Scholar 

  • Vizziano, D. 1993. Production de la 17α-hydroxy-20β-dihydro-progestérone par le testicule de truite arc en ciel (Oncorhynchus mykiss) à différents stades de la spermatogénèse. Aspects de sa régulation et de sa localisation cellulaire. PhD Thesis, Université de Rennes I. France.

    Google Scholar 

  • Vizziano, D., Fostier, A., Le Gac, F. and Loir, M. 1995. Localization of 20β-hydroxysteroid dehydrogenase activity in non-flagellated germ cells of rainbow trout testis. Biol. Reprod. (In press).

  • Weil, C. and Marcuzzi, O. 1990. Cultured pituitary cell GtH response to GnRh at different stages of rainbow trout spermatogenesis and influence of steroid hormones. Gen. Comp. Endocrinol. 79: 492–498.

    Google Scholar 

  • Yan, L., Swanson, P. and Dickhoff, W.W. 1991. Binding of gonadotropins (GtH I and GtH II) to coho salmon gonadal membrane preparations. J. Exp. Zool. 258: 221–230.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Denise Vizziano.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vizziano, D., Le Gac, F. & Fostier, A. Synthesis and regulation of 17α-hydroxy-20β-dihydroprogesterone in immature males of Oncorhynchus mykiss . Fish Physiol Biochem 14, 289–299 (1995). https://doi.org/10.1007/BF00004067

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00004067

Keywords

Navigation