Summary
The fertilizing ability of spermatozoa from epididymal tubules maintained in organ cultures from 1 to 7 days was assessed after artificial insemination into receptive does. It was found that spermatozoa from the distal corpus which were already capable of fertilizing eggs prior to the cultures retain this ability for 1 day without addition of hormone and for 3–4 days when testosterone (0.5 μg/ml) or 5α-dihydrotestosterone (0.5 μg/ml) is added to the culture medium. Spermatozoa from the proximal corpus which were not capable of fertilizing eggs prior to the cultures remain so after 1 day in cultures without addition of hormone. Testosterone, 5α-dihydrotestosterone, 3α-androstanediol, or 3β-androstanediol was added to cultures of proximal corpus at a concentration of 0.5 μg/ml. Only with 5α-DHT is the mean percentage of fertilization significantly higher than the percentage obtained without addition of hormone. Insulin does not potentiate the effect of 5α-DHT on sperm fertilizing ability. Epithelial growth factor is ineffective. Spermatozoa from the caput epididymidis kept in cultures for 1 to 4 days remain infertile. The results are discussed in light of the morphological findings presented in the preceding communication and in relation to the physiological requirement for sperm maturation in the epididymis.
Similar content being viewed by others
References
Anderson, K.M., Liao, S.: Selective retention of dihydrotestosterone by prostatic nuclei. Nature(Lond.) 219, 277–279 (1968)
Barthe, P.L., Bullock, L.P., Mowszowicz, I, Bardin, C.W., Orth, D.N.: Submaxillary gland epidermal growth factor: A sensitive index of biologic androgen activity. Endocrinology 95, 1019–1025 (1974)
Baulieu, E.E., Lasnitzki, I., Robel, P.: Metabolism of testosterone and action of metabolites on prostate glands grown in organ culture. Nature (Lond.) 219, 1155–1156 (1968)
Bedford, J.M.: Effects of duct ligation on the fertilizing ability of spermatozoa from different regions of the rabbit epididymis. J. exp. Zool. 166, 271–282 (1967)
Blaquier, J.A.: An in vitro action of androgens on protein synthesis by epididymal tubules maintained in organ culture. Biochem. biophys. Res Commun. 52, 1177–1183 (1973)
Blaquier, J.A.: The influence of androgens on protein synthesis by cultured rat epididymal tubules. Acta endocr. (Kbh.) 79, 403–416 (1975)
Blaquier, J.A., Calandra, R.S: Intranuclear receptor for androgens in rat epididymis. Endocrinology 93, 51–60 (1973)
Bruchovsky, N., Wilson, J.D.: The conversion of testosterone to 5α-androstan-17β-ol-3-one by rat prostate in vivo and in vitro. J. biol. Chem. 243, 2012–2021 (1968a)
Bruchovsky, N., Wilson, J.D.: The intranuclear binding of testosterone and 5α-androstan-17β-ol-3one by rat prostate. J. biol. Chem. 243, 5953–5960 (1968b)
Cohen, D.: Isolation of a mouse submaxillary gland protein accelerating incisor eruption and eyelid opening in the new-born animal. J. biol. Chem. 237, 1555–1562 (1962)
Cohen, S., Savage, C.R., Jr.: Part II. Recent studies on the chemistry and biology of epidermal growth factor. Recent Progr. Hormone Res. 30, 551–574 (1974)
Cooper, T.G., Danzo, B.J., DiPietro, D.L., McKenna, T.J., Orgebin-Crist, M.-C.: Some characteristics of rete testis fluid from rabbits. Andrologia (in press)
Danzo, B.J., Eller, B.C., Orgebin-Crist, M.-C.: Approaches to fertility regulation through an understanding of the molecular control of epididymal function. In: 5th International Seminar on Reproductive Physiology and Sexual Endocrinology: Sperm Action. pp. 74–88: Karger Basel 1976
Djøseland, O., Hansson, V., Haugen, H.N.: Androgen metabolism by rat epididymis. I. Metabolic conversion of 3H-testosterone in vivo. Steroids 21, 773–783 (1973)
Djøseland, O., Hanssen, V., Haugen, H.N.: Androgen metabolism by rat epididymis. II. Metabolic conversion of 3H-testosterone in vitro. Steroids 23, 397–410 (174)
Duncan, D.B.: Multiple range tests for correlated and heteroseedastic means. Biometrics 13, 164–176 (1957)
Dyson, A.L.M.B., Orgebin-Crist, M.-C.: Effect of hypophysectomy, castration and androgen replacement upon the fertilizing ability of rat epididymal spermatozoa. Endocrinology 93, 391–402 (1973)
Ewing, L., Brown, B., Irby, D.C., Jardine, I.: Testosterone and 5α-reduced androgen secretion by rabbit testes-epididymides perfused in vitro. Endocrinology 96, 610–617 (1975)
Farnsworth, W.E., Brown, J.R.: Metabolism of testosterone by the human prostate. J. Amer. med. Assoc. 183, 436–439 (1963)
Frankel, A.J., Eik-Nes, K.B.: Testosterone and dihydroepiandrosterone in the epididymis of the rabbit. J. Reprod. Fertil. 23, 441–445 (1970)
Gloyna, R.E., Wilson, J.D.: A comparative study of the conversion of testosterone to 17β-hydroxy5α-androstan-3-one (dihydrotestosterone) by prostate and epididymis. J. clin. Endocr. 29, 970–977 (1969)
Haltmeyer, G.C., Eik-Nes, K.B.: Plasma levels of testosterone in male rabbits following copulation. J. Reprod. 19, 273–277 (1969)
Hammond, J., Asdell, S.A.: The vitality of the spermatozoa in the male and female reproductive tracts. Brit. J. exp. Biol. 4, 155–185 (1926)
Hansson, V., Ritzén, E.M., French, F.S., Nayfeh, S.N.: Androgen transport and receptor mechanisms in testis and epididymis. In: Handbook of physiology (eds. D.W. Hamilton and R.O. Greep), sect. 7, vol. V, p. 143–172. Baltimore, Md.: Williams & Wilkins 1975
Hicks, J.J., Rojas, L., Rosado, A.: Insulin regulation of spermatozoa metabolism. Endocrinology 92, 833–839 (1973)
Ichihara, I, Santti, R.S., Pellerriemi, L.J.: Effect of testosterone, hydrocortisone, and insulin on the fine structure of the epithelium of rat ventral prostate in organ culture. Z. Zellforsch. 143, 425–438 (1973)
Inano, H., Machino, A., Tamaoki, B.: In vitro metabolism of steroid hormones by cell-free homogenates of epididymides of adult rats. Endocrinology 84, 997–1003 (1969)
Kotani, M., Seiki, K., Hattori, M.: Retardation of spermatogenesis and testosterone secretion after ligature of lymphatics draining the testes of rabbits. Endocr. jap. 21, 1–8 (1974)
Lasnitzki, I.: The action of hormones on cell and organ cultures. In: Cells and tissues in cultures; methods, biology and physiology (ed. E.N. Willmer), p. 591–658. New York: Academic Press 1965
Lasnitzki, I., Franklin, H.R.: The influence of serum on uptake, conversion, and action of testosterone in rat prostate glands in organ culture. J. Endocr. 54, 333–342 (1972)
Lasnitzki, I., Franklin, H.R.: The influence of serum on the uptake, conversion, and action of dihydrotestosterone in rat prostate glands in organ culture. J. Endocr. 64, 289–297 (1975)
Liao, S., Fang, S.: Receptor-proteins for androgens and the mode of action of androgens on gene transcription in ventral prostate. Vitam. and Horm. 27, 17–90 (1970)
Lostroh, A.J.: Effect of testosterone and insulin in vitro on maintenance and repair of the secretory epithelium of the mouse prostate. Endocrinology 88, 500–503 (1971)
Lubicz-Nawrocki, C.M.: The effect of metabolites of testosterone on the viability of hamster epididymal spermatozoa. J. Endocr. 58, 193–198 (1973)
Lubicz-Nawrocki, C.M., Glover, T.D.: The influence of the testis on the survival of spermatozoa in the epididymis of the golden hamster, Mesocricetus auratus. J. Reprod. Fertil. 34, 315–329 (1973)
Moor, B.C., Younglai, E.V.: Variations in peripheral levels of LH and testosterone in adult male rabbits. J. Reprod. Fertil. 42, 259–266 (1975)
Orgebin-Crist, M.-C.: Sperm maturation in rabbit epididymis. Nature (Lond.) 216, 816–818 (1967)
Orgebin-Crist, M.-C.: Maturation of spermatozoa in the rabbit epididymis: Fertilizing ability and embryonic mortality in does inseminated with epididymal spermatozoa. Ann. Biol. anim. 7, 373–389 (1967)
Orgebin-Crist, M.-C.: Maturation of spermatozoa in the rabbit epididymis: Effect of castration and testosterone replacement. J. exp. Zool. 185, 301–310 (1973)
Orgebin-Crist, M.-C., Tichenor, P.: Effect of testosterone on sperm maturation in vitro. Nature androgen replacement in the rabbit epididymis. Cell Tiss. Res. 148, 188–201 (1974)
Orgebin-Crist, M.-C., Tichenor, P.: A technique for studying sperm maturation in vitro. Nature (Lond.) 239, 227–228 (1972)
Orgebin-Crist, M.-C., Tichenor, P.: Effect of testosterone on sperm maturation in vivo. Nature (Lond.) 245, 328–329 (1973)
Robel, P., Lasnitzki, I., Baulieu, E.-E.: Hormone metabolism and action: Testosterone and metabolites in prostate organ culture. Biochimie 53, 81–96 (1971)
Rowe, P.H., Hopkinson, C.R.N., Shenton, J.C., Glover, T.D.: The secretion of LH and testosterone in the rabbit. Steroids 25, 313–321 (1975)
Santti, R.S., Johansson, R.: Some biochemical effects of insulin and steroid hormones on the rat prostate in organ culture. Exp. Cell Res. 77, 111–120 (1973)
Seiki, K., Kotani, M., Yamashita, A., Miyamoto, M., Horii, I.: Androgens in blood and lymph plasma from normal and castrated rabbits. J. Endocr. 42, 157–158 (1968)
Smith, O.W., Hafs, H.D.: Competitive protein binding and radioimmunoassay for testosterone in bulls and rabbits; blood serum testosterone after injection of LH or prolactin in rabbits. Proc. Soc. exp. Biol. (N.Y.) 142, 804–810 (1973)
Sowell, J.G., Eik-Nes, K.B.: Formation in vivo of 5α-dihydrotestosterone by the canine epididymis. Proc. Soc. exp. Biol. (N.Y.) 141, 827–829 (1972)
Steinberger, E., Steinberger, A., Perloff, W.H.: Initiation of spermatogenesis in vitro. Endocrinology 74, 788–792 (1964)
Temin, H.M., Pierson, R.W., Dulak, N.C.: The role of serum in the control of multiplication of avian and mammalian cells in culture. In: Growth, nutrition, and metabolism of cells in culture (eds. G.H. Rothblat and V.J. Cristofalo), vol. I, p. 50–81. New York: Academic Press 1972
Tesh, J.M., Glover, T.D.: Ageing of rabbit spermatozoa in the male tract and its effect on fertility. J. Reprod. Fertil. 20, 287–297 (1969)
Thorneycroft, I.H., Thorneycroft, N.K., Scaramuzzi, R.J., Blake, C.A.: Radioimmunoassay of serum LH and testosterone in male rabbits actively immunized against testosterone. Endocrinology 97, 301–306 (1975)
Tindall, D.J., French, F.S., Nayfeh, S.N.: Androgen uptake and binding in rat epididymal nuclei, in vivo. Biochem. biophys. Res. Commun. 49, 1391–1397 (1972)
Vreeburg, J.T.M., Scholte, H.R.: Localization of 5α-steroid reductase in the epididymis of the rat. Acta endocr. (Kbh.) Suppl. 177, 67 (1973)
Wilson, J.D.: Metabolism of testicular androgens. In: Handbook of physiology (eds. D.W. Hamilton and R.O. Greep,), sect. 7, vol. V, p. 491–508. Baltimore, Md.: Williams & Wilkins 1975
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Orgebin-Crist, MC., Jahad, N. & Hoffman, L.H. The effects of testosterone, 5α-dihydrotestosterone, 3α-androstanediol, and 3β-androstanediol on the maturation of rabbit epididymal spermatozoa in organ culture. Cell Tissue Res. 167, 515–525 (1976). https://doi.org/10.1007/BF00215181
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00215181