Summary
An improved method for the demonstration of 3β and 17β hydroxysteroid dehydrogenase (HSD) in human testis has been devised. The procedure maintains good cellular morphology with little loss of enzymatic activity. The significant features of the procedure are as follows:
-
1.
Tris-maleate buffer made up in Tyrode's balanced salt solution is employed in all steps from pre-freezing infiltration through fixation.
-
2.
Dimethylsulfoxide is used to protect from freezing artifacts, to enhance substrate and tetrazolium salt penetration, and to increase substrate solubility.
-
3.
A pre-incubation soak in the medium buffer system, dimethyl-sulfoxide, polyvinyl pyrrolidone and dimethylformamide, is used and a pH of 7.4 to 7.5 is maintained to limit false positive reaction.
-
4.
Potassium cyanide is employed in the medium to enhance the formation of reaction product.
-
5.
Polyvinylpyrrolidone is used in the pre-incubation and incubation media to reduce destruction of tissue morphology.
-
6.
Several steroid substrates for both 3β and 17β-HSD may be used.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Agger, P.: Scrotal and testicular temperature: Its relation to sperm count before and after operation for varicocele. Fertil. and Steril. 22, 286–297 (1971).
Allen, J. M.: A histochemical study of 3(β)-sterol dehydrogenase in the adrenal cortex of the developing mouse. Anat. Rec. 137, 57–63 (1960).
Baillie, A. H., Mack, W. S.: Hydroxysteroid dehydrogenase in normal and abnormal human testes. J. Endocr. 35, 239–248 (1966).
Baillie, A. H., Niemi, M., Ikonen, M.: 3β-hydroxysteroid dehydrogenase activity in the human foetal testis. Acta endocr. (Kbh.) 48, 429–438 (1965).
Bell, J. B. G.: Interconversion of testosterone and androstenedione in the human testis: a comparison between the activities displayed by the interstitium and the seminiferous tubules. Experientia (Basel) 28, 212–213 (1972).
Christensen, A. K., Mason, N. R.: Comparative ability of seminiferous tubules and interstitial tissue of rat testes to synthesize androgens from progesterone-4-14C in vitro. Endocrinology 76, 646–656 (1965).
Deane, H. W., Rubin, B. L.: Identification and control of cells that synthesize steroid hormones in the adrenal glands, gonads and placentae of various mammalian species. Arch. Anat. micr. 54, 49–66 (1965).
Fabbrini, A., Spera, G., Re, M., Savioli, M.: Histochemical techniques in the study of steroid dehydrogenases in human testis. Folia endocr. (Roma) 28, 533–543 (1970).
Goldberg, B., Jones, G. E. S., Borkowf, H. I.: A histochemical study of substrate specificity for the steroid 3β-ol dehydrogenase and isomerase systems in human ovary and testis. J. Histochem. Cytochem. 12, 880–889 (1964).
Hall, P. F., Irby, D. C., Kretser, D. M. de: Conversion of cholesterol to androgens by rat testes: comparison of interstitial cells and seminiferous tubules. Endocrinology 84, 488–496 (1969).
Jirásek, J. E., Raboch, J.: Histochemical study of testicular material from patients with various disorders. Fertil. and Steril. 14, 237–245 (1963).
Jones, G. E. S., Goldberg, B., Woodruff, J. D.: Cell specific steroid inhibitions in histochemical steroid 3β-ol dehydrogenase activities in man. Histochemie 14, 131–142 (1968).
Koudstaal, J., Frensdorf, E. L., Kremer, J., Mudde, J. M., Hardonk, M. J.: The histochemical pattern of the human adult testes. Acta endocr. (Kbh.) 55, 415–426 (1967).
Levy, H., Deane, H. W., Rubin, B. L.: Visualization of steroid-3β-ol dehydrogenase activity in tissues of intact and hypophysectomized rats. Endocrinology 65, 932–943 (1959).
Maeir, D. M.: Species variation in testicular Δ5-3β-hydroxysteroid dehydrogenase activity: Absence of activity in primate Leydig cells. Endocrinology 76, 463–469 (1965).
Magrini, V., Bertoli, G., Mazzola, R.: Histochemistry of 3β-hydroxysteroid dehydrogenase in human testis in normal, dysgenetic and various pathological conditions. Path. europ. 4, 18–29 (1969).
Mancini, R. E., Vilar, O., Lavieri, J. C., Andrada, J. A., Heinrich, J. J.: Development of Leydig cells in the normal human testis: A cytological, cytochemical and quantitative study. Amer. J. Anat. 112, 203–214 (1963).
Pearson, B., Grose, F.: Histochemical demonstration of 17β-Hydroxysteroid dehydrogenase by use of tetrazolium salt. Proc. Soc. exptl. Biol. (N.Y.) 100, 636–638 (1959).
Piotti, L. E., Ghiringhelli, F., Magrini, V.: A propos de la fonction testiculaire du vieillard: observations histochemiques et biologiques. Rev. franc. Endocr. clin. 8, 479–491 (1967).
Rowley, M. J., Heller, C. G.: The testicular biopsy: surgical procedure, fixation, and staining technics. Fertil. and Steril. 17, 177–186 (1966).
Rubin, B. L., Deane, H. W., Hamilton, J. A., Driks, E. C.: Changes in Δ 5-3β-Hydroxysteroid dehydrogenase activity in the ovaries of maturing rats. Endocrinology 72, 924–930 (1963).
Sengoku, K.: Histochemical studies on the human testes, especially on the interstitial cells of Leydig. Bull. Tokyo Med. Dent. Univ. 14, 51–78 (1967).
Sniffen, R. C.: The testis. I. The normal testis. Arch. Path. 50, 259–284 (1950).
Wattenberg, L. W.: Microscopic histochemical demonstration of steroid-3β-ol dehydrogenase in tissue sections. J. Histochem. Cytochem. 6, 225–232 (1958).
Zimmermann, H., Pearse, A. G. E.: Limitations in the histochemical demonstration of pyridine nucleotide linked dehydrogenase (“nothing dehydrogenase”). J. Histochem. Cytochem. 7, 271–275 (1959).
Author information
Authors and Affiliations
Additional information
Supported by Grant No. 68-806 from The Ford Foundation and Contract No. AT (45-1) 1780 from the U.S. Atomic Energy Commission.
Rights and permissions
About this article
Cite this article
Morse, H.C., Heller, C.G. Improvements in technique for the histochemical demonstration of 3β and 17β hydroxysteroid dehydrogenase in human testis. Histochemie 35, 331–339 (1973). https://doi.org/10.1007/BF00310672
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00310672