Summary
The testes of normal white adult rats were studied with histochemical techniques for oxidative enzymes. Smears, imprints and squash preparations were used to obtain isolated cells.
LDH, HDH and NADH2-diaphorase rich granules were found close to the Golgi apparatus of spermatocytes.
An increase in some enzymes was described in spermatids during the formation process of the mitochondrial helix. The differences in enzymatic properties of the mitochondrial helix and those of the cytoplasmic mitochondria suggest a process of “selection” and differentiation.
Using a LDH-PMS medium, we found three loci of enzymatic activity, a diffuse cytoplasmic one, a Golgian activity, and a mitochondrial one. Applying Brody's method for the differentiation of LDH isozymes we demonstrated a preponderance of an isozyme non-sensible to urea or excess of lactate. This probably reflected the existence of the X-isozyme in the germinal line.
All tested enzymes except MAO were positive in spermatozoa. The great activity of GDH and HDH in the middle piece provides a good link between the energetic metabolism of the spermatozoid and the oxidation of lipids.
The enzymes of the Krebs' cycle, SDH, MDH and IDH were all present and we also demonstrated the presence of CO in the midpiece.
G-6-PDH was present in moderate amounts.
Great concentrations of these enzymes were found in Sertoli cells. Any conclusion about enzymatic distribution in sections must consider this fact.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ambadkar, P. M., George, J. C.: Histochemical localization of certain oxidative enzymes in the rat testis. J. Histochem. Cytochem. 12, 587–590 (1964).
Avers, C. J., Tkal, M. M.: Intracellular mitochondrial variation in enzyme activity as shown by histochemical studies using light and electron microscopy. J. Histochem. Cytochem. 11, 157–162 (1963).
Barka, T., Anderson, P. J.: Histochemistry. Theory, practice and bibliography. New York: Hoeber Medical Division, Harper and Row 1963.
Bishop, D. W.: Sperm motility. Physiol. Rev. 42, 1–59 (1962).
Blanco, A., Zinkham, W. H.: Lactate dehydrogenases in human testes. Science 139, 601–602 (1963).
Brody, I. A.: Isozyme histochemistry: A new method for the display of selective lactate dehydrogenase isozymes on an electrophoretic pattern. Nature (Lond.) 201, 685–687 (1964).
Burstone, M. S.: New histochemical technique for the demonstration of tissue oxidase (Cytochrome oxidase). J. Histochem. Cytochem. 7, 112–122 (1959).
Buta, J. L., Conklin, J. L., Dewey, M. M.: Subfractions of lactate dehydrogenase of the rat. J. Histochem. Cytochem. 14, 658–662 (1966).
Clermont, Y.: Cinétique de la spermatogenèse chez les mammifères. Arch. Anat. micr. 56, Suppl., 7–60 (1967).
Darlington, C. D., La Cour, L. F.: The handling of chromosomes. London: Allen & Unwin 1947.
Fahimi, H. D., Amarasingham, C. R.: Cytochemical localization of lactic dehydrogenase in white skeletal muscle. J. Cell Biol. 22, 29–48 (1964).
Farriaux, J. P., Fontaine, G.: Étude des isozymes de la L. D. H. dans le testicule et le sperme humains. Ann. Biol. clin. 25, 815–823 (1967).
Gerebtzoff, M. A.: Détection histochimique d'isoenzymes de la lactate deshydrogénase dans le nerf et le ganglion spinal. C. R. Soc. Biol. (Paris) 160, 1323–1325 (1966).
Glenner, G. G., Burtner, H. J., Brown, G. W.: The histochemical demonstration of monoamine-oxidase activity by tetrazolium salts. J. Histochem. Cytochem. 5, 591–600 (1957).
Goldberg, E., Hawtrey, C.: The ontogeny of sperm specific lactate dehydrogenase in mice. J. exp. Zool. 164, 309–316 (1967).
Goldfischer, S., Villaverde, H., Forschirm, R.: The demonstration of acid hydrolase, thermostable reduced diphosphopyridine nucleotide tetrazolium reductase and peroxidase activities in human lipofuscin pigment granules. J. Histochem. Cytochem. 14, 641–652 (1966).
Hashimoto, T., Kaluza, J. S., Burstone, M. S.: The effect of menadione and phenazine methosulfate on the tetrazolium reduction system under histochemical conditions. J. Histochem. Cytochem. 12, 797–804 (1964).
Holt, S. J., Hicks, R. M.: Studies on formalin fixation for electron microscopy and cytochemical staining purposes. J. biophys. biochem. Cytol. 11, 31–45 (1961).
Lardy, H. A., Hansen, R. G., Phillips, P. H.: The metabolism of bovine epididymal spermatozoa. Arch. Biochem. 6, 41–51 (1945).
Likar, I. N., Rosenkrantz, H.: Succinic dehydrogenase and Golgi complex of the grandular epithelia in the ventral prostatic gland of the sexually mature albino rat. Lab. Invest. 13, 246–249 (1964).
Mohri, H., Mohri, T., Ernster, L.: Isolation and enzymic properties of the midpiece of bull spermatozoa. Exp. Cell Res. 38, 217–246 (1965).
Nachlas, M. M., Tsou, K. C., De Souza, E., Chen, C. S., Seligman, A. M.: Cytochemical demonstration of succinic dehydrogenase by the use of a new p-nitrophenyl substituted ditetrazole. J. Histochem. Cytochem. 5, 420–436 (1957).
—, Walker, D. G., Seligman, A. M.: A histochemical method for the demonstration of diphosphopyridine nucleotide diaphorase. J. biophys. biochem. Cytol. 4, 29–38 (1958).
Niemi, M., Ikonen, M.: Cytochemistry of oxidative enzyme system in the Leydig cells of the rat testis and their functional significance. Endocrinology 70, 167–174 (1962).
Pearse, A. G. E.: Histochemistry. Theoretical and applied. London: Churchill Ltd. 1960.
Posalaki, Z., Szabó, D., Bácsi, E., Ökrös, I.: Hydrolytic enzymes during spermatogenesis in rat. An electron microscopic and histochemical studies. J. Histochem. Cytochem. 16, 249–262 (1968).
Reissenweber, N. J.: Succinic dehydrogenase activity in rat spermatozoa as demonstrated with various tetrazolium salts. Naturwissenschaften 53, 133–134 (1966).
Spector, R. G., Mutton, D. E., Hamerton, J. L.: Intracellular localization of four dehydrogenases in human skin fibroblasts in tissue culture. Nature (Lond.) 202, 672–674 (1964).
Turpeinen, P., Turpeinen, O., Talanti, S.: Effect of local heat in vivo on hyaluronidase, succinic dehydrogenase and phosphatases of the rat testis. Endocrinology 70, 731–737 (1962).
Walker, D. G., Seligman, A. M.: Formalin fixation in the cytochemical demonstration of succinic dehydrogenase of mitochondria. J. biophys. biochem. Cytol. 9, 415–427 (1961).
—: The use of formalin fixation in the cytochemical demonstration of succinic and DPN- and TPN-dependent dehydrogenases in mitochondria. J. Cell Biol. 16, 455–469 (1963).
White, I. G.: Metabolism of glycerol and similar compounds by bull spermatozoa. Amer. J. Physiol. 189, 307–310 (1957).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Reissenweber, N.J., Arbes-Navarrete, I. & Sala, M.A. Histochemical study of isolated intratubular cells of the rat testis. Histochemie 21, 84–96 (1970). https://doi.org/10.1007/BF00304808
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00304808