Summary
The subcellular distribution of free and esterified cholesterol in mouse testis and the changes occurring in cholesterol content of whole testes and cell fractions after inhibition of gonadotropins with methallibure (ICI 33, 828) and restimulation with human chorionic gonadotropin (HCG) are reported in the present paper.
In subcellular fractions, the bulk of free cholesterol is associated with organelles sedimented in the microsomal fraction while esterified cholesterol is mainly stored in isolated lipid droplets. The latter compartment increases in methallibure-treated mice 2.3 fold and is remarkably depleted after administration of HCG. There is a close parallelism between the changes in esterified cholesterol content and the variation in the numbers of lipid droplets found in electron micrographs of interstitial cells of mice receiving similar treatments. By contrast no significant changes were noticed in either free cholesterol concentrations of the microsomal fractions or in the fine structure of organelles associated with this fraction. The dynamic nature of steroidogenesis in the microsomal fraction requires the existence of a free cholesterol pool with a high turnover rate for use as an intermediate in androgen synthesis. On the other hand, the large content of free cholesterol in microsomes and its stability under different conditions suggest the presence of a cholesterol compartment with a slow turnover, as a constituent of the membranes.
Similar content being viewed by others
References
Aoki, A.: Hormone-induced differentiation of agranular endoplasmic reticulum in the interstitial cells of the mouse testis. Protoplasma (Wien) 66, 263–267 (1968)
Aoki, A.: Hormonal control of Leydig cell differentiation. Protoplasma (Wien) 71, 209–225 (1970)
Aoki, A., Massa, E.M.: Early responses of testicular interstitial cells to stimulation by interstitial-cell-stimulating hormone. Amer. J. Anat. 134, 239–261 (1972)
Armstrong, D.T.: Gonadotropins, ovarian metabolism and steroid biosynthesis. Rec. Progr. Hormone Res. 24, 255–308 (1968)
Armstrong, D.T.: Reproduction. Ann. Rev. Physiol. 32, 439–470 (1970)
Bartke, A.: Concentration of free and esterified cholesterol in the testes of immature and adult mice. J. Reprod. Fertil. 25, 153–156 (1971a)
Bartke, A.: Effects of prolactin and luteinizing hormone on the cholesterol stores in the mouse testis. J. Endocr. 49, 317–323 (1971b)
Brown, P.S.: Observations on a dithiocarbamyolhydrazine as an inhibitor of pituitary gonadotrophic activity. J. Endocr. 26, 425–436 (1963)
Christensen, A.K.: The fine structure of testicular interstitial cells in guinea pigs. J. Cell Biol. 26, 911–935 (1965)
Christensen, A.K., Fawcett, D.W.: The normal fine structure of opossum testicular interstitial cells. J. biophysic. biochem. Cytol. 9, 653–670 (1961)
Christensen, A.K., Fawcett, D.W.: The fine structure of testicular interstitial cells in mice. Amer. J. Anat. 118, 551–572 (1966)
Connell, C.J.: The effect of luteinizing hormone on the ultrastructure of the Leydig cell of the chick. Z. Zellforsch. 128, 139–151 (1972)
Creech, E.G., Sewell, B.W.: A rapid micro procedure for determination of free and esterified cholesterol in blood serum using silicic acid separation. Analyt. Biochem. 3, 119–123 (1962)
Eik-Nes, K.B.: Production and secretion of testicular steroids. Rec. Progr. Hormone Res. 27, 517–535 (1971)
Enders, A.C., Lyons, W.R.: Observations on the fine structure of lutein cells. II. The effect of hypophysectomy and mammotrophic hormone in the rat. J. Cell Biol. 22, 127–141 (1964)
Fawcett, D.W.: Structural and functional variations in the membranes of the cytoplasm. In: Intracellular Membranous Structure. (S. Seno and E.V. Cowdry, eds.), p. 15–40. Okayama: Japan Society for Cell Biology 1965
Fawcett, D.W., Long, J.A., Jones, A.L.: The ultrastructure of endocrine glands. Rec. Progr. Hormone Res. 25, 315–368 (1969)
Folch, J., Lees, M., Sloane Stanley, G.H.: A simple method for the isolation and purification of total lipides from animal tissues. J. biol. Chem. 226, 497–509 (1957)
Goodman, D.S.: Cholesterol ester metabolism. Physiol. Rev. 45, 747–839 (1965)
Hall, P.F., Irby, C.C., de Kretser, D.M.: Conversion of cholesterol to androgens by rat testes: Comparison of interstitial cells and seminiferous tubules. Endocrinology 84, 488–496 (1969)
Hogeboom, G.H.: Fractionation of cell components of animal tissues. In: Methods in Enzymology. (S.P. Colowick and N.O. Kaplan, eds.), vol. I, p. 16–19. New York: Academic Press 1955
Horning, M.G., Williams, E.A., Horning, E.C.: Separation of tissue cholesterol esters and triglycerides by silicic acid chromatography. J. Lipid Res. 1, 482–485 (1960)
Ichihara, I.: Cholesterol changes in developing testicular interstitial cells of the mouse: Histochemical and biochemical study. Anat. Rec. 163, 595–602 (1969)
Long, J.A., Jones, A.L.: The fine structure of the zona glomerulosa and the zona fasciculata of the adrenal cortex of the opossum. Amer. J. Anat. 120, 463–471 (1967)
Massa, E.M.: Gonadotropic control of testicular cholesterol compartments and its correlation with the ultrastructure of Leydig cells. Ph. D. Dissertation. Facultad de Ciencias Quimicas, Universidad Nacional de Córdoba (1972)
Massa, E.M., Aoki, A.: Simple method for purification of glutaraldehyde. Acta physiol. lat.-amer. 21, 161–163 (1971)
Menon, K.M.J., Dorfman, R.I., Forchielli, E.: Influence of gonadotropin on the cholesterol-sidechain cleavage reaction by rat-testis mitochondrial preparations. Biochim. biophys. Acta (Amst.) 148, 486–494 (1967)
Mollenhauer, H.H.: Plastic embedding mixtures for use in electron microscopy. Stain Technol. 39, 111–114(1964)
Moses, H.L., David, W.W., Rosenthal, A.S., Garren, L.D.: Adrenal cholesterol: localization by electron-microscope autoradiography. Science 163, 1203–1205 (1969)
Moyle, W., Jungas, R., Creep, R.: Apparent need for cholesteryl palmitate hydrolysis before cholesterol sidechain cleavage in intact testicular tumor mitochondria. Fed. Proc. 29, 247 (abstract) (1970)
Palade, G.E., Siekevitz, P.: Liver microsomes. An integrated morphological and biochemical study. J. biophys. biochem. Cytol. 2, 171–198 (1956)
Parvinen, M., Hurne, P., Niemi, M.: Penetration of exogenous testosterone, pregnenolone, progesterone and cholesterol into seminiferous tubules of rat. Endocrinology 87, 1082–1084 (1970)
Pokel, J.D., Moyle, W.R., Greep, R.O.: Depletion of esterified cholesterol in mouse testes and Leydig cell tumors by luteinizing hormone. Endocrinology 91, 323–325 (1972)
Shikita, M., Tamaoki, B.: Testerone formation by subcellular particles of rat testes. Endocrinology 76, 563–569 (1965)
Tamaoki, B.: Steroidgenesis and cell structure. Biochemical pursuit of sites of steroid biosynthesis. J. Steroid Biochem. 4, 89–118 (1973)
Toren, D., Menon, K.M.J., Dorfman, R.I.: In vitro enzymatic cleavage of the cholesterol sidechain in rat testis preparations. Steroid 3, 381–390 (1964)
Venable, J.H., Coggeshall, R.: A simplified lead citrate stain for use in electron microscopy. J. Cell Biol. 25, 407–408 (1965)
Zlatkis, A., Zak, B.: Study of a new cholesterol reagent. Analyt. Biochem. 29, 143–148 (1969)
Author information
Authors and Affiliations
Additional information
Supported partially by grants from the CONICET, The Rockefeller Foundation, and The Ford Foundation.
Rights and permissions
About this article
Cite this article
Aoki, A., Massa, E.M. Subcellular compartmentation of free and esterified cholesterol in the interstitial cells of the mouse testis. Cell Tissue Res. 165, 49–62 (1975). https://doi.org/10.1007/BF00222799
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00222799