- Research article
- Open Access
The expression of inducible nitric oxide synthase (iNOS) in the testis and epididymis of rats with a dihydrotestosterone (DHT) deficiency
Cellular & Molecular Biology Letters volume 14, pages511–527 (2009)
In our previous studies, we showed that a finasteride-induced DHT deficiency may cause changes in the morphology of the seminiferous epithelium without any morphological alteration of the epididymis. In this study, we demonstrated the constitutive immunoexpression of inducible nitric oxide synthase (iNOS) in the testis and epididymis of Wistar rats treated with finasteride for 28 days (the duration of two cycles of the seminiferous epithelium) and 56 days (the duration of one spermatogenesis). We noted that a 56-day finasteride treatment mainly caused a decrease in the level of circulating DHT, as well as a statistically insignificant decrease in the level of T. The hormone deficiency also led to a change in the iNOS immnoexpression in the testis and epididymis of the finasteride-treated rats. In vitro, DHT did not modify NO production by the epithelial cells of the caput epididymis even when stimulated with LPS and IFNγ, but it did give rise to an increase in NO production by the epithelial cells of the cauda epididymis without the stimulation. DHT did not have a statistically significant influence on estradiol production by cultured, LPS- and IFNγ-stimulated epithelial cells from the caput and cauda epididymis. In conclusion, our data clearly indicates that a finasterideinduced DHT deficiency intensifies the constitutive expression of iNOS in most rat testicular and epididymal cells, so it can be expected that the expression of inducible nitric oxide synthase (iNOS) could be regulated by DHT. On the other hand, the profile of the circulating DHT and T levels strongly suggests that the regulation of constitutive iNOS expression is complex and needs more detailed study.
5α-reductase type 1
5α-reductase type 2
- E2 :
enzyme linked fluorescent assay
estrogen receptor α
estrogen receptor β
- NO2− :
nitric oxide synthase
endothelial nitric oxide synthase
inducible nitric oxide synthase
neuronal nitric oxide synthase
reverse transcriptase-polymerase chain reaction
Moncada, S., Palmer, R.M. and Higgs, E.A. Nitric oxide: physiology, phatophysiology, and pharmacology. Pharmacol. Rev. 43 (1991) 109–134.
Xie, Q. and Nathan, C. The high-output nitric oxide pathway: role and regulation. J. Leukoc. Biol. 56 (1994) 576–582.
Knowles, R.G. and Moncada, S. Nitric oxide synthases in mammals. Biochem. J. 298 (1994) 249–258.
Lowenstein, C.J., Glatt, C.S., Bredt, D.S. and Snyder, S.H. Cloned and expressed macrophage nitric oxide synthase contrasts with the brain enzyme. Proc. Natl. Acad. Sci. USA. 89 (1992) 6711–6715.
Kroncke, K.D., Fehsel, K. and Kolb-Bachofen, V. Inducible nitric oxide synthase and its product nitric oxide, a small molecule with complex biological activities. Biol. Chem. Hoppe. Seyler. 376 (1995) 327–343.
Burnett, A.L., Ricker, D.D., Chamness, S.L., Maguire, M.P., Crone, J.K., Bredt, D.S., Snyder, S.H. and Chang, T.S. Localization of nitric oxide synthase in the reproductive organs of the male rat. Biol. Reprod. 52 (1995) 1–7.
Ehren, I., Adolfsson, J. and Wiklund, N.P. Nitric oxide synthase activity in the human urogenital tract. Urol. Res. 22 (1994) 287–290.
Stephan, J.P., Guillemois, C., Jegou, B. and Bauche, F. Nitric oxide production by Sertoli cells in response to cytokines and lipopolysaccharide. Biochem. Biophys. Res. Commun. 213 (1995) 218–224.
Tatsumi, N., Fujisawa, M., Kanazaki, M., Okuda, Y., Okada, H., Arakawa, S. and Kamidono, S. Nitric oxide production by cultured rat Leydig cells. Endocrinology 138 (1997) 994–998.
Weissman, B.A., Niu, E., Ge, R., Sottas, C.M., Holmes, M., Hutson, J.C. and Hardy, M.P. Paracrine modulation of androgen synthesis in rat leydig cells by nitric oxide. J. Androl. 26 (2005) 369–378.
Wiszniewska, B., Kurzawa, R., Ciechanowicz, A. and Machaliński, B. Inducible nitric oxide synthase in the epithelial epididymal cells of the rat. Reprod. Fertil. Dev. 9 (1997) 789–794.
Kurzawa, R., Barcew-Wiszniewska, B. and Skowron, J. Rat epididymal epithelial cells produce nitric oxide upon concomitant lipopolysaccharide and interferon-gamma stimulation. Folia Histochem. Cytobiol. 4 (1996) 85–86.
O’Bryan, M.K., Schlatt, S., Gerdprasert, O., Phillips, D.J., de Kretser, D.M. and Hedger, M.P. Inducible nitric oxide synthase in the rat testis: evidence for potential roles in both normal function and inflammationmediated infertility. Biol. Reprod. 63 (2000) 1285–1293.
Ha, T.Y., Kim, H.S. and Shin, T. Expression of constitutive endothelial, neuronal and inducible nitric oxide synthase in the testis and epididymis of horse. J. Vet. Med. Sci. 66 (2004) 351–356.
Kim, H.C., Byun, J.S., Lee, T.K., Jeong, C.W., Ahn, M. and Shin, T. Expression of nitric oxide synthase isoform in the testes of pigs. Anat. Histol. Embryol. 36 (2007) 135–138.
Sun, L., Ren, Y.P., Jiang, W., Zhang, M.Y. and Hou, Q.Y. Expression and role of nitric oxide synthase in the testis and epididymis of Macaca fascicularis. Zhongua Nan Ke Xu 12 (2006) 876–878.
Jin, Y. and Penning, T.M. Steroid 5alpha-reductases and 3alphahydroxysteroid dehydrogenases: key enzymes in androgen metabolism. Best Pract. Res. Clin. Endocrinol. Metab. 15 (2001) 79–94.
Russell, D.W. and Wilson, J.D. Steroid 5α-reductase: two genes/two enzymes. Annu. Rev. Biochem. 63 (1994) 25–61.
Viger, R.S. and Robaire, B. Steady state steroid 5 alpha-reductase messenger ribonucleic acid levels and immunocytochemical localization of the type 1 protein in the rat testis during postnatal development. Endocrinology 136 (1995) 5409–5415.
Pratis, K., O’Donnell, L., Ooi, G.T., McLachlan, R.I. and Robertson, D.M. Enzyme assay for 5alpha-reductase type 2 activity in the presence of 5alpha-reductase type 1 activity in rat testis. J. Steroid. Biochem. Mol. Biol. 75 (2000) 75–82.
Viger, R.S. and Robaire, B. The mRNAs for the steroid 5α-reductase izoenzymes, types 1 and 2, are differentlly regulated in the rat epididymis. J. Androl. 17 (1996) 27–34.
Mahony, M.C., Swanlund, D.J., Billeter, M., Roberts, K.P. and Pryor, J.L. Regional distribution of 5alpha-reductase type 1 and type 2 mRNA along the human epididymis. Fertil. Steril. 69 (1998) 1116–1121.
Robaire, B. and Henderson, N.A. Actions of 5alpha-reductase inhibitors on the epididymis. Mol. Cell. Endocrinol. 250 (2006) 190–195.
Metcalf, B.W., Levy, M.A. and Holt, D.A. Inhibitors of steroid 5alphareductase in benign prostatic hyperplasia, male pattern baldness and acne. Trends Pharmacol. Sci. 10 (1989) 491–495.
George, F.W. Androgen metabolism in the prostate of the finasteridetreated, adult rat: a possible explanation for the differential action of testosterone and 5α-dihydrotestosterone during development of male urogenital tract. Endocrinology 138 (1997) 871–877.
Vaughan, E.D. Long-term experience with 5-alpha-reductase inhibitors. Rev. Urol. 5 (2003) S22–S27.
Kolasa, A., Marchlewicz, M., Wenda-Różewicka, L. and Wiszniewska, B. Morphology of the testis and the epididymis in rats with dihydrotestosterone (DHT) deficiency. Rocz. Akad. Med. Bialymst. 49 (2004) 117–119.
Wiszniewska, B. Steroidogenic characteristics of in vitro cultured epididymal epithelial cells of the rat. Reprod. Biol. 1 (2001) 60–66.
Ding, A.H., Nathan, C.F. and Stuehr, D.J. Release of reactive nitrogen intermediates and reactive oxygen intermediates from mouse peritoneal macrophages. Comparison of activating cytokines and evidence for independent production. J. Immunol. 141 (1988) 2407–2412.
Zini, A., Abitbol, J., Girardi, S.K., Schulsinger, D., Goldstein, M. and Schlegel, P.N. Germ cell apoptosis and endothelial nitric oxide synthase (eNOS) expression following ischemia-reperfusion injury to testis. Arch. Androl. 41 (1998) 57–65.
Lee, N.P. and Cheng, C.Y. Regulation of Sertoli cell tight junction dynamics in the rat testis via the nitric oxide synthase/soluble guanylate cyclase/3′,5′-cyclic guanosine monophosphate/protein kinase G signaling pathway: an in vitro study. Endocrinology 144 (2003) 3114–3129.
Lue, Y., Sinha-Hikim, A.P., Wang, C., Leung, A. and Swerdloff, R.S. Functional role of inducible nitric oxide synthase in the induction of male germ cell apoptosis, regulation of sperm number and determination of testis size: evidence from null mutant mice. Endocrinology 144 (2003) 3092–3100.
Zini, A., O’Bryan, M.K., Magid, M.S. and Schlegel, P.N. Immunohistochemical localization of endothelial nitric oxide synthase in human testis, epididymis, and vas deferens suggests a possible role for nitric oxide in spermatogenesis, germ maturation, and programmed cell death. Biol. Reprod. 55 (1996) 935–941.
Gerdprasert, O., O’Bryan, M.K., Muir, J.A., Caldwell, A.M., Schlatt, S., de Krester, D.M. and Hedger, M.P. The response of testicular leucocytes to lipopolysaccharide-induced inflammation: further evidence heterogeneity of the testicular macrophage population. Cell. Tissue Res. 308 (2002) 277–285.
Welsh, C., Watson, M.E., Poth, M., Hong, T. and Frrancis, G.L. Evidence to suggest nitric oxide is an interstitial regulator of Leydig Cell steroidogenesis. Metabolism 44 (1995) 234–238.
Wiszniewska, B. Primary culture of the rat epididymal epithelial cells as a source of oestrogen. Andrologia 34 (2002) 180–187.
Chamness, S.L., Ricker, D.D., Crone, J.K., Dembeck, C.L., Maguire, M.P., Burnett, A.L. and Chang, T.S. The effect of androgen on nitric oxide synthase in the male reproductive tract of the rat. Fertil. Steril. 63 (1995) 1101–1107.
Carreau S. The testicular aromatase: from gene to physiological role. Reprod. Biol. 2 (2002) 5–12.
Lambard, S., Galeraud-Denis, I., Saunders, P.T. and Carreau, S. Human immature germ cells and ejaculated spermatozoa contain aromatase and oestrogen receptors. J. Mol. Endocrinol. 32 (2004) 279–289.
Carpino, A., Romeo, F. and Rago, V. Aromatase immunolocalization in human ductuli efferentes and proximal ductus epididymis. J. Anat. 204 (2004) 217–220.
Shayu, D. and Rao, A.J. Expression of functional aromatase in the epididymis: role of androgens and LH in modulation of expression and activity. Mol. Cell. Endocrinol. 249 (2006) 40–50.
Snyder, G.D., Holmes, R.W., Bates, J.N. and Van Voorhis, B.J. Nitric oxide inhibits aromatase activity: mechanism of action. J. Steroid. Biochem. Mol. Biol. 58 (1996) 63–69.