Molecular and Cellular Biochemistry

, Volume 292, Issue 1–2, pp 27–38 | Cite as

Role of selenium in spermatogenesis: Differential expression of cjun and cfos in tubular cells of mice testis

  • Sonia Shalini
  • Mohinder P. Bansal


Selenium (Se) is an essential dietary trace element, involved in the process of male reproduction. Best known as an antioxidant, it acts through various selenoproteins viz. glutathione peroxidase, thioredoxin reductase and selenoprotein P. The aim of the present study was to identify the underlying molecular mechanism of Se in regulating spermatogenesis. Different Se status: deficient, adequate and excess Se, were generated in male Balb/c mice by feeding yeast based Se deficient diet, and deficient diet supplemented with Se as sodium selenite (0.2 and 1 ppm Se) respectively for a period of 4 and 8 weeks. Se levels and glutathione peroxidase (GSH-Px) activity were significantly reduced in the Se deficient mice and enhanced in Se supplemented group. Reduction in the number of post-meiotic germ cells viz. spermatids and spermatozoa, were observed in the deficient groups indicating loss in fertility and reproductive ability. cjun and cfos (components of transcription factor AP1) regulate cellular growth and differentiation and also exert a regulatory role in steroidogenesis and spermatogenesis. Changes in the mRNA expression of cjun and cfos were observed. Concomitant with this, western blot revealed that the protein expression profile for both these genes was significantly altered in the Se deficient and Se excess groups. Further immunohistochemical analysis showed that, both these genes had identical cellular localization indicating that they do not work alone but act synergistically as AP1. cjun and cfos expression was greater in the early mitotic stages-spermatogonia and spermatocytes in the Se adequate controls. It decreased in the meiotic stages and then again peaked around the later stages-elongating spermatids and spermatozoa. However in the Se deficient mice, weaker expression was observed in the spermatogonia with a complete absence of expression near the lumen. No visible changes in cjun/cfos expression and immunohistochemical localization were observed in the excess group compared to the Se adequate controls. In conclusion, the present study clearly demonstrates that alteration in Se supply leads to decreased expression pattern for both cJun and cFos in the testicular germ cells which might be responsible for decreased germ cell number, differentiation and reduced fertility and accounts for the mechanism of Se action in regulating spermatogenesis.


testes cjun cfos western blot immunohistochemistry selenium 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Behne D, Duk M, Elger W: Selenium content and glutathione peroxidase activity in the testis of the maturing rat. J Nutr 116: 1442–1447, 1986PubMedGoogle Scholar
  2. 2.
    Oldereid NB, Thomassen Y, Purvis K: Selenium in human male reproductive organs. Hum. Reprod 13: 2172–2176, 1998PubMedCrossRefGoogle Scholar
  3. 3.
    Sprinker LH, Harr JR, Newberne PM, Whanger PD, Weswig PH: Selenium deficiency lesions in rats fed vitamin E supplemented rations. Nutr Reports Int 4: 335–339, 1971Google Scholar
  4. 4.
    Wallace E, Calvin HI, Plotz K, Cooper GW: Functional and developmental studies on the role of selenium in spermatogenesis. In: G.F. Combs, O.A. Levander, J.E. Spallholz, J.E. Oldfield (eds), Selenium biology and medicine Vol A. New York: AVI; pp. 181–196, 1987Google Scholar
  5. 5.
    Watanbe T, Endo A: Effects of selenium deficiency on sperm morphology and spermatocyte chromosomes in mice. Mutat Res 262: 93–99, 1991CrossRefGoogle Scholar
  6. 6.
    Behne D, Weiler H, Kyriakopoulos A: Effects of Selenium deficiency on testicular morphology and functions in rats. J Reprod Fertil 106: 291–297, 1996PubMedCrossRefGoogle Scholar
  7. 7.
    Bleau G, Lemarbre J, Faucher G, Roberts KD, Chapdelaine A: Semen selenium and human fertility. Fert Steril 42: 890–894, 1984Google Scholar
  8. 8.
    Kaur R, Prashad V: Effects of dietary selenium on differentiation, morphology and functions of spermatozoa of the house mouse, rat, Rattus-Rattus L. Mutat Res. Fundamental and molecular mechanisms of mutagenesis 309: 29–35, 1994CrossRefGoogle Scholar
  9. 9.
    Wolfes H, Kogawa K, Millette CF, Cooper GM: Specific expression of nuclear proto-oncogenes before entry into meiotic prophase of spermatogenesis. Science 245: 740–743, 1989PubMedCrossRefGoogle Scholar
  10. 10.
    Sorrentino V, McKinney MD, Giorgi M, Geremia R, Fleissner E: Expression of Cellular Protooncogenes in the Mouse Male Germ Line: A Distinctive 2.4-Kilobase pim-1 Transcript is Expressed in Haploid Postmeiotic Cells. PNAS 85: 2191–2195, 1988PubMedCrossRefGoogle Scholar
  11. 11.
    Cohen DR, Curran T: Analysis of dimerization and DNA binding functions in Fos and Jun by domain-swapping: involvement of residues outside the leucine zipper/basic region. Oncogene 5(6): 929–939, 1990PubMedGoogle Scholar
  12. 12.
    Wilkinson DG, Bhatt S, Ryseck RP, Bravo R: Tissue-specific expression of c-jun and junB during organogenesis in the mouse. Development 106: 465–71, 1989PubMedGoogle Scholar
  13. 13.
    Alcivar AA, Hake LE, Hardy MP, Hecht NB: Increased levels of junB and c-jun mRNAs in male germ cells following testicular cell dissociation. Maximal stimulation in prepuberal animals. J Biol Chem 265: 20160–165, 1990PubMedGoogle Scholar
  14. 14.
    Kierszenbaum AL: Mammalian spermatogenesis in vivo and in vitro: a partnership of spermatogenic and somatic cell lineages. Endocr Rev 15: 116–34, 1994PubMedCrossRefGoogle Scholar
  15. 15.
    Cohen DR, Vandermark SE, McGovern JD, Bradley MP: Transcriptional regulation in the testis: a role for transcription factor AP-1 complexes at various stages of spermatogenesis. Oncogene 8(2): 443–455, 1993PubMedGoogle Scholar
  16. 16.
    Cobellis G, Rosaria M, Giulia F, Riccardo P, Silvia F: Cytoplasmic and nuclear fos protein forms regulate resumption of spermatogenesis in the frog, Rana esculenta. Endocrinology 143: 163–170, 2002PubMedCrossRefGoogle Scholar
  17. 17.
    Abate C, Patel L, Rausher FJ, Curran T: Redox regulation of Fos and Jun DNA-binding activity in vitro. Science 249: 1157–1161, 1990PubMedCrossRefGoogle Scholar
  18. 18.
    Burk RF: Production of selenium deficiency in the rat. Methods Enzymol 143: 307–313, 1987PubMedCrossRefGoogle Scholar
  19. 19.
    Hasunuma R, Ogawi T, Kawaniska Y: Fluorometric determination of selenium in nanogram amounts in biological materials using 2,3-diaminonaphthalene. Anal Biochem 126: 242–245, 1982PubMedCrossRefGoogle Scholar
  20. 20.
    Lawrence RA, Burk RF: Glutathione peroxidase activity in Selenium deficient rat liver. Biochem Biophy Res Commun 71: 952–958, 1976CrossRefGoogle Scholar
  21. 21.
    Wills ED: Mechanisms of lipid peroxide formation in animal tissues. Biochem J 99(3): 6676–6676, 1966Google Scholar
  22. 22.
    LeBlond CP, Clermont Y: Defination of the stages of the cycle of the seminiferous epithelium in the rat. Ann NY Acad Sci. 55: 548–573, 1952PubMedGoogle Scholar
  23. 23.
    Dignam JD, Lebovitz RM, Roeder RG: Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res 11: 1475–1489, 1983PubMedGoogle Scholar
  24. 24.
    Lowry OH, Rosebrough NJ, Farr AL, Randell RJ: Protein measurement with folin-phenol reagent. J Biol Chem 193: 265–275, 1951PubMedGoogle Scholar
  25. 25.
    Hafeman DG, Sunde RA, Hoekstra WG: Effect of dietary selenium on erythrocyte and liver glutathione peroxidase in rat. J Nutr 104: 580–587, 1974PubMedGoogle Scholar
  26. 26.
    Griveau JF, Dumont E, Renard P, Callegari JP, Le Lannou D: Reactive oxygen species, lipid peroxidation and enzymatic defence systems in human spermatozoa. J Reprod Fert 103: 17–26, 1995CrossRefGoogle Scholar
  27. 27.
    Surai P, Kostjuk I, Wishart G, Macpherson A, Speake B, Noble R, Ionov I, Kutz E: Effect of vitamin E and selenium supplementation of cockerel diets on glutathione peroxidase activity and lipid peroxidation susceptibility in sperm, testes, and liver. Biol Trace Elem Res 64(1–3): 119–132, 1998PubMedGoogle Scholar
  28. 28.
    Lane HW, Medina D: Mode of action of selenium inhibition of 7,12-dimethylbenz[a]anthracene-induced mouse mammary tumorigenesis. J Natl Cancer Inst 75(4): 675–679, 1985PubMedGoogle Scholar
  29. 29.
    Shen H-M, Yang C-F, Liu J, Ong C-N: The pro- and antioxidant role of glutathione in selenite-induced oxidative stress and apoptosis. Ann NY Acad Sci928: 355, 2001CrossRefGoogle Scholar
  30. 30.
    Aitken RJ: Free radicals, lipid peroxidation, and sperm function. Reprod Fertil Dev 7: 659–668, 1995PubMedCrossRefGoogle Scholar
  31. 31.
    Chen CS, Chao HT, Pan RL, Wei YH: Hydroxyl radical – induced decline in motility and increase in lipid peroxidation and DNA modification in human sperm. Biochem Mol Biol Int 43: 291–303, 1997PubMedGoogle Scholar
  32. 32.
    Kaur P, Bansal MP: Effect of selenium induced oxidative stress on the oxidation-reduction ability of male mice. Biol Trace Ele Res 97: 83–94, 2004CrossRefGoogle Scholar
  33. 33.
    Park HS, Park E, Kim MS, Ahn K, Kim IY, Choi EJ: Selenite inhibits the c-Jun N-terminal Kinase/Stress-activated Protein Kinase (JNK/SAPK) through a Thiol Redox Mechanism. J Biol Chem 275: 2527–2531, 2000PubMedCrossRefGoogle Scholar
  34. 34.
    Schultz R, Penttila TL, Parvinen M, Persson H, Hokfelt T, Pelto-Huikko M: Expression of immediate early genes in tubular cells of rat testis. Biol Reprod 52: 1215–1226, 1995PubMedCrossRefGoogle Scholar
  35. 35.
    Angel P, Karin M: The role of Jun, Fos and the AP-1 complex in cell-proliferation and transformation. Biochim Biophy Acta 1072: 129–157, 1991Google Scholar
  36. 36.
    Thepot D, Weitzman JB, Barra J, Segretain D, Stinnakre MG, Babinet C, Yaniv M: Targeted disruption of the murine junD gene results in multiple defects in male reproductive function. Development 127: 143–153, 2000PubMedGoogle Scholar
  37. 37.
    Yuan S, Xu S, Yang X, Liu X, Hao J, Qian M: Effects of c-jun on hCG-induced testosterone secretion of rat Leydig cells in vitro. Zhonghua Nan Ke Xue: 10(5) 345–7, 2004PubMedGoogle Scholar
  38. 38.
    Handel ML, Watts CKW, DeFazio A, Day RO, Sutherland RL: Inhibition of AP-1 binding and transcription by gold and selenium involving conserved cysteine residues in Jun and Fos. Proc Natl Acad Sci USA 92: 4497–4501, 1995Google Scholar
  39. 39.
    Chieffi P: Changes in JNK1 activity in the frog (Rana esculenta) testis. Mol Reprod Dev: 66(4) 398–402, 2003PubMedCrossRefGoogle Scholar
  40. 40.
    Pelto-Huikko M, Schultz R, Koistinaho J, Hokfelt T: Immunocytochemical demonstration of c-fos protein in sertoli cells and germ cells in rat testis. Acta Physiol Scand 141(2): 283–284, 1991PubMedCrossRefGoogle Scholar
  41. 41.
    Grisworld, MD, Kim JS, Tribley WA: Mechanisms involved in the homologous down-regulation of transcription of the follicle-stimulating hormone receptor gene in Sertoli cells. Mol Cell Endocrinol 173(1–2): 95–107, 2001CrossRefGoogle Scholar
  42. 42.
    Yamada K, Takane-Gyotoku N, Yuan X, Ichikawa F, Inada C, Nonaka K: Mouse islet cell lysis mediated by interleukin-1-induced Fas. Diabetologia 39: 1306–1312, 1996PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  1. 1.Department of BiophysicsPanjab UniversityChandigarhIndia

Personalised recommendations