Skip to main content
SpringerLink
Log in

The Effects of Di(2-Ethylhexyl)Phthalate Exposure and Selenium Nutrition on Sertoli Cell Vimentin Structure and Germ-Cell Apoptosis in Rat Testis

  • Published:
Archives of Environmental Contamination and Toxicology Aims and scope Submit manuscript

Abstract

This study aimed to investigate the effects of di(2-ethylhexyl)phthalate (DEHP) on Sertoli-cell vimentin filaments and germ-cell apoptosis in testes of pubertal rats at different selenium (Se) status. Se deficiency was produced in 3-weeks old Sprague-Dawley rats by feeding them ≤0.05 Se mg/kg diet for 5 weeks, Se supplementation group was on 1 mg Se/kg diet, and DEHP was applied at 1000 mg/kg dose by gavage during the last 10 days of the feeding period. The diet with excess Se did not cause any appreciable alteration in vimentin staining and apoptosis of germ cells, but Se deficiency caused a mild decrease in the intensity of vimentin immunoreactivity and enhanced germ-cell apoptosis significantly (approximately 3-fold, p <0.0033). DEHP exposure caused disruption and collapse of vimentin filaments and significantly induced apoptotic death of germ cells (approximately 8-fold, p <0.0033). In DEHP-exposed Se-deficient animals, compared with the control, collapse of vimentin filaments was more prominent; there was serious damage to the seminiferous epithelium; and a high increment (approximately 25-fold, p <0.0033) in apoptotic germ cells was observed. Thus, Se deficiency exacerbated the toxicity of DEHP on Sertoli cells and spermatogenesis, whereas Se supplementation provided protection. These results put forward the critical role of Se in the modulation of redox status of testicular cells and emphasize the importance of Se status for reproductive health.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Aitken RJ, Baker MA (2006) Oxidative stress, sperm survival and fertility control. Mol Cell Endocrinol 250:66–69

    Article  CAS  Google Scholar 

  • Alam MS, Ohsako S, Tay TW, Tsunekawa N, Kanai Y, Kurohmaru M (2010) Di(n-butyl)phthalate induces vimentin filaments disruption in rat Sertoli cells: a possible relation with spermatogenic cell apoptosis. Anat Histol Embryol 39:186–193

    Article  CAS  Google Scholar 

  • Allard EK, Johnson KJ, Boekelheide K (1993) Colchicine disrupts the cytoskeleton of rat testis seminiferous epithelium in a stage-dependent manner. Biol Reprod 48:143–153

    Article  CAS  Google Scholar 

  • Amlani S, Vogl AW (1988) Changes in the distribution of microtubules and intermediate filaments in mammalian Sertoli cells during spermatogenesis. Anat Rec 220:143–160

    Article  CAS  Google Scholar 

  • Bardin CW, Cheng CY, Musto NA, Gunsalus GL (1988) The Sertoli cell. In: Knobil E, Neill JD, Ewing LL, Greenwald GS, Markert CL, Pfaff DW (eds) The physiology of reproduction. Raven Press, New York, NY, pp 933–974

    Google Scholar 

  • Barlow NJ, Phillips SL, Wallace DG, Sar M, Gaido KW, Foster PM (2003) Quantitative changes in gene expression in fetal rat testes following exposure to di(n-butyl)phthalate. Toxicol Sci 7:431–441

    Article  Google Scholar 

  • Behne D, Höfer T, von Berswordt-Wallrabe R, Elger W (1982) Selenium in the testis of the rat: studies on its regulation and its importance for the organism. J Nutr 112:1682–1687

    CAS  Google Scholar 

  • Blount BC, Silva MJ, Caudill SP, Needham LL, Pirkle JL, Sampson EJ et al (2000) Levels of seven urinary phthalate metabolites in a human reference population. Environ Health Perspect 108:979–982

    Article  CAS  Google Scholar 

  • Borch J, Metzdorff SB, Vinggaard AM, Brokken L, Dalgaard M (2006) Mechanisms underlying the anti-androgenic effects of diethylhexyl phthalate in fetal rat testis. Toxicology 223:144–155

    Article  CAS  Google Scholar 

  • Dalgaard M, Nellemann C, Lam HR, Sørensen IK, Ladefoged O (2001) The acute effects of mono(2-ethylhexyl)phthalate (MEHP) on testes of prepubertal Wistar rats. Toxicol Lett 122:69–79

    Article  CAS  Google Scholar 

  • Diemer T, Allen JA, Hales KH, Hales DB (2003) Reactive oxygen disrupts mitochondria in MA-10 tumor Leydig cells and inhibits steroidogenic acute regulatory (StAR) protein and steroidogenesis. Endocrinology 144:2882–2891

    Article  CAS  Google Scholar 

  • Dröge W (2002) Free radicals in the physiological control of cell function. Physiol Rev 82:47–95

    Google Scholar 

  • Durmaz E, Ozmert EN, Erkekoglu P, Giray B, Derman O, Hincal F et al (2010) Plasma phthalate levels in pubertal gynecomastia. Pediatrics 125:e122–e129

    Article  Google Scholar 

  • Erkekoglu P, Rachidi W, De Rosa V, Giray B, Favier A, Hincal F (2010a) Protective effect of selenium supplementation on the genotoxicity of di(2-ethylhexyl)phthalate and mono(2-ethylhexyl)phthalate treatment in LNCaP cells. Free Radic Biol Med 49:559–566

    Article  CAS  Google Scholar 

  • Erkekoglu P, Rachidi W, Yuzugullu OG, Giray B, Favier A, Ozturk M, Hincal F (2010b) Evaluation of cytotoxicity and oxidative DNA damaging effects of di(2-ethylhexyl)-phthalate (DEHP) and mono(2-ethylhexyl)phthalate (MEHP) on MA-10 Leydig cells and protection by selenium. Toxicol Appl Pharmacol 248:52–62

    Article  CAS  Google Scholar 

  • Erkekoglu P, Zeybek ND, Giray B, Asan E, Arnaud J, Hincal F (2011a) Reproductive toxicity of di(2-ethylhexyl)phthalate in selenium-supplemented and selenium-deficient rats. Drug Chem Toxicol 34:379–389

    Article  CAS  Google Scholar 

  • Erkekoglu P, Rachidi W, Yüzügüllü OG, Giray B, Oztürk M, Favier A et al (2011b) Induction of ROS, p53, p21 in DEHP- and MEHP-exposed LNCaP cells-protection by selenium compounds. Food Chem Toxicol 49:1565–1571

    Article  CAS  Google Scholar 

  • Fan J, Traore K, Li W, Amri H, Huang H, Wu C et al (2010) Molecular mechanisms mediating the effect of mono-(2-ethylhexyl) phthalate on hormone-stimulated steroidogenesis in MA-10 mouse tumor Leydig cells. Endocrinology 151:3348–3362

    Article  CAS  Google Scholar 

  • Fiorini C, Tilloy-Ellul A, Chevalier S, Charuel C, Pointis G (2004) Sertoli cell junctional proteins as early targets for different classes of reproductive toxicants. Reprod Toxicol 18:413–421

    Article  CAS  Google Scholar 

  • Fischer JL, Lancia JK, Mathur A, Smith ML (2006) Selenium protection from DNA damage involves a Ref1/p53/Brca1 protein complex. Anticancer Res 26:899–904

    CAS  Google Scholar 

  • Flohé L (2007) Selenium in mammalian spermiogenesis. Biol Chem 388:987–995

    Article  Google Scholar 

  • Franke WW, Grund C, Schmid E (1979) Intermediate-sized filaments present in Sertoli cells are of the vimentin type. Eur J Cell Biol 19:269–275

    CAS  Google Scholar 

  • Frederiksen H, Skakkebaek NE, Andersson AM (2007) Metabolism of phthalates in humans. Mol Nutr Food Res 51:899–911

    Article  CAS  Google Scholar 

  • Ganther DE (1999) Selenium metabolism, selenoproteins and mechanisms of cancer prevention: complexities with thioredoxin reductase. Carcinogenesis 20:1657–1666

    Article  CAS  Google Scholar 

  • Gazouli M, Yao ZX, Boujrad N, Corton JC, Culty M, Papadopoulos V (2002) Effect of peroxisome proliferators on Leydig cell peripheral-type benzodiazepine receptor gene expression, hormone-stimulated cholesterol transport, and steroidogenesis: role of the peroxisome proliferator-activator receptor alpha. Endocrinology 143:2571–2583

    Article  CAS  Google Scholar 

  • Hall ES, Eveleth J, Boekelheide K (1991) 2, 5-Hexanedione exposure alters the rat Sertoli cell cytoskeleton. II. Intermediate filaments and actin. Toxicol Appl Pharmacol 111:443–453

    Article  CAS  Google Scholar 

  • Hanukoglu I (2006) Antioxidant protective mechanisms against reactive oxygen species (ROS) generated by mitochondrial P450 systems in steroidogenic cells. Drug Metab Rev 38:171–196

    Article  CAS  Google Scholar 

  • Hess RA, Nakai M (2000) Histopathology of the male reproductive system induced by the fungicide benomyl. Histol Histopathol 15:207–224

    CAS  Google Scholar 

  • Kasahara E, Sato EF, Miyoshi M, Konaka R, Hiramoto K, Sasaki J et al (2002) Role of oxidative stress in germ cell apoptosis induced by di(2-ethylhexyl)phthalate. Biochem J 365:849–856

    CAS  Google Scholar 

  • Kaushal N, Bansal MP (2007) Dietary selenium variation-induced oxidative stress modulates CDC2/cyclin B1 expression and apoptosis of germ cells in mice testis. J Nutr Biochem 18:553–564

    Article  CAS  Google Scholar 

  • Kim JM, McGaughy JT, Bogle RK, Ravnik SE (2001) Meiotic expression of the cyclin H/Cdk7 complex in male germ cells of the mouse. Biol Reprod 64:1400–1408

    Article  CAS  Google Scholar 

  • Kluwe WM (1982) Overview of phthalate ester pharmacokinetics in mammalian species. Environ Health Perspect 45:3–9

    Article  CAS  Google Scholar 

  • Koch HM, Preuss R, Angerer J (2006) Di(2-ethylhexyl)phthalate (DEHP): Human metabolism and internal exposure―an update and latest results. Int J Androl 29:155–165

    Article  CAS  Google Scholar 

  • Kwack SJ, Kim KB, Kim HS, Lee BM (2009) Comparative toxicological evaluation of phthalate diesters and metabolites in Sprague-Dawley male rats for risk assessment. J Toxicol Environ Health A 72:1446–1454

    Article  CAS  Google Scholar 

  • Lyche JL, Gutleb AC, Bergman A, Eriksen GS, Murk AJ, Ropstad E et al (2009) Reproductive and developmental toxicity of phthalates. J Toxicol Environ 12:225–249

    CAS  Google Scholar 

  • Noriega NC, Howdeshell KL, Furr J, Lambright CR, Wilson VS, Gray LE Jr (2009) Pubertal administration of DEHP delays puberty, suppresses testosterone production, and inhibits reproductive tract development in male Sprague-Dawley and Long-Evans rats. Toxicol Sci 111:163–178

    Article  CAS  Google Scholar 

  • Oberley TD, Zhong W, Szweda LI, Oberley LW (2000) Localization of antioxidant enzymes and oxidative damage products in normal and malignant prostate epithelium. Prostate 44:144–155

    Article  CAS  Google Scholar 

  • Olson GE, Winfrey VP, Hill KE, Burk RF (2004) Sequential development of flagellar defects in spermatids and epididymal spermatozoa of selenium-deficient rats. Reproduction 127:335–342

    Article  CAS  Google Scholar 

  • Richburg JH, Boekelheide K (1996) Mono-(2-ethylhexyl) phthalate rapidly alters both Sertoli cell vimentin filaments and germ cell apoptosis in young rat testes. Toxicol Appl Pharmacol 137:42–50

    Article  CAS  Google Scholar 

  • Richburg JH, Nanez A, Gao H (1999) Participation of the Fas-signaling system in the initiation of germ cell apoptosis in young rat testes after exposure to mono-(2-ethylhexyl) phthalate. Toxicol Appl Pharmacol 160:271–278

    Article  CAS  Google Scholar 

  • Roveri A, Casasco A, Maiorino M, Dalan P, Calligaro A, Ursini F (1992) Phospholipid hydroperoxide glutathione peroxidase of rat testis. Gonadotropin dependence and immunocytochemical identification. J Biol Chem 267:6142–6146

    CAS  Google Scholar 

  • Seiler A, Schneider M, Förster H, Roth S, Wirth EK, Culmsee C et al (2008) Glutathione peroxidase 4 senses and translates oxidative stress into 12/15-lipoxygenase dependent- and AIF-mediated cell death. Cell Metab 8:237–248

    Article  CAS  Google Scholar 

  • Sen Gupta R, Sen Gupta E, Dhakal BK, Thakur AR, Ahnn J (2004) Vitamin C and vitamin E protect the rat testes from cadmium-induced reactive oxygen species. Mol Cells 17:132–139

    CAS  Google Scholar 

  • Show MD, Anway MD, Folmer JS, Zikrin BR (2003) Reduced intratesticular testosterone concentration alters the polymerization state of the Sertoli cell intermediate filament cytoskeleton by degradation of vimentin. Endocrinology 144:5530–5536

    Article  CAS  Google Scholar 

  • Steinbrenner H, Sies H (2009) Protection against reactive oxygen species by selenoproteins. Biochim Biophys Acta 1790:1478–1485

    Article  CAS  Google Scholar 

  • Tay TW, Andriana BB, Ishii M, Tsunekawa N, Kanai Y, Kurohmaru M (2007) Disappearance of vimentin in Sertoli cells: A mono(2-ethylhexyl)phthalate effect. Int J Toxicol 26:289–295

    Article  CAS  Google Scholar 

  • Virtanen HE, Rajpert-De Meyts E, Main KM, Skakkebaek NE, Toppari J (2005) Testicular dysgenesis syndrome and the development and occurrence of male reproductive disorders. Toxicol Appl Pharmacol 207:501–505

    Article  CAS  Google Scholar 

  • Vogl AW, Pfeiffer DC, Redenbach DM, Grove BD (1993) Sertoli cell cytoskeleton. In: Russell LD, Griswold MD (eds) The Sertoli cell. Cache River Press, Clearwater, FL, pp 39–86

    Google Scholar 

  • Wang ZQ, Watanabe Y, Toki A, Itano T (2002) Altered distribution of Sertoli cell vimentin and increased apoptosis in cryptorchid rats. J Pediatr Surg 37:648–652

    Article  Google Scholar 

  • Zikrin BR, Santulli R, Awoniyi CA, Ewing LL (1989) Maintenance of advanced spermatogenic cells in the adult rat testis: quantitative relationship to testosterone concentration within the testis. Endocrinology 124:3043–3049

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by Hacettepe University Research Fund [Project No: 0701301001].

Author information

Authors and Affiliations

  1. Faculty of Pharmacy, Department of Toxicology, Hacettepe University, 06100, Ankara, Turkey

    Pinar Erkekoglu, Belma Giray & Filiz Hincal

  2. Faculty of Medicine, Department of Histology and Embryology, Hacettepe University, 06100, Ankara, Turkey

    N. Dilara Zeybek & Esin Asan

Authors
  1. Pinar Erkekoglu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. N. Dilara Zeybek
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Belma Giray
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Esin Asan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Filiz Hincal
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Filiz Hincal.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Erkekoglu, P., Zeybek, N.D., Giray, B. et al. The Effects of Di(2-Ethylhexyl)Phthalate Exposure and Selenium Nutrition on Sertoli Cell Vimentin Structure and Germ-Cell Apoptosis in Rat Testis. Arch Environ Contam Toxicol 62, 539–547 (2012). https://doi.org/10.1007/s00244-011-9712-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00244-011-9712-9

Keywords

Search

Navigation