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Exposure to Estrogenic Chemicals Induces Ectopic Expression of vtg in the Testis of Rainbowfish, Melanotaenia fluviatilis


Vitellogenin (Vtg) is the major egg-yolk precursor protein in oviparous organisms normally synthesised only in mature females. In males and juveniles, the vtg gene, although present, is silent, but its hepatic expression may be activated by xenoestrogens. Surprisingly, its induction and potential consequences in non-hepatic tissues remain unexplored. Here we test the hepatic and testicular response of vtg expression in adult male rainbowfish Melanotaenia fluviatilis exposed to either 1, 3, 5 μg/L 17β-estradiol or 100, 500 μg/L 4-n-nonylphenol for 24–96 h. Significant increase in the expression level of vtg mRNA in the liver and testes of exposed males was observed. The early (24 h), sensitive and reliable detection of the vtg induction using qPCR demonstrates the assay’s robustness to monitor xenobiotic exposure particularly in smaller fish like rainbowfish, an emerging indicator species. Whilst, the ectopic induction of vtg mRNA in testes suggests a more complex Vtg pathway.

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  1. Amdam GV, Norberg K, Hagen A, Omholt SW (2003) Social exploitation of vitellogenin. Proc Natl Acad Sci 100:1799–1802

  2. Arukwe A, Goksøyr A (2003) Eggshell and egg yolk proteins in fish: hepatic proteins for the next generation: oogenetic, population, and evolutionary implications of endocrine disruption. Comp Hepatol 2:4–24

  3. Arukwe A, Knudsen FR, Goksøyr A (1997) Fish zona radiata (eggshell) protein: a sensitive biomarker for environmental estrogens. Environ Health Perspect 105:418–422

  4. Balcombe SR, Sheldon F, Capon SJ, Bond NR, Hadwen WL, Marsh N, Bernays SJ (2011) Climate-change threats to native fish in degraded rivers and floodplains of the Murray–Darling Basin, Australia. Mar Freshwater Res 62:1099–1114

  5. Cooper RL, Kavlock RJ (1997) Endocrine disruptors and reproductive development: a weight-of-evidence overview. J Endocrinol 152:59–166

  6. Denslow N, Chow M, Kroll K, Green L (1999) Vitellogenin as a biomarker of exposure for estrogen or estrogen mimics. Ecotoxicol 8:385–398

  7. Ding JL, Ho B, Valotaire Y, Le Guellec K, Lim EH, Tay SP, Lam TJ (1990) Cloning, characterization and expression of vitellogenin gene of Oreochromis aureus (Teleostei, Cichlidae). Biochem Int 20:843–852

  8. Giudice DG, Prisco M, Agnese M, Verderame M, Limatola E, Andreuccetti P (2011) Expression of vitellogenin in the testis and kidney of the spotted ray Torpedo marmorata exposed to 17β-estradiol. Gen Comp Endocrinol 174:318–325

  9. Guellec KL, Lawless K, Valotaire Y, Kress M, Tenniswood M (1988) Vitellogenin gene expression in male rainbow trout (Salmo gairdneri). Gen Comp Endocrinol 71:359–371

  10. Harries JE, Runnalls T, Hill E, Harris CA, Maddix S, Sumpter JP, Tyler CR (2000) Development of a reproductive performance test for endocrine disrupting chemicals using pair-breeding fathead minnows (Pimephales promelas). Environ Sci Technol 34:3003–3011

  11. Hemmer MJ, Bowman CJ, Hemmer BL, Friedman SD, Marcovich D, Kroll KJ, Denslow ND (2002) Vitellogenin mRNA regulation and plasma clearance in male sheepshead minnows, (Cyprinodon variegatus) after cessation of exposure to 17β-estradiol and p-nonylphenol. Aquat Toxicol 58:99–112

  12. Islinger M, Yuan H, Voelkl A, Braunbeck T (2002) Measurement of vitellogenin gene expression by RT-PCR as a tool to identify endocrine disruption in Japanese medaka (Oryzias latipes). Biomarkers 7:80–93

  13. Jobling S, Sheahan D, Osborne JA, Matthiessen P, Sumpter JP (1996) Inhibition of testicular growth in rainbow trout (Oncorhyncus mykiss) exposed to estrogenic alkylphenolic chemicals. Environ Toxicol Chem 15:194–202

  14. Lee KBH, Lim EH, Lam TJ, Ding JL (1992) Vitellogenin diversity in the perciformes. J Exp Zool 264:100–106

  15. Leusch FDL, Van Den Heuvel MR, Laurie AD, Chapman HF, Gooneratne SR, Trembly LA (2005) Quantification of vitellogenin mRNA induction in mosquitofish (Gambusia affinis) by reverse transcription real-time polymerase chain reaction (RT-PCR). Biomarkers 10:429–438

  16. Muller PY, Janovjak H, Miserez AR, Dobbie Z (2002) Processing of gene expression data generated by quantitative real time PCR. Biotechniques 32:1372–1378

  17. Patil JG, Gunasekera RM (2008) Tissue and sexually dimorphic expression of ovarian and brain aromatase mRNA in the Japanese medaka (Oryzias latipes): implications for their preferential roles in ovarian and neural differentiation and development. Gen Comp Endocrinol 158:131–137

  18. Pollino CA, Georgiades E, Holdway DA (2007) Use of the Australian crimson-spotted rainbowfish (Melanotaenia fluviatilis) as a model test species for investigating the effects of endocrine disruptors. Environ Toxicol Chem 10:2171–2178

  19. Ponza P (2006) Molecular markers of ecotoxicological interest in the rainbowfish, Melanotaenia fluviatilis. PhD thesis. RMIT University

  20. Shanthanagouda AH, Patil JG, Nugegoda D (2012) Ontogenic and sexually dimorphic expression of cyp19 isoforms in the rainbowfish, Melanotaenia fluviatilis (Castelnau 1878). Comp Biochem Physiol A 161:250–258

  21. Shanthanagouda AH, Patil JG, Nugegoda D (2013) Effects of exposure to oestrogenic compounds on aromatase gene expression are gender dependent in the rainbowfish, Melanotaenia fluviatilis. Comp Biochem Physiol C 157:162–171

  22. Shi X, Zhang S, Pang Q (2006) Vitellogenin is a novel player in defence reaction. Fish Shellfish Immunol 20:769–772

  23. Woods M (2007) Assessment of xenoestrogens in the Australian freshwater environment: use, development and validation of in vitro and in vivo models. PhD thesis. University of South Australia

  24. Woods M, Kumar A (2011) Vitellogenin induction by 17b-estradiol and 17a-ethynylestradiol in male Murray rainbowfish (Melanotaenia fluviatilis). Environ Toxicol Chem 30:2620–2627

  25. Yamaguchi A, Ishibashi H, Kohra S, Arizona K, Tominaga N (2005) Short-term effects of endocrine-disrupting chemicals on the expression of estrogen-responsive genes in male medaka (Oryzias latipes). Aquat Toxicol 72:239–249

  26. Yang FX, Xu Y, Hui Y (2006) Reproductive effects of prenatal exposure to nonylphenol on zebrafish (Danio rerio). Comp Biochem Physiol Part C 142:77–84

  27. Yoon SH, Itoh Y, Kaneki G, Nakaniwa M, Ohta M, Watabe S (2008) Molecular characterization of Japanese sillago vitellogenin and changes in its expression levels on exposure to 17beta-estradiol and 4-tert-octylphenol. Mar Biotechnol 10:19–30

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AHS received an RMIT University International Scholarship (RUIS). Authors thank anonymous reviewers for providing useful comments on an earlier draft of the manuscript.

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Correspondence to J. G. Patil.

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Shanthanagouda, A.H., Nugegoda, D., Hassell, K.L. et al. Exposure to Estrogenic Chemicals Induces Ectopic Expression of vtg in the Testis of Rainbowfish, Melanotaenia fluviatilis . Bull Environ Contam Toxicol 91, 438–443 (2013). https://doi.org/10.1007/s00128-013-1078-4

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  • Australian native
  • Xenoestrogen
  • Hepatic
  • Model species