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Assessment of estrogenic chemicals using an estrogen receptor α (ERα)- and ERβ-mediated reporter gene assay in fish

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Abstract

In vitro reporter gene assays using vertebrate cell lines or yeast have been used for assessment of the estrogenic chemicals. However, estrogen receptor α (ERα)- and ERβ-mediated reporter gene system in fish has yet to be developed. In the present study, we developed an ERα- and ERβ-mediated reporter gene assay in fish and estimated estrogenic activities of 17β-estradiol (E2), p-nonylphenol (NP), bisphenol A (BPA), p,p′-DDE, and genistein (Gen) using the in vitro reporter assay. The activity was intensely induced by transfection with either ERα or ERβ expression plasmid under E2 or Gen administration, whereas it was significantly induced by transfection with ERα expression plasmid, but not with ERβ expression plasmid under NP administration. On the other hand, the activity was induced more intensely by transfection with ERα expression plasmid than ERβ expression plasmid under BPA or p,p′-DDE administration. These results indicate that there are obvious differences in the activity through ERα and ERβ among the estrogenic chemicals examined in vitro. Thus, the in vitro reporter assay provides an excellent system for elucidating the action mechanism of estrogenic chemicals in fishes.

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References

  • Ackermann GE, Brombacher E, Fent K (2002) Development of a fish reporter gene system for the assessment of estrogenic compounds and sewage treatment plant effluents. Environ Toxicol Chem 21:1864–1875

    Article  CAS  Google Scholar 

  • Chang XT, Kobayashi T, Todo T, Ikeuchi T, Yoshiura M, Kajiura-Kobayashi H, Morrey C, Nagahama Y (1999) Molecular cloning of estrogen receptors α and β in the ovary of a teleost fish, the tilapia (Oreochromis niloticus). Zool Sci 16:653–658

    Article  CAS  Google Scholar 

  • Felsenstein J (1995) “PHYLIP (Phlogeny Inference Package),” Version 3.573c. Distributed by author, University of Washington

  • Hawkins MB, Thornton JW, Crew D, Skipper JK, Dotte A, Thomas P (2000) Identification of a third distinct estrogen receptor and reclassification estrogen receptors in teleost. Proc Natl Acad Sci USA 97:10751–10756

    Article  CAS  Google Scholar 

  • Kitano T, Takamune K, Kobayashi T, Nagahama Y, Abe S-I (1999) Suppression of P450 aromatase gene expression in sex-reversed males produced by rearing genetically female larvae at a high water temperature during a period of sex differentiation in the Japanese flounder (Paralichthys olivaceus). J Mol Endocrinol 23:167–176

    Article  CAS  Google Scholar 

  • Kuiper GGJM, Enmark E, Pelto-Huikko M, Nilsson S, Gustafsson JA (1996) Cloning of a novel estrogen receptor expressed in rat prostate and ovary. Proc Natl Acad Sci USA 93:5925–5930

    Article  CAS  Google Scholar 

  • Kuiper GGJM, Lemmen JG, Carlsson B, Corton JC, Safe SH, van der Saag PT, van der Burg B, Gustafsson JA (1998) Interaction of estrogenic chemicals and phytoestrogens with estrogen receptor β. Endocrinology 139:4252–4263

    Article  CAS  Google Scholar 

  • Marin MG, Matozzo V (2004) Vitellogenin induction as a biomarker of exposure to estrogenic compounds in aquatic environments. Mar Pollut Bull 48:835–839

    Article  CAS  Google Scholar 

  • Meerts IATM, Letcher RJ, Hoving S, Marsh G, Bergman Å, Lemmen JG, Van der Burg B, Brouwer A (2001) In vitro estrogenicity of polybrominated diphenyl ethers, hydroxylated PBDEs, and polybrominated bisphenol A compounds. Environ Health Perspect 109:399–407

    Article  CAS  Google Scholar 

  • Menuet A, Pellegrini E, Anglade I, Blaise O, Laudet V, Kah O, Pakdel F (2002) Molecular characterization of three estrogen receptor forms in zebrafish: binding characteristics, transactivation properties, and tissue distributions. Biol Reprod 66:1881–1892

    Article  CAS  Google Scholar 

  • Mosselman S, Polman J, Dijkema R (1996) ERβ: identification and characterization of a novel human estrogen receptor. FEBS Lett 392:49–53

    Article  CAS  Google Scholar 

  • Ng TB, Idler DR (1983) Yolk formation and differentiation in teleost fishes. In: Hoar WS, Randall DJ, Donaldson EM (eds) Fish physiology, vol 9A. Academic, New York, pp 373–404

    Google Scholar 

  • Routledge EJ, Sumpter JP (1996) Estrogenic activity of surfactants and some of their degradation products assessed using a recombinant yeast screen. Environ Toxicol Chem 15:241–248

    Article  CAS  Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  PubMed  Google Scholar 

  • Seiler-Tuyns A, Mérillat AM, Haefliger DN, Wahli W (1988) The human estrogen receptor can regulate exogenous but not endogenous vitellogenin gene promoters in a Xenopus cell line. Nucleic Acids Res 16:8291–8305

    Article  CAS  Google Scholar 

  • Shimasaki Y, Kitano T, Oshima Y, Inoue S, Imada N, Honjo T (2002) Tributyltin causes masculinization in fish. Environ Toxicol Chem 22:141–144

    Article  Google Scholar 

  • Socorro S, Power DM, Olsson P-E, Canario AVM (2000) Two estrogen receptors expressed in the teleost fish, Sparus aurata: cDNA cloning, characterization and tissue distribution. J Endocrinol 166:293–306

    Article  CAS  Google Scholar 

  • Tabata K (1991) Induction of gynogenetic diploid males and presumption of sex determination mechanisms in the hirame, Paralichthys olivaceus. Bull Jpn Soc Sci Fish 57:845–850

    Article  Google Scholar 

  • Teo BY, Tan NS, Lim EH, Lam TJ, Ding JL (1998) A novel piscine vitellogenin gene: structural and functional analyses of estrogen-inducible promoter. Mol Cell Endocrinol 146:103–120

    Article  CAS  Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

    Article  CAS  Google Scholar 

  • Walker P, Germond JE, Brown-Luedi M, Givel F, Wahli W (1984) Sequence homologies in the region preceding the transcription initiation site of the liver estrogen-responsive vitellogenin and apo-VLDLII genes. Nucleic Acids Res 12:8611–8626

    Article  CAS  Google Scholar 

  • White R, Jobling S, Hoare SA, Sumpter JP, Parker MG (1994) Environmentally persistent alkylphenolic compounds are estrogenic. Endocrinology 135:175–182

    Article  CAS  Google Scholar 

  • Xia Z, Patiño R, Gale WL, Maule AG, Densmore LD (1999) Cloning, in vitro expression, and novel phylogenetic classification of a channel catfish estrogen receptor. Gen Comp Endocrinol 113:360–368

    Article  CAS  Google Scholar 

  • Xia Z, Gale WL, Chang X, Langenau D, Patiño R, Maule AG, Densmore LD (2000) Phylogenetic sequence analysis, recombinant expression, and tissue distribution of a channel catfish estrogen receptor β. Gen Comp Endocrinol 118:139–149

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Professor Yoshitaka Nagahama for helpful advice. This work was supported by CREST (Core Research for Evolutional Science and Technology) of JST (Japan Science and Technology Corporation).

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Correspondence to T. Kitano.

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Communicated by R. Cattaneo-Vietti, Genova

Physical and Chemical Impacts on Marine Organisms, a Bilateral Seminar Italy–Japan held in November 2004

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Kitano, T., Koyanagi, T., Adachi, R. et al. Assessment of estrogenic chemicals using an estrogen receptor α (ERα)- and ERβ-mediated reporter gene assay in fish. Marine Biology 149, 49–55 (2006). https://doi.org/10.1007/s00227-005-0206-z

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