Assessment of estrogenic chemicals using an estrogen receptor α (ERα)- and ERβ-mediated reporter gene assay in fish
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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.
KeywordsEstrogen Receptor Genistein Reporter Gene Assay Estrogenic Chemical DDBJ Accession
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).
- Felsenstein J (1995) “PHYLIP (Phlogeny Inference Package),” Version 3.573c. Distributed by author, University of WashingtonGoogle 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 23:167–176CrossRefGoogle 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 373–404Google Scholar