Differential activity of BPA, BPAF and BPC on zebrafish estrogen receptors in vitro and in vivo

Caroline Pinto (1) , Ruixin Hao (1) , Marina Grimaldi (2) , Savini Thrikawala (1) , Abdelhay Boulahtouf (2) , Selim Ait-Aissa (3) , François Brion (3) , Jan-Ake Gustafsson (1, 4) , Patrick Balaguer (2) , Maria Bondesson (5)

1 University of Houston
2 IRCM - U1194 Inserm - UM - Institut de Recherche en Cancérologie de Montpellier
3 INERIS - Institut National de l'Environnement Industriel et des Risques
4 Karolinska Institute
5 Indiana University [Bloomington]

The high volume production compound bisphenol A (BPA) is of environmental concern largely because of its estrogenic activity. Consequently, BPA analogues have been synthesized to be considered as replacement molecules for BPA. These analogues need to be thoroughly evaluated for their estrogenic activity. Here, we combined mechanism zebrafish-based assays to examine estrogenic and anti-estrogenic activities of BPA and two of its analogues, bisphenol AF (BPAF) and bisphenol C (BPC) in vitro and in vivo. In vitro reporter cell lines were used to investigate agonistic and antagonistic effects of the three bisphenols on the three zebrafish estrogen receptors. The transgenic Tg(5 × ERE:GFP) and Cyp19a1b-GFP zebrafish lines were then used to analyze estrogenic and anti-estrogenic responses of the three bisphenols in vivo. BPA, BPAF and BPC were agonists with different potencies for the three zebrafish estrogen receptors in vitro. The potent zfERα-mediated activity of BPA and BPAF in vitro resulted in vivo by activation of GFP expression in zebrafish larvae in the heart (zfERα-dependent) at lower concentrations, and in the liver (zfERβ-dependent) at higher concentrations. BPC induced zfERβ-mediated luciferase expression in vitro, and the zfERβ agonism led to activation of GFP expression in the liver and the brain in vivo. In addition, BPC acted as a full antagonist on zfERα, and completely inhibited estrogen-induced GFP expression in the heart of the zebrafish larvae. To summarize, applying a combination of zebrafish-based in vitro and in vivo methods to evaluate bisphenol analogues for estrogenic activity will facilitate the prioritization of these chemicals for further analysis in higher vertebrates as well as the risk assessment in humans.

Mots clés: ZEBRAFISH; BISPHENOL; ESTROGEN RECEPTOR; TRANSGENIC FISH; REPORTER CELL LINES

Domaines: Ecotoxicologie

Caroline Pinto, Ruixin Hao, Marina Grimaldi, Savini Thrikawala, Abdelhay Boulahtouf, et al.. Differential activity of BPA, BPAF and BPC on zebrafish estrogen receptors in vitro and in vivo. Toxicology and Applied Pharmacology, 2019, 380, pp.art. 114709. ⟨10.1016/j.taap.2019.114709⟩. ⟨ineris-03318109⟩