Genetic evidence for estrogenicity of bisphenol A in zebrafish gonadal differentiation and its signalling mechanism

Bisphenol A (BPA) can induce endocrine disorders in humans and animals. In this study, we used several zebrafish mutants deficient in estrogen production and signalling, which could be valuable for evaluating estrogenic activities and mechanisms of EDCs. With low endogenous estrogens, the all-male aromatase mutant (cyp19a1a-/-) is expected to be more responsive to estrogenic exposure, and mutants of nuclear estrogen receptors (nERs; esr1-/-, esr2a-/- and esr2b-/-) alone or in combination would allow us to evaluate the action mechanisms of estrogenic EDCs. Exposure to BPA could rescue the all-male phenotype of the cyp19a1a-/- mutant, delayed gonadal development in both sexes, resulting in infertility or subfertility, and caused follicle atresia in females and impairment of spermatogenesis in males. To understand the mechanisms of these effects, we tested BPA in cyp19a1a and nER mutants of different combinations. The feminizing effect of BPA on sexual differentiation was dependent on nERs, in particular esr2a. As for males, nERs were also involved in BPA-induced impairment of spermatogenesis. Taken together, with genome editing technology our study provides the most comprehensive genetic evidence for estrogenic activities of BPA in zebrafish and its action mechanisms. This study also establishes a powerful platform for studying other EDCs with estrogenic activity.