Figure 6

Chemical inhibition of Na/K ATPase rescues the BPA induced phenotype. (A) 50 hpf control embryo with two normal otolith. (B) Treatment with 1.8 mM ouabain from 6 hpf onwards leads to one misformed otolith. (C) Embryo treated with 70 μM BPA from 5 hpf onwards leads to otolith aggregates. (D) Co treatment with 70 μM BPA and 1.8 mM oubain from 6 hpf onwards leads to normal otolith. (E) Co treatment with 70 μM BPA and 0.9 mM oubain from 6 hpf onwards leads to a mild BPA like phenotype. In some cases one almost normal otolith is found (arrow). (F) Co treatment with 50 μM BPA and 1.8 mM oubain from 6 hpf onwards leads to two normal otoliths. (G) α1a1 morphants display a complete absence of otolith. (H) 70 μM BPA of α1a1 morphants did not rescue otolith formation and result in a complete absence of otolith. (I) α1a1 is expressed in the lower membrane of the otic vesicle at 24 hpf. (J) A similar expression of α1a1 is detected in BPA treated embryos.