Fig. 8

Human CYP1B1 and zebrafish cyp1b1 genes are evolutionarily and functionally conserved. Live images and methylacrylate sections of 96 hpf Tg(sox10:EGFP) embryos demonstrated that the injection of human CYP1B1 mRNA disrupted neural crest–derived jaw and pharyngeal arch formation and inhibited ocular fissure closure (B, B') compared with controls (A, A'). The knockdown of endogenous Cyp1b1 restored fissure closure (C') and partially rescued the jaw and pharyngeal arch defects resulting from the injection of human CYP1B1 mRNA (C). The injection of human CYP1B1 mRNA containing clinically relevant g.3976G>C (D, D'), g.4490G>A (E, E'), or g.8168G>A (F, F') mutations did not disrupt eye or neural crest development. Western blot analysis (G) showed expression of human CYP1B1 protein (61 kDa) in embryos injected with human CYP1B1 mRNA sequence, but not in uninjected embryos or embryos injected with zebrafish cyp1b1 mRNA. β-actin (42 kDa) was used as a loading control.