bmp4−/−;smad5−/− zebrafish larvae exhibit fully penetrant, exacerbated ethanol-induced facial malformations. (A-D) Whole-mount images showing the viscerocranium of zebrafish larvae at 5 dpf that had been exposed to ethanol (Ethanol) or not (Untreated). Cartilage is shown in blue, bone in red. Views are ventral, with the anterior to the left. Scale bar: 100 μm. (A) No impact on facial formation was observed in untreated bmp4−/−;smad5+/− larvae (A). Stereotypical smad5 mutant phenotypes were observed in untreated bmp4−/−;smad5−/− larvae (B). Ethanol does not impact viscerocranial morphology in bmp4−/−;smad5+/− larvae (C), but malformations in ethanol-exposed bmp4−/−;smad5−/− larvae (D) are fully penetrant and ethanol sensitive, resulting in several viscerocranial malformations, thereby recapitulating the severe phenotypes observed in dorsomorphin-treated embryos (Lovely et al., 2016).
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