Restoration of normal Hox code and branchial arch morphogenesis after extensive deletion of hindbrain neural crest

Among the derivatives of the cephalic neural crest is the ectomesenchyme which subsequently constitutes most of the craniofacial skeleton. There is evidence to suggest that the skeletogenic fate of the hindbrain neural crest is specified before emigration from the neural tube and that Antennapedia class Hox genes are involved in that process. To explore the putative causal link between Hox expression and craniofacial morphology, we produced a specific series of bilateral crest deletions in chick embryos and assessed branchial arch morphology, Hox gene expression, and patterning of skeletal structures in the postoperative embryo. Surprisingly, we found that deletion of the bulk of the rhombencephalic crest and substantial portions of the dorsal rhombencephalon did not prevent normal branchial arch morphogenesis and normal patterns of Hox gene (-A3 and -B4) expression 48 h after operation. Neural crest-like cells have been identified on crest migration pathways at the level of the original ablation, further confirming that ablated cephalic neural crest is replaced by regeneration from the cut edge of the neuroepithelium. Furthermore, in such embryos ectomesenchyme from regenerated crest is able to form a facial skeleton in which the mandible and hyoid apparatus are normal in size and organization. These findings demonstrate that the cranial neuroepithelium has more extensive regenerative capacities than was previously thought, which has important implications for investigations of craniofacial development.