A novel FoxD3 gene trap line reveals neural crest precursor movement and a role for FoxD3 in their specification

Neural crest cells migrate extensively and contribute to diverse derivatives, including the craniofacial skeleton, peripheral neurons and glia, and pigment cells. Although several transgenic lines label neural crest subpopulations, few are suited for studying early events in neural crest development. Here, we present a zebrafish gene/protein trap line gt(foxd3-citrine)ct110a that expresses a Citrine fusion protein with FoxD3, a transcription factor expressed in premigratory and migrating neural crest cells. In this novel line, citrine expression exactly parallels endogenous foxd3 expression. High-resolution time-lapse imaging reveals the dynamic phases of precursor and migratory neural crest cell movements from the neural keel stage to times of active cell migration. In addition, Cre-recombination produces a variant line FoxD3-mCherry-pA whose homozygosis generates a FoxD3 mutant. Taking advantage of the endogenously regulated expression of FoxD3-mCherry fusion protein, we directly assess early effects of FoxD3 loss-of-function on specification and morphogenesis of dorsal root ganglia, craniofacial skeleton and melanophores. These novel lines provide new insights and useful new tools for studying specification, migration and differentiation of neural crest cells.

► A novel zebrafish gene trap line expressing Citrine-tagged endogenous FoxD3.
► Citrine expression recapitulates endogenous FoxD3 expression.
► Cre-recombination produces a FoxD3 mutant.
► Mutant expresses FoxD3-Cherry fusion protein.
► Cherry expression allows analysis of FoxD3 loss-of-function at cellular resolution.