Abstract
Canonical Wnt signaling plays an essential role in proper craniofacial morphogenesis, at least partially due to regulation of various aspects of cranial neural crest development. In an effort to gain insight into the etiology of craniofacial abnormalities resulting from Wnt signaling and/or cranial neural crest dysfunction, we sought to identify Wnt-responsive targets during chick cranial neural crest development. To this end, we leveraged overexpression of a canonical Wnt antagonist, Draxin, in conjunction with RNA-sequencing of cranial neural crest cells that have just activated their epithelial–mesenchymal transition (EMT) program. Through differential expression analysis, gene list functional annotation, hybridization chain reaction (HCR), and quantitative reverse transcription polymerase chain reaction (RT-qPCR), we validated a novel downstream target of canonical Wnt signaling in cranial neural crest – RHOB – and identified possible signaling pathway crosstalk underlying cranial neural crest migration. The results reveal novel putative targets of canonical Wnt signaling during cranial neural crest EMT and highlight important intersections across signaling pathways involved in craniofacial development.
Highlights
The neural crest is a multipotent stem cell population in the vertebrate embryo that undergoes coordinated induction, specification, and epithelial–mesenchymal transition (EMT) events to migrate and differentiate into a wide range of cell types
Consistent with established roles of canonical Wnt signaling and Draxin-mediated inhibition during cranial neural crest epithelial– mesenchymal transition (EMT), we observed enrichment of genes associated with transcriptional regulation, cell adhesion, and lipid synthesis, which we have recently shown is important for cell signaling during cranial neural crest EMT (Piacentino et al, 2020)
Using transcriptome profiling of Draxin-responsive targets, we identified likely gene targets of canonical Wnt signaling during cranial neural crest EMT
Summary
The neural crest is a multipotent stem cell population in the vertebrate embryo that undergoes coordinated induction, specification, and epithelial–mesenchymal transition (EMT) events to migrate and differentiate into a wide range of cell types. Many facets of cranial neural crest development are regulated by Wnt signaling (Wu et al, 2003; Yanfeng et al, 2003; Steventon et al, 2009; Milet and Monsoro-Burq, 2012; Simoes-Costa et al, 2015; Rabadán et al, 2016; Hutchins and Bronner, 2018, 2019; Gandhi et al, 2020).
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