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???displayArticle.abstract??? Neural crest (NC) precursors are stem cells that are capable of forming many cell types after migration to different locations in the embryo. NC and placodes form at the neural plate border (NPB). The Wnt pathway is essential for specifying NC versus placodal identity in this cell population. Here we describe the BTB domain-containing protein Potassium channel tetramerization domain containing 15 (Kctd15) as a factor expressed in the NPB that efficiently inhibits NC induction in zebrafish and frog embryos. Whereas overexpression of Kctd15 inhibited NC formation, knockdown of Kctd15 led to expansion of the NC domain. Likewise, NC induction by Wnt3a plus Chordin in Xenopus animal explants was suppressed by Kctd15, but constitutively active beta-catenin reversed Kctd15-mediated suppression of NC induction. Suppression of NC induction by inhibition of Wnt8.1 was rescued by reduction of Kctd15 expression, linking Kctd15 action to the Wnt pathway. We propose that Kctd15 inhibits NC formation by attenuating the output of the canonical Wnt pathway, thereby restricting expansion of the NC domain beyond its normal range.
Fig. 3. Kctd15 inhibits the output of canonical Wnt signaling. (A) foxd3 expression in control embryos. (B) Loss of foxd3 expression in wnt8.1 morphants. (C) Injection of Wnt8.1 MO together with Kctd15a/15b MOs rescued foxd3 expression. (D) Quantification of A-C; number of embryos (n) is given above bars. (E-J) foxd3 expression in embryos treated with LiCl (E-G). Arrow in G indicates expanded expression in 26/30 embryos. (H-J) LiCl did not rescue foxd3 expression in kctd15a mRNA-injected embryos (I, 40/40; J, 50/50; numbers indicate embryos with appearance as shown in the figure). (K-M) Inhibition of foxd3 by kctd15a (K) was rescued by β-cat* mRNA (M, 41/50 showing foxd3 expression in NC domain). β-cat* mRNA alone induced ectopic expression of foxd3 (L, arrow, 25/31). (A-C,E-M) 1-somite stage; dorsal views, anterior towards top. Scale bar: 100 μm.
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