Xenbase Image ID: 80905

Figure 3. Co(III)-Ebox inhibits Snail-mediated neural crest cell specification in Xenopus via a temperature-dependent mechanism. (A) In situ hybridization of Sox10 expression showing neural crest formation (left) and Twist expression showing neural crest migration (right) of embryos injected with Co(III)-Ebox into one cells of embryos at the 8-cell stage. Embryos were co-injected with β-galactosidase mRNA as a lineage tracer (red staining). Neural crest forming/migrating regions indicated with arrowheads on the injected side. (B, C) In situ hybridization showing the neural crest of control embryos or embryos injected with Co(III)-sb, Ebox, Co(III)-EboxMut or Co(III)-Ebox and grown at 14 (B) or 27 (C). Top panel: whole mount in situ hybridization showing FoxD3 expression in the neural crest and Krox20 expression in rhombomeres 3/5 of the CNS. For FoxD3 embryos reared at 14, Co(III)-sb n = 12; Ebox n = 16; Co(III)-EboxMut n = 16; and Co(III)-Ebox n = 13. For FoxD3 embryos reared at 27, Co(III)-sb = 13; Ebox n = 18; Co(III)-EboxMut n = 14; and Co(III)-Ebox n = 18. Bottom panel: expression of Sox10 in the neural crest and Krox20 (blue arrowheads). For Sox10 embryos reared at 14, Co(III)-sb n = 21; Ebox n = 33; Co(III)-EboxMut n = 35; and Co(III)-Ebox n = 38. For Sox10 embryos reared at 27, Co(III)-sb = 33; Ebox n = 35; Co(III)-EboxMut n = 35; and Co(III)-Ebox n = 36. Loss of Sox10 and FoxD3 is seen in the neural crest forming regions of embryos treated with Co(III)-Ebox grown at 27 (red arrowheads). (D) Cartoon representation of expected in situ hybridization result for Krox20 and neural crest markers (Sox10 and FoxD3) at stage 17. Black staining represents neural crest forming regions visualized by in situ hybridization for Sox10 or FoxD3 and indicated by red arrows. Pale blue staining indicates hindbrain forming regions of rhombomeres 3 and 5 visualized by in situ hybridization with Krox20 and indicated by blue arrows. (E) Cartoon representation of expected in situ hybridization result for Krox20 and neural crest markers (Sox10 and Twist) during neural crest cell migration. Black staining represents migrating neural crest regions visualized by in situ hybridization for Sox10 or Twist and indicated by red arrows. Pale blue staining indicates hindbrain forming regions of rhombomeres 3 and 5 visualized by in situ hybridization with Krox20 and indicated by blue arrows. (F) Co(III)-Ebox incubated in embryo lysate for 0 to 24 hours retains the and ability to inhibit XSnai2 DNA binding as visualized by EMSA. Image published in: Harney AS et al. (2012) Harney et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Gene Synonyms Species Stage(s) Tissue sox10.L X. laevis Throughout NF stage 17 neural crest cranial neural crest chordal neural plate border sox10.L X. laevis Throughout NF stage 24 neural crest migratory neural crest cell cranial neural crest mandibular crest hyoid crest anterior branchial crest posterior branchial crest egr2.L EGR-2, Krox-20, krox20, XKr20, XKrox-20 X. laevis Throughout NF stage 17 rhombomere R3 rhombomere R5 foxd3.L fkh6, foxd3-a, foxd3-b, XFD-6, xfoxd3 X. laevis Throughout NF stage 17 neural crest cranial neural crest twist1.S twist, twist1-a, twist1-b, X-twi, Xtwi, Xtwist X. laevis Throughout NF stage 19 neural crest cranial neural crest migratory neural crest cell Image source: Published Larger Image Printer Friendly View

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