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Figure 5. Temperature modulation of Co(III)-Ebox-mediated Snail inhibition reveals a requirement for Snail in melanocyte formation. Neurula stage embryos injected with Co(III)-Ebox or Co(III)-EboxMut in the archenteron space were maintained at 14 and (A) neural crest migration visualized by whole mount in situ hybridization for Twist expression. Krox20 is used to visualize rhombomeres 3/5 in the hindbrain. Untreated n = 8; Co(III)-EboxMut n = 8; and Co(III)-Ebox n = 12. (B) Quantification of mandibular, hyoid and branchial streams of neural crest cells are represented as the distance migrated as a percentage of control embryos. T-tests determined that neither Co(III)-EboxMut nor Co(III)-Ebox had a significant effect on neural crest cell migration. (C) Sibling embryos were maintained until swimming tadpole stages at 14 (top panel) or 27 (bottom panel) when melanocyte formation could be assessed. Arrowheads point to regions of diminished melanocytes. For embryos maintained at 14, untreated n = 12; Co(III)-EboxMut n = 12; and Co(III)-Ebox n = 12. For embryos maintained at 27, untreated n = 12; Co(III)-EboxMut n = 12; and Co(III)-Ebox n = 13. (D) Same embryos were maintained until swimming tadpole stages at 14 (top panel) or 27 (bottom panel) when craniofacial cartilage formation could be assessed. Cartliage was stained with Alcian blue and dissected to visualize cartilage formation. (E) Injected embryos were reared at 14 (top panel) or 27 (bottom panel) until stage 28 and glial cell formation was visualized by whole mount in situ hybridization for FoxD3 expression. For embryos maintained at 14, untreated n = 15; Co(III)-EboxMut n = 17; and Co(III)-Ebox n = 11. For embryos maintained at 27, untreated n = 15; Co(III)-EboxMut n = 15; and Co(III)-Ebox n = 22. (F) Injected embryos were reared at 14 (top panel) or 27 (bottom panel) until stage 28 and primary neuron formation was visualized by whole mount in situ hybridization for N-tubulin expression. For embryos maintained at 14, untreated n = 15; Co(III)-EboxMut n = 17; and Co(III)-Ebox n = 13. For embryos maintained at 27, untreated n = 20; Co(III)-EboxMut n = 21; and Co(III)-Ebox n = 21. 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 twist1.S twist, twist1-a, twist1-b, X-twi, Xtwi, Xtwist X. laevis Throughout NF stage 24 cranial neural crest mandibular crest hyoid crest branchial crest egr2.L EGR-2, Krox-20, krox20, XKr20, XKrox-20 X. laevis Throughout NF stage 24 hindbrain rhombomere R3 rhombomere R5 foxd3.L fkh6, foxd3-a, foxd3-b, XFD-6, xfoxd3 X. laevis Throughout NF stage 28 cranial nerve central nervous system spinal cord nerve tubb2b.S N-Tub, n-tubulin, NBT, neural beta-tubulin, NST, ntubulin, Xn-tubulin X. laevis Throughout NF stage 28 neuron cranial nerve nerve cephalic nerve central nervous system brain spinal cord Image source: Published Larger Image Printer Friendly View

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