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Figure 2. Cnn2 Is Required for NCC Migration in Frogs and Chicks (A–E) Frog. MO and mRNA were injected unilaterally into left animal blastomeres at the four- to eight-cell stage. Specimens were analyzed for expression of Xtwist by WMISH at the stages indicated. (A–C) NCC migration (A) was compromised upon Cnn2 knockdown (B) and rescued by coinjection of Cnn2 mRNA (C). Histological sections (A′–C′) at the levels indicated (dashed lines) demonstrate the equal (A′ and C′) and unequal (B′) dorsoventral extension of Xtwist-positive NCCs. (D) Qualitative evaluation of results. Embryos were scored as WT or defective. (E) Quantitative evaluation of results. Distances of migration were measured on the injected and uninjected sides as indicated in (B). A box plot of injected/uninjected sides is shown. Note that knockdown and rescue were significant in both cases. (F–L) Chick. Fluorescent MOs (green) were unilaterally electroporated (asterisks) into the neural tube at the four- to eight-somite stage and analyzed for migratory NCCs by HNK1 staining (red) 16 hr thereafter. Migration of cranial (F) and cervical NCCs (G) was unaffected when coMO was injected, but was inhibited on the injected sides upon Ccnn2MO application (I and J). (H and K) Histological sections at cervical levels of specimens treated with coMO (H) or Ccnn2MO (K). (L) Quantitative evaluation of results. HNK1-positive cells were counted on the injected and uninjected sides of coMO- and Ccnn2MO-injected embryos (n = 6 each). Note that Cnn2 knockdown resulted in a reduction of HNK1-positives cells to ∼40%. Error bars represent SD. NT, neural tube; PM, paraxial mesoderm. See also Figure S2. Image published in: Ulmer B et al. (2013) Copyright © 2013. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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