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Fig. 7. A transient requirement for MyoD. (A) Embryos were unilaterally co-injected with mRNA coding for nucLacZ together with either MyoD AMO or the equivalent amount of control MO. The injected side is marked with an asterix. (B and E) are injected with control MO, (A, C, D, and F) are injected with MyoD AMO. In situ hybridisation shows Tbx6 (A) and mesogenin (D) expression. (A and D) are gastrula stage 11, viewed vegetally. (B, C, E, and F) are neurula stage 19 and viewed dorsally. (G) qPCR shows levels of target gene expression in sibling embryos at gastrula stage 11(blue bars) and neurula stage 14 (red bars). All qPCR data were normalized to ODC TM and the Ct values from three technical replicates and primer efficiency were then exported into REST confidence interval of the mean fold change. (H) In situ hybridisation shows Delta2 expression in embryos co-injected with mRNA coding for nucLacZ together with either MyoD AMO or control MO. (H, K, and N) are control MO injected, while (I, J, L, M, and O) are injected with MyoD AMO. (H) are open neural plate stage 13, viewed dorsally, and Delta expression shows the first segmentation of somites. (K) are stage 19, viewed dorsally. (N and O) are stage 30 viewed laterally and injected side is shown. Arrows indicate forming somites. (P) A frontal section through a Stage 21 (7 somites) embryo that has been unilaterally co-injected with membrane-RFP and MyoD AMOs (asterix). The structure of the somite cells is revealed by staining with anti-beta-catenin (green) and nuclei are marked with DAPI (blue). The forming somites are numbered on the control side for reference. On the control side, we have highlighted a single cell in the PSM/somitomere, S0, SI and SII to draw attention to the rotation that occurs. On the MyoD knock down side we also highlight cells at progressive points in their rotation; we were unable to highlight a cell in the PSM as the cells were disorganized (arrow heads). Image published in: Maguire RJ et al. (2012) Copyright © 2012. Image reproduced with permission of the Publisher, Elsevier B. V. Gene Synonyms Species Stage(s) Tissue tbx6.L XTbx6 X. laevis Throughout NF stage 11 mesoderm involuting marginal zone tbx6.L XTbx6 X. laevis Throughout NF stage 19 mesoderm presomitic mesoderm msgn1.L mes1, mesogenin, MesP-related bHLH factor, Mespo, msgn1-a, msgn1-b, pMesogenin1 X. laevis Throughout NF stage 11 mesoderm involuted dorsal mesoderm msgn1.L mes1, mesogenin, MesP-related bHLH factor, Mespo, msgn1-a, msgn1-b, pMesogenin1 X. laevis Throughout NF stage 19 mesoderm presomitic mesoderm dlc.L delta-2, delta2, dll3, x-delta-2, X-Delta2 X. laevis Throughout NF stage 13 somite presomitic mesoderm dlc.L delta-2, delta2, dll3, x-delta-2, X-Delta2 X. laevis Throughout NF stage 19 somite presomitic mesoderm dlc.L delta-2, delta2, dll3, x-delta-2, X-Delta2 X. laevis Throughout NF stage 29 and 30 somite presomitic mesoderm Image source: Published Experiment + Assay Source Phenotypes and Disease Xla Wt + myod1 MO + NF11.5 (in situ hybridization) fig.7.a Expression Phenotype decreased amount tbx6.L expression in mesoderm Xla Wt + myod1 MO + NF11.5 (in situ hybridization) fig.7.d Expression Phenotype decreased amount msgn1.L expression in mesoderm Xla Wt + myod1 MO + NF13 (in situ hybridization) fig.7.i, j Expression Phenotype decreased amount dlc.L expression in paraxial mesoderm Xla Wt + myod1 MO + NF19 (in situ hybridization) fig.7.l, m Anatomical Phenotype abnormally anteriorly mislocalised somite Expression Phenotype mislocalised dlc.L expression in somite Xla Wt + myod1 MO + NF19 (in situ hybridization) fig.7.o Anatomical Phenotype abnormally irregular spatial pattern of somite Xla Wt + myod1 MO + NF21 (immunohistochemistry) fig.7.p Anatomical Phenotype abnormal myocyte morphology abnormal presomitic mesoderm morphology abnormally localised myocyte abnormally localised nucleus decreased size of the myocyte decreased size of the presomitic mesoderm Larger Image Printer Friendly View

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