Xenbase Image ID: 81439

Fig. 4. Combined overexpression of Otx2 and Su(H)Ank efficiently induces ectopic Dmrt5 and Dmrt4 expression in the ectoderm. (A)C) Anterior views of neurula stage embryos hybridized with Otx2, Delta1 or Dmrt5. Arrows indicate the region of the olfactory placodes. (D)-L) Otx2-Notch combined activation in the ectoderm efficiently induces Dmrt5 and Dmrt4 expression in the ectoderm. Embryos injected with MT-Otx2-hGR and MT-hGR-Su(H)Ank (500 pg each) were induced with Dex at stage 123, and then subjected to LacZ staining and in situ hybridization with the indicated probes. (D)I) Dorsal views with anterior to the right. (J)L) Lateral views of the injected side. Moderate ectopic Dmrt4 and Dmrt5 expression was detected in embryos injected with Otx2 mRNA alone throughout the ectoderm. While moderate ectopic Dmrt5 expression mainly restricted to the head ectoderm was also detected in Su(H)Ank mRNA injected embryos, no ectopic Dmrt4 was observed in Su(H)Ank mRNA injected embryos. Embryos coexpressing Otx2 and Su(H)Ank show robust ectopic Dmrt4 and Dmrt5 staining throughout the ectoderm. FoxE3 used as control was as previously reported not induced in embryos injected with Otx2 alone or with Su(H)Ank alone injected embryos but was ectopically activated in embryos injected with both constructs. (M) Real time RT-PCR analysis of Dmrt5 and Dmrt4 expression in animal caps derived from four-cell stage embryos injected with MT-Otx2-hGR or MT-hGR-Su(H)Ank mRNA, alone or in combination (500 pg per blastomere). Note that their combination induces stronger expression of Dmrt5 and Dmrt4 than that induced by Otx2 or Su(H)Ank alone. Each value has been normalized to the level of GAPDH expression. (N) Real time RT-PCR analysis of the expression of Dmrt5, Dmrt4 and XAG in animal caps derived from embryos injected with MT-Otx2-hGR and MT-hGR-Su(H)Ank mRNA, treated or not with dexamethasone (Dex) and cycloheximide (CHX). Note that both Dmrt5 and Dmrt4, but not XAG, are activated by MT-Otx2-hGR and MT-hGR-Su(H)Ank in the presence of CHX. Image published in: Parlier D et al. (2013) Copyright © 2013. Image reproduced with permission of the Publisher, Elsevier B. V. Gene Synonyms Species Stage(s) Tissue otx2.S otx-2, otx2-a, otx2-b, otxA, Xotx-2, Xotx2 X. laevis Throughout NF stage 13 neural plate dmrta2.S dmrt5 X. laevis Throughout NF stage 13 neural plate pre-chordal neural plate dll1.L delta-1, Delta-1, delta1, x-delta, X-delta-1, Xdelta-1, Xdelta1, XDelta1 X. laevis Throughout NF stage 13 anterior neural ridge anterior neural fold Image source: Published Larger Image Printer Friendly View

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