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tfap2axenopus   

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Experiment details for tfap2a

Lee YH et al. (2004) Assay

Early requirement of the transcriptional activator Sox9 for neural crest specification in Xenopus.

Gene Clone Species Stages Anatomy
tfap2a.L laevis NF stage 17 neural crest , cranial neural crest

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  Fig. 4. Sox9 is required for neural crest induction. (A) Schematic representation of the hormone-inducible inhibitory mutant Sox9 construct. Sox9 lacking the transactivation domain (Sox9DC) is fused to the ligand-binding domain of human glucocorticoid receptor (hGR, purple). (B) Experimental timeline. Two-cell stage embryos are injected in one blastomere with 1 ng of Sox9DC-GR mRNA. Embryos are subsequently incubated with dexamethasone (+DEX) at different time point during development (stages 6, 8, 10, 12, or 14), and fixed at stage 17 for detection of Slug, Pax3, or AP2 by whole-mount in situ hybridization. (C) Whole-mount in situ hybridization of embryos injected in one blastomere at the two-cell stage with 1 ng of Sox9DC-GR mRNA and treated with dexamethasone (+DEX) at the gastrula (stage 10) or neurula (stage 14) stages. Embryos were analyzed for expression of three early neural crest markers (Slug, Pax3, and AP2) at stage 15/17. Activation of Sox9 inhibitory mutant (Sox9DC-GR) before stage 14 blocks formation of neural crest progenitors. RNA encoding the lineage tracer h-galactosidase was coinjected to identify the injected side (red staining, left side in all panels, arrows). Embryos are viewed from the dorsal side, anterior to the top. (D) Quantification of Slug, Pax3, and AP2 in situ hybridization results. The numbers at the top of each bar indicate the number of cases analyzed. Slug (red bars) and Pax3 (blue bars) expressions were analyzed after treatment with dexamethasone at five time points, stages 6, 8, 10, 12, and 14. AP2 expression (green bars) was analyzed after treatment with dexamethasone at three time points, stages 6, 10, and 14.

Gene Clone Species Stages Anatomy
tfap2a.L laevis NF stage 25 pharyngeal arch , mandibular arch , hyoid arch , branchial arch , cranial neural crest

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  Fig. 5. Neural crest migration occurs independently of Sox9 function. (A) Experimental timeline. Two-cell stage embryos are injected in one blastomere with 1 ng of Sox9DC-GR mRNA. Embryos are subsequently incubated with dexamethasone (+Dex) at different time points during migration of the neural crest (stages 17, 20, or 22), and fixed at stage 25 for detection of Twist and AP2 in the migrating cranial neural crest by whole-mount in situ hybridization. (B) AP2 and Twist whole-mount in situ hybridization of such embryos. Activation of Sox9 inhibitory mutant (+Dex) after neural crest specification (stage 17 or 22) does not prevent cranial neural crest cell migration. Note that the extent of migration of AP2-and Twist-expressing cells in the cranial region is identical in control (uninjected) and injected sides. RNA encoding the lineage tracer h-galactosidase was coinjected to identify the injected side (red staining). Embryos are viewed from the lateral side, anterior to the right (left panels) or anterior to the left (right panels).