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nkx2-1xenopus   

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Experiment details for nkx2-1

Rankin SA et al. (2014) Assay

A Molecular atlas of Xenopus respiratory system development.

Gene Clone Species Stages Anatomy
nkx2-1.L laevis NF stage 33 and 34 brain , forebrain , endoderm , eye , foregut , [+]
nkx2-1.L laevis NF stage 42 endoderm , trachea , foregut , lung primordium
nkx2-1.L laevis NF stage 49 endoderm , trachea , foregut , lung primordium
nkx2-1.L laevis NF stage 59 endoderm , trachea , lung , foregut , lung primordium

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  Figure 3. Molecular characterization of the developing Xenopus foregut and respiratory epithelium. A: Schematic of the developing foregut region and respiratory system at the indicated developmental stages, with endoderm in yellow, heart (h) in orange, and respiratory epithelium in blue. B–Y: Whole-mount in situ hybridization with the indicated probes to whole embryos at NF32 (B,F,J), NF33 (N), NF35 (R), NF38 (V) or to isolated guts at NF42 (C,G,K,O,S,W) or to lung/tracheal/esophageal regions dissected at NF49 (D,H,L,P,T,X) or NF59 (E,I,M,Q,U,Y). Abbreviations: e, esophagus; fg, foregut; h, heart; lb, lung bud; lg, lung; lv, liver; st, stomach; tr, trachea; ty, thyroid. Scale bar = 0.5 mmDownload figure to PowerPoint

Gene Clone Species Stages Anatomy
nkx2-1.L laevis NF stage 33 and 34 endoderm , foregut , ventral , lung primordium
nkx2-1.L laevis NF stage 35 and 36 endoderm , foregut , ventral , lung primordium
nkx2-1.L laevis NF stage 42 trachea , foregut , lung primordium , epithelial cell

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  Figure 4. Immunofluorescence analysis of D–V foregut patterning and trachea-esophagus separation in Xenopus. A–O: Confocal immunostaining of the Xenopus foregut showing transverse sections at three positions along the anterior–posterior axis at the indicated stages showing protein expression of Sox2 (blue), Nkx2.1 (red), and FoxF1 (green). P: Transverse section through the E9.5 mouse foregut showing similar expression of Sox2 (blue) Nkx2.1 (red) and FoxF1 (green). Q,R: Transverse sections of the Xenopus foregut at NF36 and NF42 showing P63 expression (white) in the dorsal foregut (Q) and esophagus (R). Abbreviations: dfg, dorsal foregut; eso, esophagus; fge, foregut epithelium; h, heart; lb, lung bud; lpm, lateral plate mesoderm; nc, notochord; s, somite; st, stomach; tr, trachea; ubb, ultimobrachial body; vfg, ventral foregut. Scale bar = 0.1 mm.Download figure to PowerPoint

Gene Clone Species Stages Anatomy
nkx2-1.L laevis NF stage 33 and 34 to NF stage 35 and 36 forebrain , telencephalon , diencephalon , endoderm , eye , [+]

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  Fig. 10. Functional epistasis of Wnt/b-catenin, FGF, and BMP pathways. A–O: Wnt/b-catenin is necessary and sufficient downstream of FGF to induce respiratory fate. Embryos were injected at the 4-cell stage in both dorsal anterior blastomeres with control-MOs or wnt2/2b-MOs (10 ng/ blastomere) and then injected at the 8-cell stage dorsal-vegetally with 800 pg iFGFR1 mRNA (400 pg/blastomere). Subsequently embryos were treated from stages NF25–NF35 with either, DMSO, PD173074 (100 mM), DMH1 (20 mM), or AP20187 (2 mM to induce the iFGFR1 construct activ- ity). Embryos were analyzed at NF34/35 by in situ hybridization for nkx2.1 expression (A, D, G, J, and M) or by for Nkx2.1 (red), Sox2 (blue), and FoxF1 (green) (B, E, H, K, N). At NF42 siblings were analyzed by in situ hybridization for sftpc expression (C, F, I, L, O). P–Y: The expansion of respiratory fate by Wnt/b-catenin requires BMP signaling. Embryos were injected at the 4-cell stage in both dorsal-anterior blastomeres with control-MOs or wnt2/2b-MOs (10 ng/blastomere) and then injected at the 8-cell stage dorsal-vegetally with 800 pg GR-LefdN-BCTA mRNA (400 pg/blastomere). Subsequently the embryos were treated with DMSO, DMH-1, or PD173047 from NF25-35, and then with dex from NF28-35 and analyzed by in situ hybridization for nkx2.1 at NF35 (P,R,T,V,X) and sftpc at NF42 (Q,S,U,W,Y).