|
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
|
Display additional annotations [+]
Gene |
Clone |
Species |
Stages |
Anatomy |
foxa2
|
|
laevis
|
NF stage 32
|
spinal cord
,
endoderm
,
head endoderm
,
foregut
,
head
|
foxa2
|
|
laevis
|
NF stage 42
|
endoderm
,
trachea
,
esophagus
,
liver
,
foregut
,
[+]
|
foxa2
|
|
laevis
|
NF stage 49
|
endoderm
,
trachea
,
lung
,
esophagus
,
stomach
,
[+]
|
foxa2
|
|
laevis
|
NF stage 59
|
endoderm
,
respiratory system
,
trachea
,
lung
,
foregut
,
[+]
|
sox2
|
|
laevis
|
NF stage 32
|
brain
,
endoderm
,
eye
,
foregut
,
head
,
[+]
|
sox2
|
|
laevis
|
NF stage 42
|
endoderm
,
trachea
,
foregut
,
lung primordium
|
sox2
|
|
laevis
|
NF stage 49
|
endoderm
,
trachea
,
esophagus
,
foregut
,
lung primordium
|
sox2
|
|
laevis
|
NF stage 59
|
lung primordium
|
shh
|
|
laevis
|
NF stage 32
|
brain
,
notochord
,
endoderm
,
pharyngeal arch
,
liver diverticulum
,
[+]
|
shh
|
|
laevis
|
NF stage 42
|
endoderm
,
foregut
,
lung primordium
,
liver primordium
|
shh
|
|
laevis
|
NF stage 49
|
endoderm
,
trachea
,
esophagus
,
stomach
,
foregut
,
[+]
|
shh
|
|
laevis
|
NF stage 59
|
trachea
,
lung
,
esophagus
|
sftpc
|
|
laevis
|
NF stage 37 and 38
|
endoderm
,
foregut
,
lung primordium
,
epithelial cell
|
sftpc
|
|
laevis
|
NF stage 42
|
endoderm
,
trachea
,
lung
,
foregut
,
epithelial cell
|
sftpc
|
|
laevis
|
NF stage 49
|
endoderm
,
trachea
,
lung
,
foregut
,
epithelial cell
|
sftpc
|
|
laevis
|
NF stage 59
|
endoderm
,
trachea
,
lung
,
foregut
,
epithelial cell
|
sox9
|
|
laevis
|
NF stage 35 and 36
|
brain
,
notochord
,
eye
,
head
,
pharyngeal region
|
sox9
|
|
laevis
|
NF stage 42
|
endoderm
,
foregut
,
lung primordium
|
sox9
|
|
laevis
|
NF stage 49
|
endoderm
,
trachea
,
lung
,
esophagus
,
foregut
|
sox9
|
|
laevis
|
NF stage 59
|
endoderm
,
trachea
,
lung
,
esophagus
,
foregut
|
|
|
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 |
|
Display additional annotations [+]
Gene |
Clone |
Species |
Stages |
Anatomy |
foxf1
|
|
laevis
|
NF stage 32
|
lateral plate mesoderm
|
foxf1
|
|
laevis
|
NF stage 33 and 34
|
lateral plate mesoderm
|
foxf1
|
|
laevis
|
NF stage 35 and 36
|
mesoderm
|
foxf1
|
|
laevis
|
NF stage 42
|
mesoderm
,
trachea
,
esophagus
,
stomach
,
smooth muscle
|
sox2
|
|
laevis
|
NF stage 32
|
endoderm
,
foregut
,
dorsal
,
epithelium
,
epithelial cell
|
sox2
|
|
laevis
|
NF stage 33 and 34
|
endoderm
,
foregut
,
dorsal
|
sox2
|
|
laevis
|
NF stage 35 and 36
|
endoderm
,
esophagus
,
stomach
,
foregut
,
pharynx
,
[+]
|
sox2
|
|
laevis
|
NF stage 42
|
esophagus
,
stomach
,
foregut
,
epithelial cell
|
|
|
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 |
|
Display additional annotations [+]
|
|
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). |