FIGURE

Fig. 3

ID
ZDB-FIG-141217-3
Publication
Choe et al., 2014 - Tbx1 controls the morphogenesis of pharyngeal pouch epithelia through mesodermal Wnt11r and Fgf8a
Other Figures
All Figure Page
Back to All Figure Page
Fig. 3

Rescue of tbx1-/-pouch defects with mesodermal Fgf8a and Wnt11r. (A-J) Fluorescent in situ hybridization for fgf8a or wnt11r (green) and GFP immunohistochemistry to detect nkx2.5:GFP-positive mesoderm (red) at 30hpf. In wild type, fgf8a was expressed in ventral nkx2.5:GFP-positive mesodermal cores of arches 3 and 4 (m3 and m4), whereas wnt11r was in more dorsal subsets of these mesodermal cores. In tbx1 mutants, mesodermal expression of fgf8a (n=12 of 12) and wnt11r (n=10 of 11) was lost, although fgf8a and wnt11r expression in the anterior (arrow) and posterior (arrowheads) portions of the otic vesicle was unaffected. An nkx2.3:Tbx1 transgene did not restore mesodermal fgf8a (n=0 of 7) and wnt11r expression (n=0 of 5), whereas an nkx2.5:Tbx1 transgene restored mesodermal fgf8a (n=5 of 6) and wnt11r expression (n=9 of 9). Also, nkx2.5:Fgf8-GFP and nkx2.5:Wnt11r transgenes restored fgf8a (n=27 of 27) and wnt11r (n=21 of 21) expression, respectively. (K-O) Alcama immunohistochemistry (green) showed five pouches (p1-p5) in wild-type fish at 34hpf. tbx1-/- mutants lost all pouches except for the first (p1), whereas individual nkx2.5:Wnt11r or nkx2.5:Fgf8a-GFP transgenes modestly rescued, and combined nkx2.5:Wnt11r and nkx2.5:Fgf8a-GFP transgenes strongly rescued posterior pouches (p2-p5). Sensory ganglia are indicated with red asterisks. (P-T) Ventral views of dissected facial cartilages. A bilateral set of five CBs formed in wild-type zebrafish, and no CBs formed in tbx1 mutants. No rescue of CB cartilage was seen in nkx2.5:Wnt11r; tbx1-/- (n=39), nkx2.5:Fgf8a-GFP; tbx1-/- (n=34), or nkx2.5:Wnt11r; nkx2.5:Fgf8a-GFP; tbx1-/- larvae (n=31). (U) Quantification of pouch defects based on Alcama staining in wild type (n=51), nkx2.5:Wnt11r (n=49), nkx2.5:Fgf8a-GFP (n=36), tbx1-/- (n=62), nkx2.5:Wnt11r; tbx1-/- (n=44), nkx2.5:Fgf8a-GFP; tbx1-/- (n=37), and nkx2.5:Wnt11r; nkx2.5:Fgf8a-GFP; tbx1-/- (n=24). Data represent meanĀ±s.e.m., P values are shown. (V) Low magnification view of an embryo at 32hpf showing nkx2.5:GFP-positive mesoderm (green) relative to her5:mCherryCAAX-positive pouch endoderm (red) and the developing eye and ear. The schematic shows expression of fgf8a (green) and wnt11r (yellow) within distinct subsets of nkx2.5-positive mesoderm (red) during the formation of endodermal pouches (blue). Scale bars: 40μm (A-O).

Expression Data
Genes:
Antibody:
Fish:
Anatomical Terms:
Stage: Prim-15

Expression Detail
Antibody Labeling
Phenotype Data
Fish:
Observed In:
Stage: Prim-15

Phenotype Detail
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Development