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ZFIN ID: ZDB-FIG-170829-2
Abrial et al., 2017 - TGF-β Signaling Is Necessary and Sufficient for Pharyngeal Arch Artery Angioblast Formation. Cell Reports   20:973-983 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Terms:
Stage: Prim-25
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage: Prim-25

Fig. 3

TGF-β Signaling Is Required for the PAA Progenitor to Angioblast Transition

(A–D) Left lateral views of etsrp+ PAA angioblasts in 36-hpf embryos treated from 20-hpf stage with DMSO or the indicated, increasing concentrations of LY-364947. The angioblast clusters that did form in LY-364947-treated embryos were smaller and/or failed to sprout (asterisks in C and D). Scale bars, 200 μm.

(E) Bar graph showing the average number of etsrp+ angioblast clusters ±SD within each experimental group (∗∗p < 0.01; ∗∗∗p < 0.001). n = 40–50 embryos per treatment group with a total of four biological replicates.

(F and G) Left lateral views of nkx2.5+ PAA progenitors in 36-hpf embryos treated from 20 hpf with DMSO (F, n = 43) or LY-364947 (G, n = 38/54 with the phenotype shown, 14/54 showed a milder phenotype, 2/54 appeared normal). The line in (G) highlights persistent expression of nkx2.5 transcripts in PAA progenitors of arches 3–6 in LY-364947-treated embryos.

(H and I) Left lateral views of 38-hpf Tg(fli1a:nGFP),Tg(nkx2.5:ZsYellow) embryos treated from 20 hpf with DMSO or LY-364947, immunostained for GFP and ZsYellow, and imaged in the green and red channels to visualize PAA endothelial (pseudocolored magenta) and progenitor (pseudocolored yellow) cell nuclei, respectively.

(J) The bar graph shows the average numbers ±SD of nkx2.5:ZsYellow+ PAA progenitors in DMSO and LY-364947-treated embryos. n = 4 for both experimental groups. ∗∗∗p < 0.0001. Scale bars, 25 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
etsrp WT control Prim-25 pharyngeal arch 3 angioblastic mesenchymal cell ISH
Prim-25 pharyngeal arch 4 angioblastic mesenchymal cell ISH
Prim-25 pharyngeal arch 5 angioblastic mesenchymal cell ISH
Prim-25 pharyngeal arch 6 angioblastic mesenchymal cell ISH
Prim-25 pharyngeal vasculature angioblastic mesenchymal cell ISH
WT chemical treatment by environment: LY 364947 Prim-25 pharyngeal arch 3 angioblastic mesenchymal cell ISH
Prim-25 pharyngeal arch 4 angioblastic mesenchymal cell ISH
Prim-25 pharyngeal arch 5 angioblastic mesenchymal cell ISH
Prim-25 Not Detected pharyngeal arch 6 angioblastic mesenchymal cell ISH
GFP fb7Tg/fb7Tg ; y7Tg control Prim-25 pharyngeal arch 3 blood vessel endothelial cell IHC
Prim-25 pharyngeal arch 4 blood vessel endothelial cell IHC
Prim-25 pharyngeal arch 5 blood vessel endothelial cell IHC
fb7Tg/fb7Tg ; y7Tg chemical treatment by environment: LY 364947 Prim-25 pharyngeal arch 3 blood vessel endothelial cell IHC
Prim-25 pharyngeal arch 4 blood vessel endothelial cell IHC
Prim-25 pharyngeal arch 5 blood vessel endothelial cell IHC
nkx2.3 WT control Prim-25 pharyngeal arch ISH
WT chemical treatment by environment: LY 364947 Prim-25 pharyngeal arch ISH
ZsYellow fb7Tg/fb7Tg ; y7Tg control Prim-25 pharyngeal arch 3 embryonic blood vessel endothelial progenitor cell IHC
Prim-25 pharyngeal arch 4 embryonic blood vessel endothelial progenitor cell IHC
Prim-25 pharyngeal arch 5 embryonic blood vessel endothelial progenitor cell IHC
fb7Tg/fb7Tg ; y7Tg chemical treatment by environment: LY 364947 Prim-25 pharyngeal arch 3 embryonic blood vessel endothelial progenitor cell IHC
Prim-25 pharyngeal arch 4 embryonic blood vessel endothelial progenitor cell IHC
Prim-25 pharyngeal arch 5 embryonic blood vessel endothelial progenitor cell IHC
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT chemical treatment by environment: LY 364947 Prim-25 pharyngeal arch 3 angioblastic mesenchymal cell etsrp expression decreased amount, abnormal
Prim-25 pharyngeal arch 3 angioblastic mesenchymal cell decreased amount, abnormal
Prim-25 pharyngeal arch 4 angioblastic mesenchymal cell decreased amount, abnormal
Prim-25 pharyngeal arch 4 angioblastic mesenchymal cell etsrp expression decreased amount, abnormal
Prim-25 pharyngeal arch 5 angioblastic mesenchymal cell decreased amount, abnormal
Prim-25 pharyngeal arch 5 angioblastic mesenchymal cell etsrp expression decreased amount, abnormal
Prim-25 pharyngeal arch 6 angioblastic mesenchymal cell etsrp expression absent, abnormal
fb7Tg/fb7Tg ; y7Tg chemical treatment by environment: LY 364947 Prim-25 pharyngeal arch 3 blood vessel endothelial cell decreased amount, abnormal
Prim-25 pharyngeal arch 3 blood vessel endothelial cell GFP expression decreased amount, abnormal
Prim-25 pharyngeal arch 3 embryonic blood vessel endothelial progenitor cell ZsYellow expression increased amount, abnormal
Prim-25 pharyngeal arch 3 embryonic blood vessel endothelial progenitor cell increased amount, abnormal
Prim-25 pharyngeal arch 4 blood vessel endothelial cell GFP expression decreased amount, abnormal
Prim-25 pharyngeal arch 4 blood vessel endothelial cell decreased amount, abnormal
Prim-25 pharyngeal arch 4 embryonic blood vessel endothelial progenitor cell increased amount, abnormal
Prim-25 pharyngeal arch 4 embryonic blood vessel endothelial progenitor cell ZsYellow expression increased amount, abnormal
Prim-25 pharyngeal arch 5 blood vessel endothelial cell GFP expression decreased amount, abnormal
Prim-25 pharyngeal arch 5 blood vessel endothelial cell decreased amount, abnormal
Prim-25 pharyngeal arch 5 embryonic blood vessel endothelial progenitor cell ZsYellow expression increased amount, abnormal
Prim-25 pharyngeal arch 5 embryonic blood vessel endothelial progenitor cell increased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Rep.