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ZFIN ID: ZDB-FIG-170227-11
De Angelis et al., 2017 - Tmem2 Regulates Embryonic Vegf Signaling by Controlling Hyaluronic Acid Turnover. Developmental Cell   40:123-136 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibody:
Fish:
Anatomical Terms:
Stage Range: 26+ somites to Long-pec
PHENOTYPE:
Fish:
Observed In:
Stage Range: 26+ somites to Long-pec

Fig. 1

Tmem2 Is Required for Angiogenesis and Vegf Signaling

(A) Lateral composite views of sibling and tmem2hu5935 mutant embryos crossed with the Tg(kdrl:mCherry) line (white) show a failure to complete angiogenesis at both arterial (28 hpf) and venous (50 hpf) sprouting stages. Yellow and white asterisks indicate the absence of sprouts in the mutants. Scale bar, 100 μm for whole embryo and 25 μm for zoom.

(B) Column scatterplots depicting the number of segments with T-branching arterial ISVs per embryo at 28 hpf (n = 17 siblings, n = 29 mutants).

(C) Scatterplots depicting the number of segments with venous ISVs per embryo at 50 hpf (n = 10 siblings and n = 11 mutants).

(D and E) Maximum projection for whole-mount pErk immunofluorescence staining (white and heatmap) in sibling and tmem2hu5935 mutants on the Tg(kdrl:EGFP) background (green) at (D) 22 hpf and (E) 32 hpf. Average projections of fluorescence intensity for pErk are represented by heatmaps, where blue=low and red=high. Scale bar, 10 μm. pErk-positive cells indicated with white arrowheads; open arrowheads depict staining in corpuscle of Stannius.

(F and G) Quantification of pErk-positive endothelial cells across four somites showed (F) significantly fewer pErk-positive endothelial cells in tmem2hu5935 mutant embryos compared with siblings at 22 hpf during primary sprouting (n = 8 sibling, n = 8 mutants) and (G) significantly lower fluorescence intensity (normalized to neuronal expression) of pERK-positive endothelial cells in mutants (n = 37) compared with siblings (n = 57) at 22 hpf.

(H and I) Similarly, (H) there were significantly fewer pERK-positive endothelial cells in the PCV across eight somites of mutants (n = 8) at 32 hpf compared with siblings (n = 8; p < 0.01) and (I) significantly lower fluorescence intensity of pERK-positive cells in mutants (n = 20) compared with siblings (n = 52) at 32 hpf.

DA, dorsal aorta; PCV, posterior cardinal vein. ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Error bars represent ±SD.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP cemip2hu5935/hu5935; s843Tg standard conditions 26+ somites dorsal aorta endothelial cell IHC
Prim-15 dorsal aorta endothelial cell IHC
Prim-15 posterior cardinal vein endothelial cell IHC
s843Tg standard conditions 26+ somites dorsal aorta endothelial cell IHC
Prim-15 dorsal aorta endothelial cell IHC
Prim-15 posterior cardinal vein endothelial cell IHC
mCherry cemip2hu5935/hu5935; s916Tg standard conditions Prim-5 dorsal aorta IFL
Prim-5 intersegmental vessel IFL
Prim-5 posterior cardinal vein IFL
Prim-5 vasculature IFL
Long-pec dorsal aorta IFL
Long-pec intersegmental artery IFL
Long-pec intersegmental vein IFL
Long-pec intersegmental vessel IFL
Long-pec posterior cardinal vein IFL
Long-pec vasculature IFL
s916Tg standard conditions Prim-5 dorsal aorta IFL
Prim-5 intersegmental vessel IFL
Prim-5 posterior cardinal vein IFL
Prim-5 vasculature IFL
Long-pec dorsal aorta IFL
Long-pec intersegmental artery IFL
Long-pec intersegmental vein IFL
Long-pec intersegmental vessel IFL
Long-pec posterior cardinal vein IFL
Long-pec vasculature IFL
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab9-mapk IHC cemip2hu5935/hu5935; s843Tg standard conditions 26+ somites dorsal aorta endothelial cell
IHC Prim-15 posterior cardinal vein endothelial cell
IHC s843Tg standard conditions 26+ somites dorsal aorta endothelial cell
IHC Prim-15 posterior cardinal vein endothelial cell
Phenotype Details
Fish Conditions Stage Phenotype
cemip2hu5935/hu5935; s843Tg standard conditions 26+ somites dorsal aorta endothelial cell ab9-mapk labeling decreased amount, abnormal
26+ somites dorsal aorta endothelial cell ab9-mapk labeling decreased distribution, abnormal
Prim-15 posterior cardinal vein endothelial cell ab9-mapk labeling decreased amount, abnormal
Prim-15 posterior cardinal vein endothelial cell ab9-mapk labeling decreased distribution, abnormal
cemip2hu5935/hu5935; s916Tg standard conditions Prim-5 intersegmental artery decreased amount, abnormal
Prim-5 intersegmental vessel sprouting angiogenesis decreased occurrence, abnormal
Long-pec intersegmental vein decreased amount, abnormal
Long-pec intersegmental vessel sprouting angiogenesis decreased occurrence, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Cell for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

Reprinted from Developmental Cell, 40, De Angelis, J.E., Lagendijk, A.K., Chen, H., Tromp, A., Bower, N.I., Tunny, K.A., Brooks, A.J., Bakkers, J., Francois, M., Yap, A.S., Simons, C., Wicking, C., Hogan, B.M., Smith, K.A., Tmem2 Regulates Embryonic Vegf Signaling by Controlling Hyaluronic Acid Turnover, 123-136, Copyright (2017) with permission from Elsevier. Full text @ Dev. Cell