FIGURE

Fig. 3

ID

ZDB-FIG-170227-13

Publication

De Angelis et al., 2017 - Tmem2 Regulates Embryonic Vegf Signaling by Controlling Hyaluronic Acid Turnover

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Fig. 3

Hyaluronic Acid Is Increased in Tmem2 Mutants

(A) Schematic representing the strategy for the HA biosensor. mRNA encoding the HA-binding domain of mouse Neurocan fused to GFP (ssNcan-GFP) was injected into 1- to 2-cell stage embryos, grown, and imaged at the relevant time points.

(B) Lateral views and zoom-in of 30 hpf Tg(kdrl:mCherry) embryos injected with ssNcan-GFP. White arrowheads indicate expression surrounding the axial vessels in the trunk and black arrowhead expression in the heart. Scale bar, 100 μm for whole embryo and 25 μm for the zoom.

(C) ssNcan-GFP is increased and expanded in tmem2^hu5935 mutants compared with siblings around the major axial vessels during sprouting angiogenesis. Maximum intensity projections. Scale bar, 15 μm.

(D) Quantification showing significantly higher fluorescence intensity of ssNcan-GFP in mutant (n = 15) compared with sibling (n = 15) embryos.

(E) Cross-sections of embryos histochemically stained for HA, counterstained for neural red, show increased HA accumulation around the major vessels of 48 hpf tmem2^hu5935 mutant.

(F) Maximum intensity projections of immunofluorescence staining for fibronectin in sibling and tmem2^hu5935 mutants, where blue=low and red=high. Scale bar, 15 μm.

(G) Quantification of fibronectin fluorescence intensity showing no significant difference between mutant (n = 8) and sibling (n = 7) embryos.

DA, dorsal aorta; PCV, posterior cardinal vein; NC, notochord. ^∗∗∗∗p < 0.0001. Error bars represent ±SEM.

Expression Data

Gene:	EGFP
Fish:	s843Tg cemip2^{hu5935/hu5935}; s843Tg
Anatomical Terms:	dorsal aorta endothelial cell posterior cardinal vein endothelial cell
Stage:	Prim-5

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	cemip2^{hu5935/hu5935}
Observed In:	dorsal aorta hyaluronic acid posterior cardinal vein hyaluronic acid
Stage:	Long-pec

Phenotype Detail

Acknowledgments

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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