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Fig. 3

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ZDB-FIG-170413-31

Publication

Carroll et al., 2014 - Estrogen defines the dorsal-ventral limit of VEGF regulation to specify the location of the hemogenic endothelial niche

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Fig. 3

E2 Exposure Impairs Formation of the Hemogenic Endothelial Niche

(A) E2 had a biphasic effect on HSPCs: treatment during hemogenic niche specification (12–23 hpf) decreased runx1, whereas exposure during HSC specification and budding (26–36 hpf) increased HSPCs (n ≥ 75/treatment).

(B) Treatment with E2 had no impact on axial vessel formation as assessed by tie2:GFP (n ≥ 20).

(D) Arterial ephrinb2 was robustly decreased by E2 (77/80); red arrowheads indicate artery, and blue arrowheads indicate vein.

(E) Venous flt4 was increased after E2 exposure (40/53).

(F) scl:GFP was reduced by E2 treatment (25/30).

(G) FACS of flk1:GFP+ embryos (n = 32) revealed no change in endothelial cell number by E2 (error bars indicate SD).

(H) qPCR confirmed decreased expression of flk1, ephrinB2, and scl following E2 treatment (mean of triplicate experiments ± SEM; one-tailed t test; flt4^∗p < 0.05, flk1^∗∗p < 0.01, and ephrinB2^∗∗∗p < 0.001).

(I) Injection of scl mRNA increased runx1 expression in the AGM and rescued loss following E2 treatment (n > 55 per treatment).

Expression Data

Genes:	efnb2a EGFP flt4 kdrl myb runx1 tal1
Fish:	WT s849Tg zf255Tg
Condition:	chemical treatment by environment: 17beta-estradiol
Anatomical Terms:	dorsal aorta intersegmental vessel posterior cardinal vein ventral wall of dorsal aorta whole organism
Stage:	Prim-25

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	WT zf255Tg
Condition:	chemical treatment by environment: 17beta-estradiol
Observed In:	dorsal aorta intersegmental vessel posterior cardinal vein ventral wall of dorsal aorta whole organism
Stage:	Prim-25

Phenotype Detail

Acknowledgments

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Reprinted from Developmental Cell, 29, Carroll, K.J., Esain, V., Garnaas, M.K., Cortes, M., Dovey, M.C., Nissim, S., Frechette, G.M., Liu, S.Y., Kwan, W., Cutting, C.C., Harris, J.M., Gorelick, D.A., Halpern, M.E., Lawson, N.D., Goessling, W., North, T.E., Estrogen defines the dorsal-ventral limit of VEGF regulation to specify the location of the hemogenic endothelial niche, 437-53, Copyright (2014) with permission from Elsevier. Full text @ Dev. Cell