ZFIN ID: ZDB-FIG-170413-32
Carroll et al., 2014 - Estrogen defines the dorsal-ventral limit of VEGF regulation to specify the location of the hemogenic endothelial niche. Developmental Cell   29:437-53 Full text @ Dev. Cell
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Conditions:
Anatomical Terms:
Stage Range: 14-19 somites to Prim-25
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage Range: 14-19 somites to Prim-25

Fig. 4

Exogenous E2 Signaling Disrupts VEGF/NOTCH Cascades in the AGM

(A) E2 exposure had no impact on deltaC expression by WISH at 18 hpf but strongly reduced arterial expression at 24 and 36 hpf (n > 30/treatment); an arrowhead indicates artery.

(B) Expression of notch3 was similarly regulated by E2 at 18, 24, and 36 hpf (n > 30/treatment); an arrowhead indicates artery.

(C) qPCR confirmed E2-mediated reductions in deltaC and notch3 expression at 36 hpf (mean of triplicate experiments ± SEM; one-tailed t test deltaC, notch3p < 0.05).

(D) Decreased Notch:GFP was observed in the artery (white bracket indicates artery walls) after E2 treatment from 12–36 or 12–23 hpf; no change occurred with exposure from 26–36 hpf (n ≥ 52).

(E) NICD induction increased expression of runx1/cmyb and rescued HSPCs in E2-treated embryos (n ≥ 31/treatment).

(F) Expression of VEGFAa was strongly decreased in zebrafish following E2 treatment at both 18 and 24 hpf (n ≥ 25/treatment); an arrow points to somite staining.

(G) qPCR confirmed that expression of VEGFAa was decreased at 24 hpf (mean of triplicate experiments ± SEM; one-tailed t test ∗∗∗p < 0.001).

(H) Induction of VEGFAa increased runx1 expression and rescued the effect of E2 exposure; treatment with DAPT decreased expression of runx1 and blocked the VEGF-mediated rescue of HSPCs (n ≥ 20/treatment); an arrowhead indicates artery.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
dlc WT control 14-19 somites whole organism ISH
Prim-5 dorsal aorta ISH
Prim-25 dorsal aorta ISH
WT chemical treatment by environment: 17beta-estradiol 14-19 somites whole organism ISH
Prim-5 dorsal aorta ISH
Prim-25 dorsal aorta ISH
Prim-25 whole organism RTPCR
EGFP um14Tg control Prim-25 dorsal aorta IFL
um14Tg chemical treatment by environment: 17beta-estradiol Prim-25 dorsal aorta IFL
myb kca3Tg; kca4Tg control Prim-25 ventral wall of dorsal aorta ISH
kca3Tg; kca4Tg chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta ISH
kca3Tg; kca4Tg heat shock Prim-25 ventral wall of dorsal aorta ISH
kca3Tg; kca4Tg heat shock, chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta ISH
notch3 WT control 14-19 somites whole organism ISH
Prim-5 dorsal aorta ISH
Prim-25 dorsal aorta ISH
WT chemical treatment by environment: 17beta-estradiol 14-19 somites whole organism ISH
Prim-5 dorsal aorta ISH
Prim-25 dorsal aorta ISH
Prim-25 whole organism RTPCR
runx1 kca3Tg; kca4Tg control Prim-25 ventral wall of dorsal aorta ISH
kca3Tg; kca4Tg chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta ISH
kca3Tg; kca4Tg heat shock Prim-25 ventral wall of dorsal aorta ISH
kca3Tg; kca4Tg heat shock, chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta ISH
nc2aTg control Prim-25 ventral wall of dorsal aorta ISH
nc2aTg chemical treatment by environment: DAPT Prim-25 ventral wall of dorsal aorta ISH
nc2aTg chemical treatment by environment: DAPT, chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta ISH
nc2aTg chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta ISH
nc2aTg heat shock, chemical treatment by environment: DAPT, chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta ISH
nc2aTg heat shock, chemical treatment by environment: DAPT Prim-25 ventral wall of dorsal aorta ISH
nc2aTg heat shock Prim-25 ventral wall of dorsal aorta ISH
nc2aTg heat shock, chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta ISH
vegfaa WT control 14-19 somites somite ISH
Prim-5 somite ISH
WT chemical treatment by environment: 17beta-estradiol 14-19 somites somite ISH
Prim-5 somite ISH
Prim-5 whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT chemical treatment by environment: 17beta-estradiol 14-19 somites somite vegfaa expression decreased amount, abnormal
Prim-5 dorsal aorta dlc expression decreased amount, abnormal
Prim-5 dorsal aorta notch3 expression decreased amount, abnormal
Prim-5 somite vegfaa expression decreased amount, abnormal
Prim-5 whole organism vegfaa expression decreased amount, abnormal
Prim-25 dorsal aorta dlc expression decreased amount, abnormal
Prim-25 dorsal aorta notch3 expression decreased amount, abnormal
Prim-25 whole organism dlc expression decreased amount, abnormal
Prim-25 whole organism notch3 expression decreased amount, abnormal
kca3Tg; kca4Tg chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta myb expression decreased amount, abnormal
Prim-25 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
kca3Tg; kca4Tg heat shock Prim-25 ventral wall of dorsal aorta myb expression increased amount, abnormal
Prim-25 ventral wall of dorsal aorta runx1 expression increased amount, abnormal
kca3Tg; kca4Tg heat shock, chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta myb expression amount, ameliorated
Prim-25 ventral wall of dorsal aorta runx1 expression amount, ameliorated
nc2aTg chemical treatment by environment: DAPT Prim-25 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
nc2aTg chemical treatment by environment: DAPT, chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
nc2aTg chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
nc2aTg heat shock, chemical treatment by environment: DAPT, chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
nc2aTg heat shock, chemical treatment by environment: DAPT Prim-25 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
nc2aTg heat shock Prim-25 ventral wall of dorsal aorta runx1 expression increased amount, abnormal
nc2aTg heat shock, chemical treatment by environment: 17beta-estradiol Prim-25 ventral wall of dorsal aorta runx1 expression amount, ameliorated
um14Tg chemical treatment by environment: 17beta-estradiol Prim-25 dorsal aorta EGFP expression decreased amount, 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, 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