ZFIN ID: ZDB-FIG-190910-26
Bonkhofer et al., 2019 - Blood stem cell-forming haemogenic endothelium in zebrafish derives from arterial endothelium. Nature communications   10:3577 Full text @ Nat. Commun.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-15 to Day 6
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-15 to Day 6

Fig. 6

Aortic HE maintains arterial features in the absence of runx1. a qRT-PCR gene expression analysis of the arterial marker efnb2aand the venous marker ephb4b in the HE and ARE of runx1+/+ (WT) and the HE of runx1−/− mutant (MUT) embryos. Graphs show the mean of detected expression levels relative to the geometric mean (GM) of the two housekeeping genes eef1a1l and rpl13an = 5 independent biological experiments for WT embryos and n = 6 independent biological experiments for MUT embryos. Error bars represent the SEM. Two-way ANOVA; *p < 0.05; **p < 0.01; ***p < 0.001. b Heatmaps of whole-genome gene expression data from RNA-seq for the HE of control and runx1 MO embryos showing arterial and venous genes. cRepresentative fluorescent microscopy image of 6 dpf TgBAC(runx1P2:Citrine), Tg(kdrl:mCherry) double transgenic embryos on a runx1+/+ or runx1−/− genetic background. Left: Region depicting the region of definitive haematopoietic niches including the thymus (Th) and the kidney (Ki). Right: region depicting the DA and the beginning of the caudal haematopoietic tissue (CHT). Green arrows point to the ventral wall of the DA. d Quantification of embryos derived from the indicated in-crosses (InX) depicting the runx1−/− mutant phenotype as depicted in (c). e ISH analysis of runx1 in control and dll4 MO embryos. Experiment was performed with two different MOs. Green arrows point to the HE. f Offspring of dll4+/− heterozygous fish were analysed by ISH for runx1 at 32 hpf. Runx1 expression was quantified by image analysis and subsequent genotyping57. Representative examples of runx1 expression in each of the WT, dll4+/− and dll4−/− genotypes are shown in the right-hand panels and the corresponding expression values are shown as orange points in the graph. (μwt = 33.3, μmut = 15.19, p < 0.0001, F(2,50) = 16,79, one way ANOVA). g Model depicting the lineage relationship between the definitive HE giving rise to HSCs and arterial endothelium in the DA during embryogenesis. Dll4+ aortic progenitors get exposed to localised signals patterning the DA and inducing haemogenic gene expression. Haemogenic specification includes upregulation of runx1 expression and a subsequent Runx1 induced repression of dll4, thus giving rise to Dll4 mature HE

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
Citrine ox1Tg; s896Tg standard conditions Day 6 caudal hematopoietic tissue IFL
Day 6 kidney IFL
Day 6 thymus IFL
Day 6 Not Detected ventral wall of dorsal aorta IFL
runx1hg1/hg1; ox1Tg; s896Tg standard conditions Day 6 caudal hematopoietic tissue IFL
Day 6 Not Detected kidney IFL
Day 6 Not Detected thymus IFL
Day 6 ventral wall of dorsal aorta IFL
runx1 WT standard conditions Prim-15 ventral wall of dorsal aorta ISH
WT + MO1-dll4 standard conditions Prim-15 ventral wall of dorsal aorta ISH
WT + MO4-dll4 standard conditions Prim-15 ventral wall of dorsal aorta ISH
dll4sa9436/sa9436 standard conditions Prim-15 ventral wall of dorsal aorta ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-dll4 standard conditions Prim-15 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
WT + MO4-dll4 standard conditions Prim-15 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
dll4sa9436/sa9436 standard conditions Prim-15 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
runx1hg1/hg1; ox1Tg; s896Tg standard conditions Day 6 caudal hematopoietic tissue Citrine expression decreased amount, abnormal
Day 6 kidney Citrine expression absent, abnormal
Day 6 thymus Citrine expression absent, abnormal
Day 6 ventral wall of dorsal aorta Citrine expression increased amount, abnormal
Acknowledgments:
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