ZFIN ID: ZDB-FIG-161130-1
Grainger et al., 2016 - Wnt9a Is Required for the Aortic Amplification of Nascent Hematopoietic Stem Cells. Cell Reports   17:1595-1606 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Conditions:
Anatomical Term:
Stage Range: Prim-15 to Prim-25
PHENOTYPE:
Fish:
Conditions:
Observed In:
Stage Range: Prim-15 to Prim-25

Fig. 1

Wnt Signaling Is Required Transiently Prior to 20 hpf for HSPC Development

(A) kdrl:mCherry; cmyb:eGFP fish were treated with IWP-L6 or LiCl to inhibit and activate Wnt signaling, respectively ( van de Water et al., 2001 and Wang et al., 2013), from 10 hpf to 36 hpf and imaged at 36 hpf. A, aorta; V, vein. Scale bar, 30 μm.

(B) Quantitation of HSPCs per millimeter of aorta.

(C) Schematic of heat shock regimen.

(D) hsp:dntcf fish were heat shocked every hour from 13 hpf to 24 hpf, fixed at 40 hpf, and analyzed for cmyb expression by WISH. Scale bar, 100 μm.

(E) Quantitation of cmyb+ cells from (D).

(F) Schematic of experimental layout.

(G) hsp:dntcf fish were heat shocked at 16.5 hpf, pools were fixed every hour from 23 to 36 hpf, and they were analyzed for cmyb expression by WISH. Scale bar, 100 μm.

(H) Quantitation of cmyb+ cells from (G).

(I) HSPCs are specified as endothelial cells ingress toward the midline. Endothelial cells receive specification cues, at least in part, from somatic cells. At 26 hpf, HSPCs start to emerge from the floor of the dorsal aorta. A, aorta; V, vein.

∗∗p < 0.01; ∗∗∗p < 0.001; n.s., not significant. Error bars represent SD. See also Figure S1.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP y171Tg; zf169Tg control Prim-25 ventral wall of dorsal aorta hematopoietic stem cell IFL
y171Tg; zf169Tg chemical treatment by environment: Wnt signalling inhibitor Prim-25 ventral wall of dorsal aorta hematopoietic stem cell IFL
y171Tg; zf169Tg chemical treatment by environment: lithium chloride Prim-25 ventral wall of dorsal aorta hematopoietic stem cell IFL
mCherry y171Tg; zf169Tg control Prim-25 ventral wall of dorsal aorta hematopoietic stem cell IFL
y171Tg; zf169Tg chemical treatment by environment: Wnt signalling inhibitor Prim-25 ventral wall of dorsal aorta hematopoietic stem cell IFL
y171Tg; zf169Tg chemical treatment by environment: lithium chloride Prim-25 ventral wall of dorsal aorta hematopoietic stem cell IFL
myb w26Tg control Prim-15 ventral wall of dorsal aorta hematopoietic stem cell ISH
Prim-25 ventral wall of dorsal aorta hematopoietic stem cell ISH
w26Tg heat shock Prim-15 ventral wall of dorsal aorta hematopoietic stem cell ISH
Prim-25 ventral wall of dorsal aorta hematopoietic stem cell ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
w26Tg heat shock Prim-15 ventral wall of dorsal aorta hematopoietic stem cell myb expression decreased amount, abnormal
Prim-15 ventral wall of dorsal aorta hematopoietic stem cell decreased amount, abnormal
Prim-25 ventral wall of dorsal aorta hematopoietic stem cell myb expression decreased amount, abnormal
Prim-25 ventral wall of dorsal aorta hematopoietic stem cell decreased amount, abnormal
y171Tg; zf169Tg chemical treatment by environment: Wnt signalling inhibitor Prim-25 ventral wall of dorsal aorta hematopoietic stem cell decreased amount, abnormal
y171Tg; zf169Tg chemical treatment by environment: lithium chloride Prim-25 ventral wall of dorsal aorta hematopoietic stem cell increased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Cell Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Cell Rep.