ZFIN ID: ZDB-FIG-161111-4
Cortes et al., 2016 - Developmental Vitamin D Availability Impacts Hematopoietic Stem Cell Production. Cell Reports   17:458-468 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: Prim-25
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Prim-25

Fig. 1

VDR Is Necessary for Zebrafish Definitive Hematopoiesis

(A) Representative runx1 WISH images and qualitative phenotype distribution (high/medium/low, n ≥ 30 embryos/condition) of vdra MO-injected embryos and sibling controls at 36 hpf. Error bars, mean ± SD.

(B) WISH example and phenotype distribution for flk1 in vdra morphants (n ≥ 20 embryos/condition).

(C) qPCR analysis quantifying the impact on HSPCs (runx1, ∗∗p < 0.01) and hemogenic endothelium (flk1, ephrinb2a, p = N.S.) in control and vdra morphants (30 embryos/sample × 3 replicates).

(D-F) Representative WISH (D), phenotype distributions (E), and qPCR quantification (F) of vdra mutant embryos (runx1, ∗∗p < 0.01; flk1, ephrinb2a, p = N.S.) phenocopied the impact on HSPCs but not on vessels.

(G) FACS analysis of vdra morphants indicated significant reductions in Flk1:dsRed+/cMyb:GFP+ HSPCs (p < 0.05) at 48 hpf (5 embryos/sample × 4 replicates/condition). Error bars, mean ± SD.

(H) WISH samples and phenotype distribution of runx1/cmyb expression in cyp27b1 MO-injected embryos at 36 hpf (n ≥ 30 embryos/condition).

(I) FACS analysis showed a significant decrease in HSPCs (∗∗p < 0.01) with cyp27b1 MO injection (n value and error bars as in C).

Scale bars, 100 μm. See also Figure S1.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
kdrl WT standard conditions Prim-25 ventral wall of dorsal aorta ISH
Prim-25 whole organism RTPCR
WT + MO2-vdra standard conditions Prim-25 ventral wall of dorsal aorta ISH
Prim-25 whole organism RTPCR
vdrazf735/zf735 standard conditions Prim-25 ventral wall of dorsal aorta ISH
Prim-25 whole organism RTPCR
runx1 WT standard conditions Prim-25 ventral wall of dorsal aorta ISH
Prim-25 whole organism RTPCR
WT + MO2-vdra standard conditions Prim-25 ventral wall of dorsal aorta ISH
Prim-25 whole organism RTPCR
vdrazf735/zf735 standard conditions Prim-25 ventral wall of dorsal aorta ISH
Prim-25 whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-cyp27b1 standard conditions Prim-25 hematopoietic stem cell proliferation disrupted, abnormal
Prim-25 ventral wall of dorsal aorta has fewer parts of type hematopoietic multipotent progenitor cell, abnormal
WT + MO2-vdra standard conditions Prim-25 hematopoietic stem cell proliferation disrupted, abnormal
Prim-25 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
Prim-25 ventral wall of dorsal aorta has fewer parts of type hematopoietic multipotent progenitor cell, abnormal
Prim-25 whole organism runx1 expression decreased amount, abnormal
pd27Tg; sd2Tg; zf169Tg; zf71Tg standard conditions Prim-25 ventral wall of dorsal aorta has normal numbers of parts of type hematopoietic multipotent progenitor cell, normal
pd27Tg; sd2Tg; zf169Tg; zf71Tg + MO1-cyp27b1 standard conditions Prim-25 hematopoietic stem cell proliferation disrupted, abnormal
Prim-25 ventral wall of dorsal aorta has fewer parts of type hematopoietic multipotent progenitor cell, abnormal
pd27Tg; sd2Tg; zf169Tg; zf71Tg + MO2-vdra standard conditions Prim-25 hematopoietic stem cell proliferation disrupted, abnormal
Prim-25 ventral wall of dorsal aorta has fewer parts of type hematopoietic multipotent progenitor cell, abnormal
vdrazf735/zf735 standard conditions Prim-25 hematopoietic stem cell proliferation disrupted, abnormal
Prim-25 ventral wall of dorsal aorta runx1 expression decreased amount, abnormal
Prim-25 ventral wall of dorsal aorta has fewer parts of type hematopoietic multipotent progenitor cell, abnormal
Prim-25 whole organism runx1 expression decreased amount, abnormal
Acknowledgments:
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