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ZIRC
ZFIN ID: ZDB-FIG-160608-16
Chen et al., 2016 - Cyp2aa9 regulates haematopoietic stem cell development in zebrafish. Scientific Reports   6:26608 Full text @ Sci. Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Prim-5 to Day 4
PHENOTYPE:
Fish:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Day 4

Fig. 1

cyp2aa9 is required for the HSCs formation.

(a-f) In the cyp2aa9 morphants, the expression of the definitive HSC precursors marker runx1 became decreased in the ventral floor of the dorsal aorta from 25 hpf to 36 hpf. (g,h) The number of cmyb-expressing HSCs was significantly reduced in cyp2aa9 morphants at 36 hpf. (i-l) In contrast to the control embryos at 26 hpf (i, n = 22/24) and 30 hpf (k, n = 23/24), the number of HSCs undergoing budding process from VDA (arrowheads) dramatically reduced in the cyp2aa9 morphant (j, n = 21/25; l, n = 21/25; Scale bar, 50 µm). (m) Quantifications of the number of HSCs undergoing budding process (n = 8, mean ± SD, ***p < 0.001, Student’s t-test). (n,o) In CHT (white brackets), the number of cmyb:GFP-positive cells was significantly reduced in cyp2aa9 morphants (27.6 ± 5.3, mean ± s.e.m., n = 6) comparing with the control (58.2 ± 4.1, mean ± s.e.m., n = 6). Scale bar, 100 µm. (p-s) Fluorecently labelled HSCs and DIC image showed the round-shaped HSCs (white arrowheads) were hardly detected in the CHT (white brackets) of cyp2aa9 morphants under the Tg(CD41:GFP) background (p, n = 24/24; q, n = 25/29; scale bar, 100 µm). CHT regions were zoomed in and showed in the (r,s) (scale bar, 10 µm). (t,u) The expression of T-lymphocyte marker rag1 in the thymus (arrowheads) dramatically decreased in the cyp2aa9 morphants. (a-s) Lateral views, anterior left, dorsal up. (t,u) Ventral views, anterior up.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
EGFP la2Tg standard conditions Protruding-mouth post-vent region IFL
la2Tg + MO1-cyp2aa3,cyp2aa9 standard conditions Protruding-mouth Not Detected post-vent region IFL
zf169Tg standard conditions Long-pec post-vent region IFL
zf169Tg + MO1-cyp2aa3,cyp2aa9 standard conditions Long-pec post-vent region IFL
myb WT standard conditions Prim-25 hematopoietic stem cell ISH
WT + MO1-cyp2aa3,cyp2aa9 standard conditions Prim-25 dorsal aorta hematopoietic stem cell ISH
rag1 WT standard conditions Day 4 thymus ISH
WT + MO1-cyp2aa3,cyp2aa9 standard conditions Day 4 thymus ISH
runx1 WT standard conditions Prim-5 dorsal aorta hematopoietic stem cell ISH
Prim-25 dorsal aorta hematopoietic stem cell ISH
WT + MO1-cyp2aa3,cyp2aa9 standard conditions Prim-5 dorsal aorta hematopoietic stem cell ISH
Prim-25 dorsal aorta hematopoietic stem cell ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO1-cyp2aa3,cyp2aa9 standard conditions Prim-5 dorsal aorta hematopoietic stem cell runx1 expression decreased amount, abnormal
Prim-25 dorsal aorta hematopoietic stem cell runx1 expression decreased amount, abnormal
Prim-25 dorsal aorta hematopoietic stem cell myb expression decreased amount, abnormal
Day 4 thymus rag1 expression decreased amount, abnormal
la2Tg + MO1-cyp2aa3,cyp2aa9 standard conditions Protruding-mouth post-vent region EGFP expression absent, abnormal
Protruding-mouth post-vent region hematopoietic stem cell absent, abnormal
s896Tg; zf169Tg + MO1-cyp2aa3,cyp2aa9 standard conditions Prim-5 dorsal aorta hematopoietic stem cell decreased amount, abnormal
zf169Tg + MO1-cyp2aa3,cyp2aa9 standard conditions Long-pec post-vent region EGFP expression decreased amount, abnormal
Long-pec post-vent region hematopoietic stem cell decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Scientific Reports for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Sci. Rep.