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ZIRC
ZFIN ID: ZDB-FIG-160304-60
Hofsteen et al., 2016 - Quantitative proteomics identify DAB2 as a cardiac developmental regulator that inhibits WNT/β-catenin signaling. Proceedings of the National Academy of Sciences of the United States of America   113(4):1002-7 Full text @ Proc. Natl. Acad. Sci. USA
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagent:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Condition:
Knockdown Reagent:
Observed In:
Stage Range: Prim-5 to Long-pec

Fig. 4

Dab2 promotes cardiomyocyte development by negatively regulating WNT/β-catenin signaling. (A) qRT-PCR of dab2 and a panel of known WNT/β-catenin target genes (axin2, tcf7I1a, and tcf3b) and ligands (wnt3, wnt8a, wnt5a, and wnt10a) in control (black bars) and Dab2 mutants (orange bars) at 24 hpf. (B–E) Representative brightest point projection confocal image (Z series) of control (B and C) and Dab2 mutants (D and E) at 24 hpf. TCFSiam:nlsmCherry is shown in red, flk1:GFP in green, and DAPI in blue. Panels shown are anterior merged (B and D), anterior red (B′ and D′), and posterior red and green (C and E). (F) Quantification of total fluorescence intensity between control and Dab2 mutants at 24 hpf. n ≥3 biological replicates. *P ≤ 0.05, Student’s t test. Data are mean ± SEM. (G) Representative brightest point projection confocal (Z series) images of control (Top) and Dab2 mutants (Bottom) at 48 hpf. cmlc2:DsRed-nuc is in red (Left), and ALCAM is in magenta (Middle) (H) Quantification of atrial (At) and ventricular (V) cardiomyocytes. (I) Percent gross morphological defect scoring in control and Dab2 mutants after overexpression of DKK1 scored at 48 hpf. (J) Representative images of normal, mild, and severe heart defects at 48 hpf. (K) Percent heart defects in control and Dab2 mutants after overexpression of DKK1 scored at 48 hpf. (L) Quantification of total cardiomyocyte number. *P ≤ 0.05, Student’s t test. n = 30–45 per group for heat shock and scoring for panels I and K. For quantification of cardiomyocyte nuclei, n = 4–6 fish hearts per condition analyzed by z-series confocal microscopy. Data are mean ± SEM. (M) Model: DAB2 inhibition of WNT/β-catenin signaling promotes cardiomyocyte differentiation.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
axin2 AB control Prim-5 whole organism RTPCR
AB + CRISPR1-dab2 standard conditions Prim-5 whole organism RTPCR
dab2 AB control Prim-5 whole organism RTPCR
AB + CRISPR1-dab2 standard conditions Prim-5 whole organism RTPCR
mCherry ia5Tg/+ ; zn1Tg/+ control Prim-5 forebrain IFL
Prim-5 hindbrain IFL
Prim-5 midbrain IFL
Prim-5 pronephros anterior region IFL
Prim-5 vasculature posterior region IFL
ia5Tg/+ ; zn1Tg/+ + CRISPR1-dab2 standard conditions Prim-5 forebrain IFL
Prim-5 hindbrain IFL
Prim-5 midbrain IFL
Prim-5 pronephros anterior region IFL
Prim-5 vasculature posterior region IFL
tcf3b AB control Prim-5 whole organism RTPCR
AB + CRISPR1-dab2 standard conditions Prim-5 whole organism RTPCR
tcf7l1a AB control Prim-5 whole organism RTPCR
AB + CRISPR1-dab2 standard conditions Prim-5 whole organism RTPCR
wnt3 AB control Prim-5 whole organism RTPCR
AB + CRISPR1-dab2 standard conditions Prim-5 whole organism RTPCR
wnt5a AB control Prim-5 whole organism RTPCR
AB + CRISPR1-dab2 standard conditions Prim-5 whole organism RTPCR
wnt8a AB control Prim-5 whole organism RTPCR
AB + CRISPR1-dab2 standard conditions Prim-5 whole organism RTPCR
wnt10a AB control Prim-5 whole organism RTPCR
AB + CRISPR1-dab2 standard conditions Prim-5 whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
AB + CRISPR1-dab2 standard conditions Prim-5 whole organism dab2 expression decreased amount, abnormal
Prim-5 whole organism tcf7l1a expression increased amount, abnormal
Prim-5 whole organism tcf3b expression increased amount, abnormal
Prim-5 whole organism axin2 expression increased amount, abnormal
f2Tg/+ + CRISPR1-dab2 standard conditions Long-pec atrium cardiac muscle cell decreased amount, abnormal
Long-pec cardiac ventricle cardiac muscle cell decreased amount, abnormal
ia5Tg/+ ; zn1Tg/+ + CRISPR1-dab2 standard conditions Prim-5 forebrain mCherry expression increased amount, abnormal
Prim-5 hindbrain mCherry expression increased amount, abnormal
Prim-5 midbrain mCherry expression increased amount, abnormal
Prim-5 post-vent vasculature vasculature development decreased process quality, abnormal
Prim-5 pronephros anterior region mCherry expression increased amount, abnormal
Prim-5 vasculature posterior region mCherry expression increased amount, abnormal
w32Tg/+ ; f2Tg/+ + CRISPR1-dab2 control Long-pec cardiac muscle cell decreased amount, abnormal
Long-pec heart physical object quality, abnormal
Long-pec whole organism malformed, abnormal
w32Tg/+ ; f2Tg/+ + CRISPR1-dab2 heat shock Long-pec cardiac muscle cell amount, ameliorated
Long-pec heart physical object quality, ameliorated
Long-pec whole organism morphology, ameliorated
Acknowledgments:
ZFIN wishes to thank the journal Proceedings of the National Academy of Sciences of the United States of America for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Proc. Natl. Acad. Sci. USA