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ZIRC
ZFIN ID: ZDB-FIG-160203-14
Samsa et al., 2015 - Cardiac contraction activates endocardial Notch signaling to modulate chamber maturation in zebrafish. Development (Cambridge, England)   142:4080-91 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-15 to Protruding-mouth
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Long-pec to Protruding-mouth

Fig. 6

Shear stress promotes notch1 expression in a primary cilia-dependent manner. (A-B′) The hearts of Tg(actb2:Arl13b-GFP) embryos were examined by confocal microscopy to assess localization of primary cilia in the endocardium. (A) Tg(actb2:Arl13b-GFP) reporter expression in the heart. The schematic diagram indicates the orientation of the heart and confocal section relative to the whole embryo at 30hpf. (B) High-resolution view of a Tg(actb2:Arl13b-GFP) embryo merged with (B′) a bright field image demonstrating colocalization of the primary cilium base with an endocardial cell. (C,D) Whole-mount Tg(myl7:dsRed); Tg(Tp1:VenusPest) double transgenic (C) control and (D) ift88 morphants (MO) at 48hpf. The hearts are marked with dashed circles. (C′,D′) Confocal maximal projection of the heart from the individual embryos shown in C,D, overlaying cardiomyocytes (red) and Notch reporter (green). (E,F) Confocal optical section of the (E) control and (F) ift88 morphant embryo cardiomyocytes (red) at 80hpf. The insets marked by the dotted rectangle were magnified in E′,F′. (G) Quantification of ventricular Notch reporter (EGFP) MFI from whole-mount embryos at 48hpf. (H,I) Relative expression of (H) notch1b and (I) nrg1 in control and ift88 morphant hearts. (J-L) Expression of (J) Notch1, (K) Efnb2 and (L) Nrg1 in DMSO- or 50µM CBD-treated MLECs that were exposed to 2 dyn/cm2 shear stress for 4h compared with static DMSO- and CBD-treated controls. Red arrows highlight Arl13b-GFP in the endocardium. Blue arrows highlight Notch reporter signal in neural tissue. White arrows highlight trabeculae. *P≤0.05-0.01, **P≤0.01-0.001, ***P<0.001 compared with control morphants (one-sample t-test compared with 1.0-fold change or Student′s t-test). Error bars are s.e.m. Scale bars: 50µm in A; 10µm in B,B′,F&prime& 100µm in F. a, atrium; CBD, ciliobrevin D; lu, lumen; MFI, mean fluorescence intensity; v, ventricle.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
DsRed2 s940Tg ; vc6Tg control Long-pec cardiac muscle cell ISH
Protruding-mouth cardiac muscle cell ISH
s940Tg ; vc6Tg + MO1-ift88 + MO3-ift88 control Long-pec cardiac muscle cell IFL
Protruding-mouth cardiac muscle cell IFL
GFP hsc5Tg control Prim-15 endocardium cilium IFL
notch1b twu26Tg + MO1-ift88 + MO3-ift88 control Long-pec heart RTPCR
nrg1 twu26Tg + MO1-ift88 + MO3-ift88 control Long-pec heart RTPCR
Venus s940Tg ; vc6Tg control Long-pec atrioventricular canal ISH
Long-pec brain ISH
s940Tg ; vc6Tg + MO1-ift88 + MO3-ift88 control Long-pec atrioventricular canal endocardium IFL
Long-pec cardiac ventricle RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
s940Tg ; vc6Tg + MO1-ift88 + MO3-ift88 control Long-pec atrioventricular canal endocardium Venus expression decreased amount, abnormal
Long-pec cardiac ventricle Venus expression decreased amount, abnormal
Protruding-mouth trabecular layer absent, abnormal
twu26Tg + MO1-ift88 + MO3-ift88 control Long-pec heart notch1b expression decreased amount, abnormal
Long-pec heart nrg1 expression decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development