ZFIN ID: ZDB-FIG-170302-15
Chen et al., 2017 - Cilia Control Vascular Mural Cell Recruitment in Vertebrates. Cell Reports   18:1033-1047 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Terms:
Stage Range: Long-pec to Protruding-mouth
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: Long-pec to Protruding-mouth

Fig. 5

Cilia Are Required to Drive vMC Recruitment

(A) Pharmacological impairment of primary cilia formation negatively regulates vMC recruitment. Confocal images of partial z-projections of the trunk region (somite 8–14) of Tg(kdrl:egfp)s843;Tg(acta2:mCherry)uto5 embryos at 80 hpf (merged and single channels) after disrupting cilia formation by CBD treatment. Compared to controls, CBD-treated embryos show reduced vMC coverage around the DA (arrows). Scatter plots show the quantification of vMC number per 500 μm of the DA at 80 hpf. n = 39 and 53 embryos. Data are represented as mean ± SD. Stars represent the results of unpaired t tests of mean difference = 0 (p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001). g, gut.

(B) Endothelial-specific genetic inhibition of ift88 gene expression impairs vMC coverage. Confocal images of partial z-projections of the trunk region (somite 8–14) of Tg(kdrl:egfp)s843Tg(acta2:mCherry)uto5 embryos at 80 hpf (merged and single channels) injected with the integratable CRISPR vector for endothelial-specific ift88 gene targeting. Cmlc2:GFP-negative embryos were used a negative control (ctrl). Scatter plots show the quantification of vMC number per 500 μm of the DA at 80 hpf. n = 20 and 20 embryos. Data are represented as mean ± SD. Stars represent the results of unpaired t tests of mean difference = 0 (p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001).

(C) Inhibition of cilia function by CBD treatment impairs Notch signaling in living zebrafish blood vessels. qPCR analysis of Notch target genes in DMSO and 1 and 2 μM CBD-treated embryos. qPCR analysis of Notch target gene expression in zebrafish treated with different concentrations of CBD at 48 hpf. A zebrafish trunk was used for RNA extraction and qPCR analyses. Notch1b, dll4, and Jagged1b are the most affected Notch targets by the lack of cilia at this stage of development (48 hpf). Data are represented as mean ± SD. Stars represent the results of two-way ANOVA. p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001.

See also Figures S7–S10.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
dll4 WT chemical treatment by injection: ciliobrevin D Long-pec trunk RTPCR
EGFP s843Tg; uto5Tg control Protruding-mouth dorsal aorta IFL
Protruding-mouth intersegmental vessel IFL
Protruding-mouth posterior cardinal vein IFL
s843Tg; uto5Tg chemical treatment by injection: ciliobrevin D Protruding-mouth dorsal aorta IFL
Protruding-mouth intersegmental vessel IFL
Protruding-mouth posterior cardinal vein IFL
hey1 WT chemical treatment by injection: ciliobrevin D Long-pec trunk RTPCR
hey2 WT chemical treatment by injection: ciliobrevin D Long-pec trunk RTPCR
jag1b WT chemical treatment by injection: ciliobrevin D Long-pec trunk RTPCR
mCherry s843Tg; uto5Tg control Protruding-mouth dorsal aorta vascular associated smooth muscle cell IFL
Protruding-mouth gut IFL
s843Tg; uto5Tg chemical treatment by injection: ciliobrevin D Protruding-mouth dorsal aorta anatomical region IFL
Protruding-mouth dorsal aorta vascular associated smooth muscle cell IFL
Protruding-mouth gut IFL
notch1b WT chemical treatment by injection: ciliobrevin D Long-pec trunk RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT chemical treatment by injection: ciliobrevin D Long-pec trunk dll4 expression decreased amount, abnormal
Long-pec trunk hey1 expression decreased amount, abnormal
Long-pec trunk hey2 expression decreased amount, abnormal
Long-pec trunk jag1b expression decreased amount, abnormal
Long-pec trunk notch1b expression decreased amount, abnormal
s843Tg; uto5Tg chemical treatment by injection: ciliobrevin D Protruding-mouth dorsal aorta anatomical region mCherry expression decreased distribution, abnormal
Protruding-mouth dorsal aorta vascular associated smooth muscle cell decreased amount, abnormal
Acknowledgments:
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