Fig. 3

Block of Mevalonate Pathway Causes Cardiovascular Failure in Zebrafish Embryos by Reducing CoQ10 Synthesis(A) Wild-type (WT) embryos at 72 hpf treated from 54 hpf with statin (mevastatin) or DMSO. Statin treatments induce a bar-like phenotype which is characterized by hemorrhages (asterisk) and heart failure (arrowhead). Scale bar, 300 µm.(B) Three dimensional projections of trunk vessels at 72 hpf of DMSO and statin-treated zebrafish embryos. Statin treatments induce specific endothelial vessels regression and fragmentation in DA and Se (arrows). Scale bar, 100 µm.(C) Quantification of bar-like phenotype after statin treatments. MEV, mevastatin 500 nM; SIM, simvastatin 500 nM; MEN, mevinolin 500 nM.(D) and cholesterol (E) detected by HPLC-UV analyses in statin-treated embryos.(F and G) Bright-field (F) and fluorescent (G) images of Tg(kdrl:GFP)^s843ubiad1^+/+ and ubiad1^+/- embryos at 65 hpf treated with mevastatin (2.5 nM and 5 nM) from 32 to 50 hpf. Treatment with a low dose of statin caused hemorrhages (asterisks) and altered heart morphology (arrowheads) in ubiad1^+/-, but not in ubiad1^+/+ embryos. Although treatment with a low dose of statin did not induce specific alterations of the main vasculature, ubiad1^+/- embryos showed head vasculature, Se, and DA regression; scale bar, 300 µm.(H) Images of Tg(kdrl:GFP)^s843ubiad1^+/+ and ubiad1^+/- embryos trunk vasculature show thinner and collapsed Se and DA morphology in statin-treated ubiad1^+/- but not in ubiad1^+/+ embryos. Scale bar, 75 µm.(I) Penetrance of bar-like phenotype at 65 hpf after treatment with mevastatin. ubiad1^+/- embryos are significantly more sensitive to develop a bar-like phenotype than normal embryos (ubiad1^+/+).(J) CoQ10 supplemented embryos show less susceptibility to statin treatments in terms of developing a bar-like phenotype as indicated by histograms showing the percentage of statin-treated embryos having bar-like phenotype after exogenous CoQ10 delivery.(K) CoQ10 supplementation before statin treatment prevents cardiac edema (arrowhead) and brain hemorrhages (asterisk) in barolo. Images of vehicle- and CoQ10-injected embryos after statin treatment. Scale bar, 300 µm.(L) Histograms show oxidative stress in embryos after statin treatment with or without CoQ10.All data are means ± SEM. p < 0.001.See also Figure S3.