Fig. 4

Abdominal aortic aneurysm formation in zebrafish embryos following treatment with Angiotensin II or snuff extract. (A) Confocal micrographs of the central trunk region in fli1a:EGFP embryos at 4 days post-fertilization (dpf) either non-treated (control), injected at the 1-cell stage with 50ng angiotensin II (AngII) in the yolk, or treated with cigarette smoke extract (CS) at 1:30 dilution or snuff extract at 1:50 dilution from the 2dpf stage. The aorta is highlighted in a magnified image under the overview image and the abdominal (anterior) and distal (posterior) parts of the aorta further magnified in the left and right images, respectively, under the isolated aorta image. Red dots outline the aortic wall. Scale bars: 100 µm, 50µm and 50µm in the overview, aorta highlight and posterior aorta highlight images, respectively. (B) Quantification of the average aorta diameter in the abdominal region (anterior aorta images shown in A) of control zebrafish embryos during development. n=20, 9, 20, 19 and 12 embryos at 24, 48, 72, 98 and 120hpf, respectively. *P<0.05, ***P<0.001. (C) Quantification of the average aorta diameter in the abdominal region (anterior aorta images shown in A) of zebrafish embryos treated with CS, snuff or AngII as in A. n=19, 7, 7 and 13 in the control, CS, Sn and AngII groups respectively. **P<0.01, ***P<0.001. All experiments were repeated twice with similar results. Results are shown as means±s.e.m. and statistical evaluation was done using Student’s two-tailed t-test assuming equal variance between the groups.