Fig. 1

Fig. 1

Fig. 1. The effects of MNU on angiogenesis of zebrafish. A. Research protocol. At 6 hpf (early gastrula period), Tg(kdrl:eGFP)^s843 zebrafish embryos were treated with MNU solution for 1 h. Imaging analysis was performed after 48 hpf. MNU, N-methyl-N-nitrosourea. B. Different concentrations of MNU solution were used for embryos treatment at 6 hpf for 1 h. The survival rate (top), malformation rate (middle), and ectopic branching rate of blood vessels (bottom) were statistically analyzed at 24 hpf, 48 hpf, 72 hpf, 96 hpf and 120 hpf, respectively. Results for each concentration were obtained from three independent experiments. MNU of 0, 25, 50, 100 and 150 mg/L were displayed in black, gray, green, red, and blue, respectively. Data are shown as the mean ± SEM. * * P < 0.01 and * ** P < 0.001. C. Imaging analysis of the intersegmental blood vessel (ISV) ectopic budding phenotype in the control and 100 mg/L MNU treated groups. White arrows indicate ISV ectopic budding. ISV, intersegmental blood vessel. Scale bar, 50 µm. D. The percentage of embryos with ISV ectopic sprouting was calculated. Data are shown as the mean ± SEM. * ** P < 0.001. E. Representative images from control group and 100 mg/L MNU-treated groups at 48, 72 and 120 hpf (top), and representative images from brain and retinal vessels in 100 mg/L MNU treated groups at 72 hpf (bottom). Obvious ectopic vessels sprouts formed in the ISV (white arrows), brain and retina in 100 mg/L MNU treated groups. Red short arrows indicate the mesencephalic vein (MSV). Red asterisks indicate the middle cerebral artery (MCtA). Red arrowheads indicate the dorsal longitudinal vein (DLV). Red long arrows indicate the posterior cerebral vein (PCev) and purple arrows indicate the dorsal retinal vessel (DRV). Scale bars, 100 µm.