Abstract: In order to explore the regulatory effect of deltaD gene on vascular endothelial growth factor（VEGF） family genes vegfaa and flt4 in zebrafish（Danio rerio）,in this study, through the imaging of red fluorescence transgenic zebrafish line, we found that the defected angiogenesis of intersegmental vessels（ISVs） in deltaD homozygous mutants, and RT-qPCR experiments detected that the expression of vegfaa and flt4 of VEGF family genes were down-regulated in deltaD homozygous mutant at 1 dpf（day post fertilization）,and the expression of flt4 gene was significantly down-regulated. The sequence of deltaD, vegfaa and flt4 genes of zebrafish were obtained from genome browser database of University of California, Santa Cruz, and the expression vector p3 xFLAG-CMV-deltaD and luciferase reporter gene vectors pGL3-vegfaa-Luc and pGL3-flt4-Luc were successfully constructed. The luciferase assay showed that the activity of pGL3-vegfaa-Luc and pGL3-flt4-Luc reporter gene vectors in HEK293 T cells was about 92 times and 3.9 times higher than that of the control group, respectively. Furthermore, p3 xFLAG-CMV-deltaD was co-transfected with pGL3-vegfaa-Luc and pGL3-flt4-Luc into HEK293 T cells, comparing with the control group, the luciferase activity of p3 xFLAG-CMV-deltaD was 2.3 times and 3.7 times, respectively. This study confirmed that the ligand gene deltaD of Notch signaling pathway can play a role in vascular development by regulating vegfaa ligand and flt4 receptor genes in vascular endothelial growth factor family, which provides experimental materials and scientific basis for studying the regulatory mechanism of Notch signaling pathway in vascular development.