Background and objective: A transgenic mouse, Vegfr2-luc, in which a luciferase reporter (luc) is under control of the murine VEGFR2 promoter, can be used to track angiogenesis in vivo. The aim of this study is to identify the offspring of Vegfr2-luc transgenic mouse.
Methods: Luc was detected with PCR in genomic DNA of the new-born mouse. Luc expression in the offspring of Vegfr2-luc transgenic mouse was monitored with IVIS in vivo imaging system during post-natal development. Wound-healing models of Vegfr2-luc transgenic mouse offspring were established and the expression of luc was monitored during the wound-healing process. Luc activity and VEGFR2 mRNA expression in different organs were detected with luc Assay System and Real-time PCR respectively.
Results: PCR showed that 50% (56/112) of the offspring of Vegfr2-luc transgenic mouse carry luc. IVIS in vivo imaging results demonstrated that luc expression in Vegfr2-luc transgenic mouse dropped dramatically with age increase (P<0.001) and luc expression in the wound first increased and then decreased during the wound-healing process (P<0.001). Luc activity in female Vegfr2-luc transgenic mouse organs was positively correlated with VEGFR2 mRNA expression (r=0.948, P<0.001). Except testis, luc activity in male Vegfr2-luc transgenic mouse organs was also positively correlated with VEGFR2 mRNA expression (r=0.836, P<0.001).
Conclusion: The offspring of Vegfr2-luc transgenic mouse is applicable to tracking angiogenesis in vivo.
背景与目的: Vegfr2-luc转基因小鼠体内血管内皮生长因子受体2（vascular endothelial growth factor receptor 2, VEGFR2）的表达可以驱动荧光素酶报告基因（luciferase, luc）的表达，是活体动物水平实时监测血管生成情况的有利工具。本研究旨在对子代Vegfr2-luc转基因小鼠进行鉴定，以确定能否用于血管生成研究。
方法: PCR检测新生小鼠基因组内luc基因；利用活体成像技术观察新生Vegfr2-luc转基因小鼠生长发育过程中以及皮肤伤口修复过程中luc基因表达水平的变化情况；荧光素酶报告基因检测试剂盒检测成年（8周龄）转基因小鼠各脏器荧光素酶的活性和Real-time PCR检测各器官VEGFR2 mRNA的表达水平。
结果: PCR结果显示50%（56/112）的新生小鼠携带luc基因。活体成像结果显示随着Vegfr2-luc转基因小鼠发育成熟，luc表达量逐渐降低（P < 0.001）；在皮肤伤口修复过程中，伤口处luc表达水平先增强后降低（P < 0.001）。雌性成年转基因小鼠各脏器VEGFR2 mRNA的表达水平与荧光素酶活性呈正相关（r=0.948, P < 0.001）。将睾丸组织除外，雄性成年转基因小鼠各脏器VEGFR2 mRNA的表达水平与荧光素酶活性同样呈正相关（r=0.836, P < 0.001）。
结论: Vegfr2-luc转基因子代小鼠体内luc表达水平的变化可以反映VEGFR2的表达情况。