Tnfrsf11a^(Cre)介导黄色荧光素蛋白有效标记脑小胶质细胞

Tnfrsf11a^(Cre)mediates YFP labeling some of tissue macrophages

目的构建Tnfrsf11a^(Cre)Rosa26^(yfp)报告基因小鼠,检测Tnfrsf11a^(Cre)介导黄色荧光素蛋白YFP标记组织巨噬细胞的效率。方法将Tnfrsf11a^(Cre)小鼠与Rosa26^(yfp)小鼠交配,通过PCR筛选出Tnfrsf11a^(Cre)Rosa26^(yfp)报告基因小鼠。分离成年Tnfrsf11a^(Cre)Rosa26^(yfp)小鼠脑小胶质细胞、肝脏巨噬细胞、肾脏巨噬细胞、肺泡巨噬细胞、脾脏巨噬细胞,标记流式抗体,通过流式细胞术分析Tnfrsf11a^(Cre)介导荧光素蛋白YFP标记组织巨噬细胞的效率。结果Tnfrsf11a^(Cre)介导黄色荧光素蛋白YFP对脑小胶质细胞具有约91.27%的标记效率,但对肝脏、脾脏、肺泡巨噬细胞细胞的标记效率分别只有约63.60%、69.66%、32.76%。结论Tnfrsf11a^(Cre)可以介导YFP对脑小胶质细胞进行示踪。同时,Tnfrsf11a^(Cre)可用做脑小胶质细胞基因条件性敲除的工具鼠。

Objective Tnfrsf11a^(Cre)Rosa26^(yfp)reporter gene mice were prepared to determine the efficiency of Cre-mediated recombination using flow cytometry in different tissue-resident macrophages.Methods The Tnfrsf11a^(Cre)Rosa26^(yfp)reporter mice were generated by crossing Tnfrsf11a^(Cre)mice with Rosa26^(yfp)mice and identified by PCR.Brain microglia,liver macrophages,kidney macrophages,alveolar macrophages and spleen macrophages were separated from adult Tnfrsf11a^(Cre)Rosa26^(yfp)reporter mouse and yellow fluorescent protein(YFP)labeling efficiency was analyzed by flow cytometry.Results YFP expression percentage was about 91.27%of brain microglia in Tnfrsf11a^(Cre)Rosa26^(yfp)reporter mice,but liver,spleen,and alveolar macrophages were inefficiently targeted(63.60%,69.66%,32.76%).Conclusion Tnfrsf11a^(Cre)mice were qualified as conditional knockout model mice of brain microglia and Tnfrsf11a^(Cre)Rosa26^(yfp)reporter mice can be used as a tool for conditional knockout of brain microgliain vivo.