阿尔茨海默病病变相关脑区Pgc-1alpha敲除小鼠模型的构建

Construction of Pgc-1alpha knockout mouse model in Alzheimer′s disease related pathological brain regions

目的:构建在阿尔茨海默病(Alzheimer′s disease,AD)病变相关脑区过氧化物酶体增殖物激活受体γ辅激活因子-1α(peroxisome proliferator-activated receptorγcoactivator-1 alpha,Pgc-1alpha)敲除小鼠。方法:引入Pgc-1alpha条件性敲除杂合子小鼠(Pgc-1alphafl/+),通过自交获得Pgc-1alpha条件性敲除纯合子小鼠(Pgc-1alphafl/fl)。将Pgc-1alphafl/fl纯合子小鼠和在AD病变脑区(皮层、海马)特异性表达Cre重组酶的Cre工具鼠(Calb1-Cre)杂交,提取子代转基因小鼠基因组DNA,行PCR鉴定,确定其基因型;采用蛋白免疫印迹技术检测转基因小鼠与野生型小鼠脑区及肾脏PGC-1α蛋白表达。结果:双重PCR鉴定结果显示,同时扩增出400 bp及444 bp条带的小鼠为AD病变脑区Pgc-1alpha敲除纯合子小鼠(Pgc-1alphafl/fl::Calb1-Cre,Pgc-1alpha-/-)。蛋白免疫印迹结果显示,与野生型小鼠相比,条件性敲除小鼠脑区PGC-1α蛋白表达明显减少(P<0.05)。结论:成功构建在AD病变相关脑区Pgc-1alpha敲除小鼠。

Objective:To constuct peroxisome proliferator-activated receptorγcoactivator-1 alpha(Pgc-1alpha)knockout mouse model in Alzheimer′s disease(AD)-related pathologic regions.Methods:Pgc-1alphafl/fl homozygous mice were obtained by crossing Pgc-1alphafl/+heterozygous offspring.Pgc-1alphafl/fl homozygous mice were hybridized with Cre tool mice(Calb1-Cre),which specifically expressed Cre recombinase in AD lesion brain regions(cortex and hippocampus),and the genetic DNA of progeny transgenic mice was extracted and identified by PCR.The expression of PGC-1αprotein in brain and kidney of transgenic mice and wild-type mice was detected by Western blotting.Results:Dual PCR analysis revealed that mice exhibiting simultaneous amplification of 400 bp and 444 bp bands were characterized by Pgc-1alpha deletion specific to AD-related brain regions(Pgc-1alphafl/fl::Calb1-Cre,Pgc-1alpha-/-).The results of Western blotting showed that the expression of PGC-1αprotein in the brain region of conditioned knockout mice was significantly decreased compared with that of wild-type mice(P<0.05).Conclusion:Conditional Pgc-1alpha knockout mice in AD-related pathological brain regions were successfully constructed.