转录因子12在SOD1-G93A转基因小鼠大脑皮层和纹状体中的表达与定位研究

The expression and localization of transcript factor 12 in cerebral cortex and striatum of SOD1-G93A transgenic mice

目的:检测转录因子12(TCF12)在SOD1-G93A转基因小鼠不同年龄阶段大脑皮层和纹状体中的表达情况。方法:采用成年SOD1-G93A转基因小鼠和野生型(WT)小鼠,分别在转基因小鼠肌萎缩性侧索硬化症(ALS)发病的早(95 d)、中(108 d)、晚(122 d)期取材制备标本。使用real time RT-PCR技术检测TCF12的mRNA表达情况,使用Western Blot技术检测TCF12蛋白表达情况,使用免疫荧光双标记技术检测TCF12在大脑皮层和纹状体的表达分布以及细胞类型分布。结果:在SOD1-G93A转基因小鼠和WT小鼠的大脑皮层和纹状体中均检测到了TCF12分布,TCF12在神经元表达;在ALS发病早、中、晚期SOD1-G93A转基因小鼠大脑皮层中的TCF12 mRNA和蛋白表达较WT小鼠显著升高(P<0.05,P<0.01);在发病中、晚期SOD1-G93A转基因小鼠纹状体中的TCF12 mRNA和蛋白水平表达相较于WT小鼠显著升高(P<0.01)。结论:TCF12在SOD1-G93A转基因小鼠发病中晚期大脑皮层和纹状体中表达升高,提示TCF12分子参与大鼠了ALS发病中、晚期神经元的退变进程。

Objective:To detect the expression of transcript factor 12(TCF12)in cerebral cortex and striatum of amyotrophic lateral sclerosis(ALS)transgenic mice at different ages.Methods:We selected adult SOD1-G93 A transgenic mice and wild-type(WT)mice to prepare specimens on early stage(95 d),middle stage(108 d),and end stage(122 d).Real-time quantitative PCR was used to detect the expression of TCF12 mRNA,Western Blot was used to detect the expression of TCF12 protein,and immunofluorescence double labeling technique was used to observe the distribution of TCF12 positive cells and cell types in cerebral cortex and striatum.Results:TCF12 positive cells were found in cerebral cortex and striatum of both SOD1-G93 A transgenic mice and WT mice,and TCF12 expressed in neurons.TCF12 expressions in cerebral cortex of ALS mice were higher than those of WT mice in the early,middle and end stages,and the differences were statistically significant(P<0.05,P<0.01);TCF12 expressions in the striatum of SOD1-G93 A transgenic mice in the middle and end stages were significantly higher than those of WT mice(P<0.01).Conclusion:The increased expressions of TCF12 were found in the cerebral cortex and striatum of SOD1-G93 A transgenic mice at middle and end stages,suggesting that TCF12 was involved in the neuron degeneration of ALS in the middle and end stages.