突变SOD1 cDNA在大鼠皮层神经元定位和作用

Location and Role of Mutant Cu/Zn Superoxide Dismutase in Primary Cultured Cortical Neurons of Rat

目的:分析一个ALS家系突变铜、锌超氧化物歧化酶(Cu/Zn superoxide dismutase,SOD1)基因在原代培养的大鼠皮层神经元中的定位及作用。方法:将构建好的含有SOD1正常基因与突变基因在内的增强型绿色荧光蛋白真核表达载体(pEGFP-hSOD1和pEGFP-mSOD1)分别转染大鼠皮层神经元,转染后观察绿色荧光在细胞内的分布情况,同时设对照组,检测神经元的SOD1活性、MDA含量及细胞活性。结果:转染后荧光物质表达于胞浆内,与转染pEGFP-mSOD1质粒的神经元相比,转染pEGFP-hSOD1质粒神经元的SOD1活性、MTT值显著增高,而MDA、LDH水平显著降低。结论:突变SOD1蛋白的酶活性下降,使脂质过氧化物相对增多,导致神经元损伤,可能是该家系发病的主要原因之一。

Objective:To analyze the role and location of mutant Cu/Zn superoxide dismutase(SOD1)gene from an amyotrophic lateral sclerosis(ALS)family in primary cultured cortical neurons.Methods:Plasmid-enhanced green fluorescent protein(pEGFP)including normal or mutant SOD1 gene was respectively transfected into neurons.The location of green fluorescent protein(GFP)was observed with contrast phase microscope.The activity of SOD,level of MTT and neuron activity were detected by Kits.Results:Green fluorescence was foun...