Transcription profiling of mouse laser capture microdissected SOD1 G93A motor neurons reveals cellular pathways involved in the adaptation and progression of motor neuron injury in the mouse model of familial ALS

Microarray analysis has been applied to the study of ALS in order to investigate gene expression in whole spinal cord homogenates of SOD1 G93A mice and human ALS cases, although the massive presence of glial cells and inflammatory factors has made it difficult to define which gene expression changes were motor neuron specific. Recently, laser capture microdissection (LCM), combined with microarray analysis, has allowed the identification of motor neuron specific changes in gene expression in human ALS cases. The aim of the present study is to combine LCM and microarray analysis to study how motor neurons in the spinal cord of transgenic SOD1 G93A mice and transgenic SOD1 WT respond to stimuli determined by the presence of the human mutant protein throughout the evolution of the stages in motor neuron injury Experiment Overall Design: Motor neurons have been isolated from the spinal cord of G93A mice and non transgenic littermates at different time points and the transcription expression profile of the isolated motor neurons has been analysed为探究SOD1 G93A小鼠全脊髓匀浆及人类肌萎缩侧索硬化（Amyotrophic Lateral Sclerosis, ALS）样本中的基因表达水平，本研究此前已应用基因芯片分析（microarray analysis）开展ALS相关研究，但由于胶质细胞与炎症因子大量富集，难以明确哪些基因表达变化为运动神经元特异性改变。 近期，激光捕获显微切割（Laser Capture Microdissection, LCM）联合基因芯片分析技术，已实现在人类ALS患者样本中鉴定运动神经元特异性的基因表达变化。 本研究的核心目标为联合激光捕获显微切割与基因芯片分析技术，探究在运动神经元损伤的整个病程阶段中，转基因SOD1 G93A小鼠及转基因SOD1野生型（Wild Type, WT）小鼠脊髓内的运动神经元，如何响应由人类突变蛋白存在所引发的刺激信号。 实验整体设计： 本实验将在不同时间点从G93A小鼠及非转基因同窝仔鼠的脊髓中分离运动神经元，并对分离得到的运动神经元的转录表达谱进行分析。