Zhou Fenghua, Guan Yingjun, Chen Yanchun, Zhang Caixia, Yu Li, Gao Hailing, Du Hongmei, Liu Bing, Wang Xin
Department of Pathology, Weifang Medical University Weifang, Shandong, P. R. China.
Int J Clin Exp Pathol. 2013 Aug 15;6(9):1826-38. eCollection 2013.
The pathogenesis of amyotrophic lateral sclerosis (ALS) remains unclear. Accumulating evidence indicates that various miRNAs expressed in a spatially and temporally controlled manner in the nervous system have an important function in the development of neurodegenerative diseases. The present study aimed to determine the expression and cellular distribution of miRNA-9 in the spinal cord of G93A-SOD1 mutant mice at different time points (post-natal 95, 108 and 122 d). miRNA expression was evaluated by microarray analysis; differentially expressed miRNAs were validated by RT-qPCR. The cellular distribution of miRNA-9 was analyzed by in-situ hybridization. Microarray results indicated for the first time that various miRNAs were differentially expressed between the G93A-SOD1 mutant mice and the littermate control mice. miRNA-9 expression was upregulated at 95, 108, and 122 d as validated by microarray analysis, RT-qPCR, and ISH. ISH results also showed that the miRNA-9-positive cells mainly expressed in the cytoplasm were located in the dorsal horn and the ventral horn of the spinal cord. The majority of miRNA-9-positive cells were located in the ventral horn of the gray matter, the locus of neurodegeneration. These results indicated that the differential expression of miRNA-9 may have an important function in the pathogenesis of G93A-SOD1 transgenic mice.
肌萎缩侧索硬化症（ALS）的发病机制仍不清楚。越来越多的证据表明，在神经系统中以时空控制方式表达的各种微小RNA（miRNA）在神经退行性疾病的发展中具有重要作用。本研究旨在确定miRNA-9在G93A-SOD1突变小鼠脊髓中不同时间点（出生后95、108和122天）的表达及细胞分布。通过微阵列分析评估miRNA表达；通过逆转录定量聚合酶链反应（RT-qPCR）验证差异表达的miRNA。通过原位杂交分析miRNA-9的细胞分布。微阵列结果首次表明，G93A-SOD1突变小鼠和同窝对照小鼠之间各种miRNA存在差异表达。经微阵列分析、RT-qPCR和原位杂交验证，miRNA-9在95、108和122天表达上调。原位杂交结果还显示，主要在细胞质中表达的miRNA-9阳性细胞位于脊髓背角和腹角。大多数miRNA-9阳性细胞位于灰质腹角，即神经退行性变的部位。这些结果表明，miRNA-9的差异表达可能在G93A-SOD1转基因小鼠的发病机制中具有重要作用。
