Neurogenin 1在肌萎缩脊髓侧索硬化症转基因鼠脊髓中的表达变化

Expression change of neurogenin 1 in the spinal cord of ALS transgenic mice

目的：研究neurogenin 1 （Ngn1）转录因子在成年肌萎缩脊髓侧索硬化症（ALS）转基因模型鼠脊髓内的表达变化,探讨其在ALS转基因鼠发病中的作用.方法：选取ALS转基因鼠和同窝野生型鼠各18只,分别于95、108 d和122 d取材,部分鼠经4％多聚甲醛心灌注固定,分离脊髓、制备冷冻切片;部分鼠分离脊髓、提取蛋白和RNA,分别应用免疫荧光显色、免疫印迹和RT-PCR检测Ngnl蛋白及mRNA在ALS转基因鼠发病中的变化.结果：与同窝野生型鼠比较,ALS转基因鼠Ngnl mRNA和蛋白表达在95、108 d和122 d均降低.免疫荧光实验结果显示,Ngnl阳性细胞主要分布在脊髓灰质前角,与同窝野生型鼠比较,ALS转基因鼠Ngnl阳性细胞光密度值明显减小.结论：在ALS转基因鼠发病脊髓中Ngnl阳性细胞光密度值明显减少,mRNA和蛋白表达降低,表明Ngnl与ALS的发生密切相关.

Objective： To study the expression of neurogenin 1 （Ngnl） transcription factor in the spinal cord of adult amyotrophic lateral sclerosis （ALS） transgenic mice, and to explore the mechanism of Ngnl transcription factor in the pathogenesis of ALS transgenic mice. Methods： Both the ALS transgenic mice and wild-type mice were selected, and some were perfused intracardially with 4% paraformaldehyde at the age of 95 d, 108 d and 122 d. Their spinal cords were separated and sectioned. Some were anesthetized and spinal cords were quickly removed. RNA and total protein were obtained and observed by immunofluorescence technique, Western blotting and RT-PCR. Results： Compared with wild-type mice, the expression of Ngnl mRNA and total protein levels were reduced at 95 d, 108 d and 122 d in the ALS transgenic mice. Immunofluorescence results showed that Ngnl positive cells were reduced in spinal cord of ALS transgenic mice at different ages compared with litter born-wild type mice. Conclusion： Ngnl positive cells and the expression of Ngnl mRNA and protein were both reduced in the spinal cord of ALS transgenic mice, suggesting that Ngnl was closely related to the pathogenesis of amyotrophic lateral sclerosis.