摘要
目的:观察Notch1在肌萎缩侧索硬化症(ALS)转基因鼠动物模型和细胞模型中的表达情况。方法:应用免疫荧光、免疫印迹、RT-PCR,检测Notch1在95、108、122 d ALS转基因鼠脊髓中的表达变化;检测转染pEGFP-wt-SOD1和pEGFPG93A-SOD1的NSC34细胞模型中Notch1的表达变化。结果:Notch1可与β-tubulinⅢ共表达,与GFAP无明显共表达。较同窝野生型鼠,Notch1于蛋白水平和mRNA水平上的表达在95 d ALS转基因鼠脊髓中无明显变化,在108 d和122 d ALS转基因鼠脊髓中明显升高;与转染pEGFP-wt-SOD的NSC34细胞相比,转染pEGFP-G93A-SOD1的NSC34细胞中Notch1蛋白和mRNA表达增多。结论:Notch1在ALS转基因鼠动物模型和细胞模型中表达增多,提示Notch1信号通路可能与ALS相关。
Objective： To observe the expression of Notchl in the amyotrophic lateral sclerosis（ALS） transgenic animal model and NSC34 motor neuron cellular model. Methods： The expression of Notehl in the spinal cord of ALS transgenic mice was detected by immunofluorescence,Western blotting and RT-PCR at 95,108 and 122 d respectively. The expression of Notehl in NSC34 cells transfected with pEGFP-wt-SOD1 and pEGFP-G93A-SOD1 was detected by Western blotting and RT-PCR. Results： Immunofluorescence indicated that Notch1 and β-tubulin Ⅲ eoexpressed but Notchl and GFAP didn＇t obviously. Western blotting showed that the expression of Notch1 protein was significantly increased in ALS transgenic mice compared with that in wild-type mice. Consistently, the mRNA of Notch1 detected by RT-PCR in ALS transgenic mice was also significantly higher than that in wild-type mice at 108 d and 122 d. There was no significant change in the expression of Notch1 at both protein and mRNA level at 95 d; the expression of Notch1 in NSC34 cells transfeeted with pEGFP-G93A- SOD1 was significantly higher compared with NSC34 cells transfected with pEGFP-wt-SOD1 at both protein and mRNA level. Conclusion： Notchl up-regulation in the spinal cord of the ALS transgenic mice model and cellular model suggest that the Notchl signaling pathway may be associated with ALS.
作者
杜红梅
管英俊
于丽
陈燕春
周风华
张皓云
接琳琳
刘永新
Du Hongmei Guan Yingjun Yu Li Chen Yanchun Zhou Fenghua Zhang Haoyun Jie Linlin Liu Yongxin(Department of Histology and Embryology Department of Pathology Department of Basic Medicine, Weifang Medical University, Weifang 261053, China)
出处
《解剖学杂志》
CAS
CSCD
北大核心
2017年第1期21-25,共5页
Chinese Journal of Anatomy
基金
山东省高等学校科技计划项目(J13LK05)
国家自然科学基金(81271413
81401066)
