DDX3和酪蛋白激酶1ε在肌萎缩侧索硬化症转基因鼠海马中的表达

发布时间：2018-08-27 11:05

【摘要】：目的检测DDX3和酪蛋白激酶1ε(CK1ε)在肌萎缩侧索硬化症(ALS)转基因鼠海马中的表达变化,揭示DDX3和CK1ε的表达改变与ALS发病的关系。方法 33只ALS转基因鼠和33只野生型鼠分别于发病不同时期取材,应用RT-PCR和Western blotting技术检测海马组织中DDX3和CK1ε的表达变化;通过免疫荧光双标染色技术观察DDX3和CK1ε阳性细胞的分布特点及其与神经元、星形胶质细胞等的共表达情况。结果与野生型鼠相比,ALS转基因鼠海马中DDX3 mRNA和蛋白在发病早期变化不明显,中期和晚期表达均明显降低;CK1εmRNA和蛋白则在发病早期、中期和晚期表达均降低。免疫荧光双标结果显示,在ALS鼠和野生型鼠海马齿状回区和CA区均可检测到DDX3和CK1ε阳性细胞。DDX3和CK1ε主要表达在神经元,在星形胶质细胞未见表达。与野生型鼠比较,ALS转基因鼠海马中DDX3和CK1ε免疫反应性明显降低。结论 DDX3和CK1εmRNA和蛋白在ALS转基因鼠海马中表达均降低,表明DDX3和CK1ε表达异常与ALS海马区病变密切相关。
[Abstract]:Objective to investigate the expression of DDX3 and casein kinase 1 蔚 (CK1 蔚) in hippocampus of (ALS) transgenic rats with amyotrophic lateral sclerosis (ALS), and to explore the relationship between the expression of DDX3 and CK1 蔚 and the pathogenesis of ALS. Methods 33 ALS transgenic mice and 33 wild type mice were collected at different stages of the disease. The expression of DDX3 and CK1 蔚 in hippocampus were detected by RT-PCR and Western blotting techniques. The distribution of DDX3 and CK1 蔚 positive cells and their coexpression with neurons and astrocytes were observed by double immunofluorescence staining. Results compared with wild-type mice, the expression of DDX3 mRNA and protein in hippocampus of ALS transgenic mice did not change significantly in the early stage, while the expression of CK1 蔚 mRNA and protein decreased significantly in the middle and late stages of the disease, while the expression of CK1 蔚 mRNA and protein decreased in the early, middle and late stages of the disease. The results of immunofluorescence double labeling showed that DDX3 and CK1 蔚 positive cells, DDX3 and CK1 蔚 were mainly expressed in neurons, but not in astrocytes, in the dentate gyrus and CA regions of ALS mice and wild type mice. Compared with wild-type mice, the DDX3 and CK1 蔚 immunoreactivity in hippocampus of ALS transgenic mice decreased significantly. Conclusion the expression of DDX3 and CK1 蔚 mRNA and protein in the hippocampus of ALS transgenic mice is decreased, which indicates that the abnormal expression of DDX3 and CK1 蔚 is closely related to the ALS hippocampal lesion.
【作者单位】：潍坊医学院组织学与胚胎学教研室;潍坊医学院人体解剖学教研室;潍坊医学院附属医院骨科;
【基金】：国家自然科学基金(81401066,81271413) 山东省自然科学基金(ZR2012HQ021,2012HL42) 山东省科技发展计划项目(2012GSF11827) 山东省教育厅课题(J11LF16,J12LK01) 潍坊医学院大学生科技创新基金(KX2016009,KX2016015)
【分类号】：R744.8

【参考文献】