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

Expression of DDX3 and casein kinase 1ε in the hippocampus of the amyotrophic lateral sclerosis transgenic mice

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。

Objective To detect the expression of DDX3 and casein kinase 1ε（ CK1ε） in the hippocampus of the amyotrophic lateral sclerosis（ ALS） transgenic mice,and to explore the relationship between the expression of DDX3 or CK1ε and the pathogenesis of ALS. Methods Thirty-three wild ALS transgenic mice and thirty-three wild-type mice were killed at different stages. The expressions of DDX3 and CK1ε were detected by RT-PCR and Western blotting. The distribution of DDX3 and CK1ε positive cells in the hippocampus and the coexpression of DDX3 or CK1ε with astrocytes or neurons in the hippocampus were detected by a double immunofluorescence technology. Results DDX3 mRNA and protein in the hippocampus of ALS mice were of no significant differences at the early stage,but decreased at the middle and end stages,compared with wild-type mice. CK1ε mRNA and protein were all decreased at the three different stages,compared with the wild-type mice. Double immunofluorescence result showed that DDX3 and CK1ε positive cells were co-localized in the district demtate gyrusc（ DG） and CA of the hippocampus. DDX3 and CK1ε mainly expressed in neurons,not in astrocytes. Compared with the wild-type mice,the immunoreactivity of DDX3 and CK1ε in the hippocampus of ALS mice decreased obviously. Conclusion The expression of both DDX3 and CK1ε mRNA and protein decrease in the hippocampus of ALS mice,compared with the wild-type mice,indicating that the abnormal expression of DDX3 and CK1εis closely related to hippocampus lesion in ALS.