DDX3和CK1ε在SOD1-G93A突变的ALS转基因小鼠纹状体的表达

The expression of DDX3 and CK1ε in the striatum of SOD1-G93A mutate ALS mice

目的:检测DDX3和CK1ε在SOD1-G93A突变的ALS转基因小鼠纹状体的表达变化,探讨其在ALS纹状体病变中的作用,为ALS研究提供新的实验依据。方法:将发病早期、中期和晚期(即95 d、108 d和122 d)的成年野生型小鼠和SOD1-G93A突变型ALS转基因小鼠分别处死取材,运用免疫荧光技术对小鼠纹状体内DDX3和CK1ε的表达水平进行检测,运用RT-PCR技术对小鼠纹状体内DDX3和CK1ε的m RNA水平进行检测,运用Western Blot行蛋白水平变化检测。结果:野生型小鼠和SOD1-G93A突变型转基因小鼠纹状体中均可检测到DDX3和CK1ε阳性细胞。DDX3和CK1ε在神经元表达,但在星形胶质细胞不表达。在发病的中期和晚期,DDX3在ALS转基因小鼠纹状体中的m RNA和蛋白表达与野生型鼠相比均升高。CK1εm RNA在ALS发病的早期不变化,在中期、晚期较野生型鼠降低;而在ALS发病的早期转基因小鼠纹状体中CK1ε蛋白表达量较野生型鼠升高,在中期、晚期降低。结论:DDX3和CK1ε在SOD1-G93A突变的ALS转基因小鼠纹状体中表达异常与ALS纹状体的病变密切相关。

Objective： To explore the role of DDX3 and CK1ε in the pathogenesis of ALS by detecting the expression of DDX3 and CK1ε in the striatum of SOD1-G93 A ALS transgenic mice and provide a new experimental basis for ALS research. Methods： Adult wild-type mice and SOD1-G93 A mutant ALS transgenic mice were sacrificed at the early,middle,and late stages（ 95 d,108 d and 122 d）. We detected the expression pattern of DDX3 and CK1ε in the striatum of the mice by immunofluorescence technology,the m RNA levels by RT-PCR technology,and the protein expression by Western Blot technology. Results： DDX3 and CK1ε positive cells were all detected in the striatum of wild-type mice and SOD1-G93 A mutant ALS transgenic mice. DDX3 and CK1ε were expressed in neurons and not in astrocytes.In the middle and late stage,compared with the wild-type mice,the protein and m RNA expression of DDX3 in the striatum of ALS transgenic mice increased statistically. CK1ε m RNA level in the striatum of ALS transgenic mice did not change in the early stage,but decreased statistically in the middle and late stage. However,CK1ε protein level in the striatum of ALS transgenic mice increased in the early stage,but decreased statistically in the middle and late stage.Conclusion： The expression changes of DDX3 and CK1ε in the striatum of SOD1-G93 A mutant ALS transgenic mice were closely related with the striatum lesion of ALS.