DDX3和CK1ε在ALS转基因小鼠大脑皮层中的表达变化  

The changes in the expression of DDX3 and CK1ε in the cerebral cortex of ALS transgenic mice

作　　者：王箐[1] 原萌[2] 刘焕彩[3] 梁婵婵 林宝勇 张雅雯[2] 周风华[4] 陈燕春[4] 

机构地区：[1]潍坊医学院人体解剖学教研室神经疾病与再生修复重点实验室,潍坊261053 [2]潍坊医学院,临床医学专业潍坊261053 [3]潍坊医学院附属医院,潍坊261053 [4]潍坊医学院组织学与胚胎学教研室,潍坊261053

出　　处：《神经解剖学杂志》2018年第1期15-20,共6页Chinese Journal of Neuroanatomy

基　　金：国家自然科学基金(81401066),山东省自然科学基金(ZR2012HQ021;ZR2016HM60;ZR2016HL20),山东省教育厅课题(J11LF16),山东省医药卫生科技发展计划项目(2016WS0691,2016WS0666),潍坊医学院大学生科技创新基金(KX2017008,KX2016009)

摘　　要：目的:观察RNA解旋酶DDX3和酪蛋白激酶1ε(CK1ε)在ALS转基因小鼠大脑皮层组织中的表达变化,探讨DDX3与CK1ε表达改变在ALS发病中的作用和意义。方法:选择SOD1-G93A转基因小鼠进行试验,分别于发病早期(95 d)、中期(108 d)、晚期(122 d)取材。应用免疫荧光双标染色方法检测DDX3和CK1ε在ALS转基因小鼠大脑皮层中的表达变化规律及其与神经元的共定位关系。应用RT-PCR和Western Blot技术检测ALS转基因小鼠大脑皮层组织中DDX3和CK1ε在mRNA、蛋白水平的表达变化,每组均选择同年龄的野生型小鼠作为对照组。结果:与野生型鼠相比,ALS转基因小鼠大脑皮层组织中的DDX3蛋白和mRNA早期表达均无明显变化,中、晚期表达均升高;CK1ε蛋白在ALS小鼠发病早期没有明显改变,中、晚期表达升高,mRNA表达无明显变化。免疫荧光双标结果显示,ALS转基因小鼠和野生型小鼠大脑皮层组织中均可检测到DDX3和CK1ε阳性细胞,且与神经元共表达。ALS转基因小鼠大脑皮层组织内DDX3与CK1ε免疫反应性较同龄野生型鼠明显增强。结论:DDX3与CK1ε表达异常与ALS大脑皮层病变密切相关,参与了ALS发病。Objective： To investigate the expression changes of RNA helicase DDX3 and casein kinase 1 epsilon （CKls） in the cerebral cortex of ALS transgenic mice. Methods： SOD1-G93A transgenic mice were selected and tested in early, middle and late stage （95 d, 108 d and 122 d） separately. The mRNA and protein levels of DDX3 and CK1ε in the cerebral cortex of ALS transgenic mice were detected by RT-PCR and Western Blot respectively. The expression of DDX3 and CK1ε was also detected by double immunofluorescence technique to investigate the regulation of expression variation and their co-localization with neurons. Wild-type mice of the same age were selected as control group. Results： Compared with wild-type mice, the expression of DDX3 protein and mRNA in the cerebral cortex of ALS transgenic mice did not change significantly in early stage, but increased in both middle and late stage. CKI~ protein was not significantly changed in the early stage, but increased in the middle and late stage. There was no significant change in CK1ε mRNA expression. Double immunofluorescence results showed that DDX3 and CK1ε positive ceUs were detected in the cerebral cortex of ALS transgenic mice and wild-type mice and co-expressed with neurons. The immunoreactivity of DDX3 and CKle of ALS transgenic mice was enhanced compared with wild-type mice of the same age. Conclusion： The abnormal expression of DDX3 and CK1ε was closely related to ALS cerebral cortex lesion, which was involved in the pathogenesis of ALS.

关 键 词：肌萎缩侧索硬化症 转基因小鼠 大脑皮层 RNA解旋酶DDX3 酪蛋白激酶1ε 小鼠 

分 类 号：R744.8[医药卫生—神经病学与精神病学]