DDX3和酪蛋白激酶1&epsilon;在肌萎缩侧索硬化症转基因鼠脑干中的表达变化

张雅雯 王箐 原萌 刘焕彩 王巧真 丁昊宇 周风华 陈燕春*

doi:10.16098/j.issn.0529-1356.2018.01.001

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解剖学报 ›› 2018, Vol. 49 ›› Issue (1) : 1-6. DOI: 10.16098/j.issn.0529-1356.2018.01.001

神经生物学

DDX3和酪蛋白激酶1ε在肌萎缩侧索硬化症转基因鼠脑干中的表达变化

张雅雯^1,2 王箐³原萌² 刘焕彩⁴王巧真³ 丁昊宇²周风华²陈燕春^2*

作者信息 +

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

ZHANG Ya-wen ^1,2 WANG Qing³YUAN Meng² LIU Huan-cai⁴WANG Qiao-zhen³ DING Hao-yu² ZHOU Feng-hua² CHEN Yan-chun ^2*

Author information +

文章历史 +

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)转基因鼠脑干中的表达变化，探讨DDX3和CK1ε在ALS脑干运动神经元变性中的作用。
方法选取ALS转基因鼠33只，分别于发病早期（95d）、中期（108d）和晚期（122d）3个时间点剥离脑干，应用RT-PCR、Western blotting和免疫荧光染色技术分别检测ALS转基因鼠脑干组织中DDX3和CK1ε的表达规律，在脑干运动核团舌下神经（12N）和面神经（7N）中阳性细胞的分布特点及细胞定位，每组均选择相同数量的同窝野生型鼠作为对照。结果 RT-PCR和Western blotting结果显示，与同窝野生型鼠相比，ALS转基因鼠脑干组织中DDX3和CK1εmRNA于95 d、108d、122d表达均无明显变化，DDX3和CK1ε蛋白在95d和108d表达上调，122d表达下调(P<0.01, P<0.001)。免疫荧光结果显示，在ALS鼠和野生型鼠脑干的12N和7N区域均可检测到DDX3和CK1ε阳性细胞，DDX3和CK1ε表达在神经元，在星形胶质细胞不表达。ALS鼠和野生型鼠DDX3和CK1ε免疫反应性具有差异。结论 DDX3和CK1ε在脑干中的表达异常与ALS发病密切相关。

Abstract

Objective To detect the expression of DDX3 and casein kinase 1ε (CK1ε）in the brain stem of amyotrophic lateral sclerosis (ALS) transgenic mice and study the role of DDX3 and CK1ε in the degeneration of brain stem motor neurons of amyotrophic lateral sclerosis (ALS). Methods Thirty-three ALS transgenic mice were used in this study. The brain stem was dissected and collected at the early (95 days), middle (108 days) or late (122 days) stages. The expression of DDX3 and CK1ε in the brain stem of ALS transgenic mice, the distribution and co-localization of positive cells in the hypoglossal nucleus and facial nucleus of the brain stem were detected by RT-PCR, Western blotting and immunofluorescence technology. In each group, the same number of wild type littermates were selected as controls. Results The result of RT-PCR and Western blotting showed that compared with the wild type mice, the expression of DDX3 and CK1ε mRNA in the brain stem of ALS mice remained unchanged at day 95, day 108 and day 122. DDX3 and CK1ε protein levels were up-regulated at day 95 and day 108 but down-regulated at day 122 in the ALS mice brain stem group (P<0.01, P<0.001). The result of immunofluorescence showed that DDX3 and CK1ε positive cells were detected in the sublingual nerve and facial nerve of ALS mice and wild type mice brain stem. DDX3 and CK1ε were expressed in neurons, but not in astrocytes. The immunoreactivity of both DDX3 and CK1ε of ALS mice and wild type mice was different. Conclusion The abnormal expression of DDX3 and CK1ε in the brain stem is closely related to the pathogenesis of ALS.

导出引用

张雅雯王箐原萌刘焕彩王巧真丁昊宇周风华陈燕春. DDX3和酪蛋白激酶1ε在肌萎缩侧索硬化症转基因鼠脑干中的表达变化[J]. 解剖学报. 2018, 49(1): 1-6 https://doi.org/10.16098/j.issn.0529-1356.2018.01.001

ZHANG Ya-wen WANG Qing YUAN Meng LIU Huan-cai WANG Qiao-zhen DING Hao-yu ZHOU Feng-hua CHEN Yan-chun. Expression of DDX3 and casein kinase 1ε in the brain stem of amyotrophic lateral sclerosis transgenic mice[J]. Acta Anatomica Sinica. 2018, 49(1): 1-6 https://doi.org/10.16098/j.issn.0529-1356.2018.01.001

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