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

蒲蕾东 张雅雯 王箐 刘焕彩 刘永新 王巧真 管英俊 陈燕春

解剖学报 ›› 2017, Vol. 48 ›› Issue (4) : 375-380.

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解剖学报 ›› 2017, Vol. 48 ›› Issue (4) : 375-380. DOI: 10.16098/j.issn.0529-1356.2017.04.002
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海马中的表达

  • 蒲蕾东1 张雅雯1 王箐2 刘焕彩3 刘永新1 王巧真2 管英俊1 陈燕春1*
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Expression of DDX3 and casein kinase 1ε in the hippocampus of the amyotrophic lateral sclerosis transgenic mice

  • PU Lei-dong1 ZHANG Ya-wen1 WANG Qing2 LIU Huan-cai3 LIU Yong-xin1 WANG Qiao-zhen2 GUAN Ying-jun1 CHEN Yan-chun1*
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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;CK1ε mRNA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CK1ε mRNA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 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 CK1ε is closely related to hippocampus lesion in ALS.

关键词

肌萎缩侧索硬化症 / 海马 / DDX3 / 酪蛋白激酶1ε / 转基因 / 免疫印迹法 / 小鼠

Key words

Amyotrophic lateral sclerosis / Hippocampus / DDX3 / Casein kinase 1ε / Transgenic / Western blotting / Mouse

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蒲蕾东 张雅雯 王箐 刘焕彩 刘永新 王巧真 管英俊 陈燕春. 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[J]. 解剖学报. 2017, 48(4): 375-380 https://doi.org/10.16098/j.issn.0529-1356.2017.04.002
PU Lei-dong ZHANG Ya-wen WANG Qing LIU Huan-cai LIU Yong-xin WANG Qiao-zhen GUAN Ying-jun CHEN Yan-chun. Expression of DDX3 and casein kinase 1ε in the hippocampus of the amyotrophic lateral sclerosis transgenic mice[J]. Acta Anatomica Sinica. 2017, 48(4): 375-380 https://doi.org/10.16098/j.issn.0529-1356.2017.04.002

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基金

国家自然科学基金;国家自然科学基金;山东省自然科学基金;山东省自然科学基金;山东省科技发展计划项目;山东省教育厅课题;山东省教育厅课题

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