DEAD-box RNA解旋酶5和转录因子12与肌萎缩侧索硬化症的关系

林宝勇; 徐进超; 应涵韬; 蒋欣; 刘焕彩; 王箐; 王巧真; 陈燕春

doi:10.16098/j.issn.0529-1356.2021.05.005

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解剖学报 ›› 2021, Vol. 52 ›› Issue (5) : 698-705. DOI: 10.16098/j.issn.0529-1356.2021.05.005

神经生物学

DEAD-box RNA解旋酶5和转录因子12与肌萎缩侧索硬化症的关系

林宝勇^1,2 徐进超^1,2 应涵韬^1,2蒋欣² 刘焕彩² 王箐^2* 王巧真² 陈燕春^2,3*

作者信息 +

Relationship between DEAD-box helicase 5, transcription factor 12 and amyotrophic lateral sclerosis#br#

LIN Bao-yong^1,2 XU Jin-chao^1,2 YING Han-tao^1,2 JIANG Xin² LIU Huan-cai² WANG Qing^2* WANG Qiao-zhen² CHEN Yan-chun^2,3*#br#

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文章历史 +

摘要

目的通过检测DEAD-box RNA解旋酶5（DDX5）和转录因子12（TCF12）在SOD1-G93A突变型肌萎缩侧索硬化症（ALS）转基因小鼠海马中表达情况及其相互作用关系，揭示DDX5和TCF12表达改变与ALS海马病变的关系。方法将42对SOD1-G93A突变型ALS转基因小鼠和野生型小鼠，按照95 d龄（发病早期）、108 d龄（发病中期）和122 d龄（发病晚期）分为3组，通过RT-PCR、Western blotting和免疫荧光双标记技术，检测DDX5和TCF12在海马中的表达情况，通过免疫共沉淀技术检测DDX5和TCF12蛋白之间是否具有相互作用。结果与同龄野生型小鼠相比，在SOD1-G93A突变型ALS转基因小鼠海马中DDX5和TCF12 mRNA无明显变化，而蛋白在 95 d、108 d和122 d表达均上调，差异均有统计学意义。海马齿状回和海马本部均可见DDX5和TCF12阳性细胞，且DDX5和TCF12在海马神经元中表达。SOD1-G93A突变型ALS转基因小鼠海马中DDX5和TCF12免疫阳性反应均较同龄野生型小鼠增强。免疫共沉淀实验检测发现，DDX5 和TCF12蛋白质之间存在相互作用。结论 DDX5和TCF12蛋白在SOD1-G93A突变型ALS转基因小鼠海马组织中表达上调，DDX5和TCF12表达异常与ALS海马组织病变有关。

Abstract

Objective To explore the relationship between the expression of DEAO-box helicase 5(DDX5) and transcription factor 12(TCF12) with amyotrophic lateral sclerosis(ALS) hippocampal lesions by detecting the expressions and the interaction of DDX5 and TCF12 in the hippocampus of SOD1-G93A mutant ALS transgenic mice. Methods Forty-two pairs of SOD1-G93A mutant ALS transgenic mice and wild-type mice were divided into three groups at the age of 95 days (early onset stage), 108 days (middle onset stage) and 122 days (late onset stage). RT-PCR, Western blotting and double immunofluorescence labeled technique were used to detect the expressions of DDX5 and TCF12 in the hippocampus. Co-immunoprecipitation assasy was used to detect the interaction between DDX5 and TCF12. Results Compared with the wild-type mice of the same age, DDX5 and TCF12 mRNA in the hippocampus of SOD1-G93A mutant ALS transgenic mice were unchanged, but DDX5 and TCF12 protein were up-regulated significantly at day 95, 108 and 122. DDX5 and TCF12 positive cells were found in both DG area and hippocampus proper, and DDX5 and TCF12 were co-localized with neurons. The immunoreactivities of DDX5 and TCF12 in the hippocampus of SOD1-G93A mutant transgenic mice were elevated compared with wild-type mice at the same time point. Co-immunoprecipitation assasys confirmed that there existed interactions between DDX5 and TCF12 protein. Conclusion DDX5 and TCF12 protein are up-regulated in the hippocampal tissues of SOD1-G93A mutant ALS transgenic mice. The abnormal expressions of DDX5 and TCF12 are involved in the hippocampal lesions of ALS.

导出引用

林宝勇徐进超应涵韬蒋欣刘焕彩王箐王巧真陈燕春. DEAD-box RNA解旋酶5和转录因子12与肌萎缩侧索硬化症的关系[J]. 解剖学报. 2021, 52(5): 698-705 https://doi.org/10.16098/j.issn.0529-1356.2021.05.005

LIN Bao-yong XU Jin-chao YING Han-tao JIANG Xin LIU Huan-cai WANG Qing WANG Qiao-zhen CHEN Yan-chun. Relationship between DEAD-box helicase 5, transcription factor 12 and amyotrophic lateral sclerosis#br#[J]. Acta Anatomica Sinica. 2021, 52(5): 698-705 https://doi.org/10.16098/j.issn.0529-1356.2021.05.005

中图分类号： R329

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