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

doi:10.16098/j.issn.0529-1356.2021.05.005

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

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*

1.潍坊医学院生命科学与技术学院生物技术专业； 2.神经疾病与再生修复重点实验室； 3.基础医学院组织学胚胎学教研室，山东潍坊 261053

收稿日期:2020-08-18 修回日期:2020-10-07 出版日期:2021-10-06 发布日期:2021-10-06
通讯作者: 王箐；陈燕春 E-mail:cyc7907@163.com
基金资助:
山东省高校科技发展项目重点项目;山东省高等学校青创科技支持计划;国家级大学生创新训练项目;校级大学生创新基金

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#

1.Biotechnology Specialty, School of Life Science and Technology; 2.Key Laboratory of Neurological Diseases and Regenerative Repair; 3.Department of Histology and Embryology, Basic Medical College, Weifang Medical University, Shandong Weifang 261053,China

Received:2020-08-18 Revised:2020-10-07 Online:2021-10-06 Published:2021-10-06
Contact: WANG Qing；CHEN Yan-chun E-mail:cyc7907@163.com

摘要/Abstract

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。

关键词: 肌萎缩侧索硬化症, DEAD-box解旋酶5, 转录因子12, 海马, 免疫共沉淀技术, SOD1-G93A转基因小鼠

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.

Key words: Amyotrophic lateral sclerosis, mouse DEAD-box helicase 5, Transcription factor 12, Hippocampus, Co-immunoprecipitation assay, SOD1-G93A transgenic mouse

中图分类号:

R329

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.

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.

参考文献

［1］ Crockford C, Newton J, Lonergan K, et al. ALS-specific cognitive and behavior changes associated with advancing disease stage in ALS ［J］. Neurology, 2018, 91(15):e1370-e1380.
［2］ Mathis S, Goizet C, Soulages A, Genetics of amyotrophic lateral sclerosis: a review［J］. J Neurol Sci, 2019, 399:217-226.
［3］ Gurney ME, Pu H, Chiu AY, et al. Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation ［J］.Science,1994,264(5166):1772-1775.
［4］ Mancuso R, Navarro X. Amyotrophic lateral sclerosis: Current perspectives from basic research to the clinic ［J］. Prog Neurobiol, 2015,133:1-26.
［5］ Ballmaier M, Narr KL, Toga AW, et al. Hippocampal morphology and distinguishing late-onset elderly depression［J］. Am J Psychiatry, 2008,165(2):229-237.
［6］ Legrand JMD, Chan AL, La HM, et al. DDX5 plays essential transcriptional and post-transcriptional roles in the maintenance and function of spermatogonia ［J］. Nat Commun, 2019,10(1):2278.
［7］ Hirling H, Scheffner M, Restle T, et al. RNA helicase activity associated with the human p68 protein ［J］. Nature, 1989, 339(6225):562-564.
［8］ Jones K, Wei C, Schoser B, et al. Reduction of toxic RNAs in myotonic dystrophies type 1 and type 2 by the RNA helicase p68/DDX5 ［J］. Proc Natl Acad Sci USA, 2015, 112(26):8041-8045.
［9］ Pu LD，Zhang YW，Wang Q，et al. Expression of DDX3 andcasein kinase1ε in the hippocampus of amyotrophic lateral sclerosistransgenic mice［J］.Acta Anatomica Sinica, 2017, 48(4):375-380. (in Chinese)
蒲蕾东，张雅雯，王箐，等. 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.
［10］ Uittenbogaard M, Chiaramello A. Expression of the bHLH transcription factor Tcf12 (ME1) gene is linked to the expansion of precursor cell populations during neurogenesis ［J］. Brain Res Gene Expr Patterns, 2002, 1(2):115-121.
［11］ Mesman S, Smidt MP. Tcf12 is involved in early cell-fate determination and subset specification of midbrain dopamine neurons ［J］. Front Mol Neurosci, 2017,10:353.
［12］ Wu K, Li S, Bodhinathan K, et al. Enhanced expression of Pctk1, Tcf12 and Ccnd1 in hippocampus of rats: Impact on cognitive function, synaptic plasticity and pathology［J］. Neurobiol Learn Mem, 2012, 97(1):69-80.
［13］ Zhang YW, Wang Q, Yuan M, et al. Expression of DDX3 and casein kinase 1ε in the brain stem ofamyotrophic lateral sclerosis transgenic mice［J］.Acta Anatomica Sinica, 2018, 49(1):1-5. (in Chinese)
张雅雯，王箐，原萌，等. 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):1-5
［14］ Olesen L?, Sivasaravanaparan M, Severino M, et al. Neuron and neuroblast numbers and cytogenesis in the dentate gyrus of aged APP swe /PS1 dE9 transgenic mice: effect of long-term treatment with paroxetine ［J］. Neurobiol Dis, 2017,104:50-60. 
［15］ Takaishi M, Asami T,Yoshida H, et al. Smaller volume of right hippocampal CA2/3 in patients with panic disorder.［J］.Brain Imaging Behav, 2021, 15(1):320-326.
［16］ Lane DP, Hoeffler WK. SV40 large T shares an antigenic determinant with a cellular protein of molecular weight 68,000［J］. Nature, 1980, 288(5787):167-170.
［17］ Giraud G, Terrone S, Bourgeois CF. Functions of DEAD box RNA helicases DDX5 and DDX17 in chromatin organization and transcriptional regulation［J］.BMB Rep, 2018, 51(12):613-622.
［18］ Hoch-Kraft P, White R, Tenzer S, et al. Dual role of the RNA helicase DDX5 in post-transcriptional regulation of myelin basic protein in oligodendrocytes［J］. Cell Sci, 2018, 131(9): jcs204750.
［19］ Nyamao RM, Wu J, Yu L, et al. Roles of DDX5 in the tumorigenesis, proliferation, differentiation, metastasis and pathway regulation of human malignancies［J］. Biochim Biophys Acta Rev Cancer, 2019,1871(1):85-98.
［20］ Labreche K, Simeonova L, Kamoun A, et al. TCF12 is mutated in anaplastic oligodendroglioma ［J］. Nat Commun, 2015, 6:7207.
［21］ Yi S, Yu M, Yang S, et al. Tcf12, A member of basic helix-loop-helix transcription factors, mediates bone marrow mesenchymal stem cell osteogenic differentiation in vitro and in vivo［J］.Stem Cells, 2017, 35(2):386-397.
［22］ Ding HY, Chen YCh, Zheng YW, et al. The expression and localization of transcript factor 12 in cerebral cortex and striatum of SOD1-G93A transgenic mice［J］. Journal of Neuroanatomy, 2019, 35(4): 371-376. (in Chinese)
丁昊宇，陈燕春，郑怡雯，等. 转录因子12在SOD1-G93A转基因小鼠大脑皮层和纹状体中的表达与定位研究［J］.神经解剖学杂志, 2019, 35(4): 371-376
［23］ Zhou F, Zhang C, Guan Y, et al. Screening the expression characteristics of several miRNAs in G93A-SOD1 transgenic mouse: altered expression of miRNA-124 is associated with astrocyte differentiation by targeting Sox2 and Sox9［J］. Neurochem, 2018, 145(1):51-67.

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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

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

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