α-突触核蛋白/A53T突变体损伤SH-SY5Y细胞的MAM并导致线粒体结构异常及自噬

doi:10.16352/j.issn.1001-6325.2023.02.219

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (2): 219-224.doi: 10.16352/j.issn.1001-6325.2023.02.219

α-突触核蛋白/A53T突变体损伤SH-SY5Y细胞的MAM并导致线粒体结构异常及自噬

岳静静, 伍超, 高歌, 鲁玲玲^*

首都医科大学基础医学院神经生物学系, 北京 100069

收稿日期:2021-12-06 修回日期:2022-07-14 出版日期:2023-02-05 发布日期:2023-02-02
通讯作者: ^*lllu@ccmu.edu.cn
基金资助:
国家自然科学基金重点项目(81830033);北京市自然科学基金(7192017)

Alpha-synuclein /A53T mutant damages the mitochondria associated membrane and results in mitochondrial structural abnormalities and autophagy of neuroblastoma cell line SH-SY5Y

YUE Jingjing, WU Chao, GAO Ge, LU Lingling^*

Department of Neurobiology, Capital Medical University, Beijing 100069, China

Received:2021-12-06 Revised:2022-07-14 Online:2023-02-05 Published:2023-02-02
Contact: ^*lllu@ccmu.edu.cn

摘要/Abstract

摘要： 目的研究α-突触核蛋白(α-syn)A53T突变体对人神经母细胞瘤细胞内质网线粒体相关膜(MAM)的影响,进而探讨α-syn突变后导致神经元毒性的分子细胞机制。方法首先构建过表达α-syn/A53T突变基因的人神经母细胞瘤细胞系(SH-SY5Y)细胞,然后利用分子邻位杂交(PLA)的方法标记MAM并计数MAM的数量。用Rhod2标记线粒体钙离子,动态观测线粒体钙离子变化情况。用激光共聚焦显微镜及电镜技术进一步观察线粒体的结构。结果过表达α-syn/A53T突变体可以导致明显的细胞毒性(P＜0.001),MAM的数量显著减少(P＜0.001),MAM钙离子转运受到抑制。线粒体变为短棒状、球状,并在神经细胞内出现线粒体空泡以及自噬体。结论 α-syn/A53T突变体可以通过破坏SH-SY5Y细胞的MAM导致线粒体结构异常并自噬。

关键词: α-突触核蛋白, 内质网线粒体相关膜, 线粒体, 帕金森病

Abstract: Objective To investigate the effect of α-synuclein (α-syn) A53T mutant on mitochondria associated membrane (MAM) of endoplasmic reticulum, and to explore the molecular and cellular mechanism of neuronal toxicity induced by α-syn mutation. Methods Human neuroblastoma cell line SH-SY5Y with over expressed α-syn/A53T mutation gene, was constructed. MAM was labeled by proximity ligation assay and the number of MAM was counted. Mitochondrial calcium ions were labeled with Rhod2 and the changes of mitochondrial calcium ions were dynamically observed. The structure of mitochondria was further observed by laser confocal microscopy and electron microscopy. Results Over-expression of α-syn/A53T mutants resulted in significant cytotoxicity, inhibition of MAM calcium transport, and short rod-shaped and globular mitochondria as found by confocal laser microscopy. Mitochondrial vacuoles and autophagy were observed by electron microscopy. Conclusions α -syn/A53T mutants can cause mitochondrial structural abnormalities and autophagy by destroying MAM.

Key words: α-synuclein, mitochondria associated membrane of endoplasmic reticulum, mitochondria, Parkinson's disease

中图分类号:

R741.02

岳静静, 伍超, 高歌, 鲁玲玲. α-突触核蛋白/A53T突变体损伤SH-SY5Y细胞的MAM并导致线粒体结构异常及自噬[J]. 基础医学与临床, 2023, 43(2): 219-224.

YUE Jingjing, WU Chao, GAO Ge, LU Lingling. Alpha-synuclein /A53T mutant damages the mitochondria associated membrane and results in mitochondrial structural abnormalities and autophagy of neuroblastoma cell line SH-SY5Y[J]. Basic & Clinical Medicine, 2023, 43(2): 219-224.

参考文献 14

[1]	郭丽丽, 王咏, 王莹, 等. SNCA和GBA在帕金森病中相互作用研究进展[J]. 基础医学与临床, 2019, 39: 97-101.
[2]	Riederer P, Berg D, Casadei N, et al. α-Synuclein in Parkinson's disease: causal or bystander?[J]. J Neural Transm (Vienna), 2019, 126: 815-840.
[3]	Lee MK, Stirling W, Xu Y, et al. Human alpha-synuclein-harboring familial Parkinson's disease-linked A53T mutation causes neurodegenerative disease with alpha-synuclein aggregation in transgenic mice[J]. Proc Natl Acad Sci U S A, 2002, 99: 8968-8973.
[4]	Portz P, Lee MK. Changes in drp1 function and mitochondrial morphology are associated with the α-Synuclein pathology in a transgenic mouse model of Parkinson's disease[J]. Cells, 2021, 10, 885-892.
[5]	Paillusson S, Gomez-Suaga P, Stoica R, et al. α-Synuclein binds to the ER-mitochondria tethering protein VAPB to disrupt Ca2+ homeostasis and mitochondrial ATP production[J]. Acta Neuropathol, 2017, 134: 129-149.
[6]	Lu L, Zhang C, Cai Q, et al. Voltage-dependent anion channel involved in the α-synuclein-induced dopaminergic neuron toxicity in rats[J]. Acta Biochim Biophys Sin (Shanghai), 2013, 45: 170-178.
[7]	Calì T, Ottolini D, Vicario M, et al. SplitGFP technology reveals dose-dependent ER-mitochondria interface modulation by α-Synuclein A53T and A30P mutants[J]. Cells, 2019, 8, 1072-1090.
[8]	Giasson BI, Duda JE, Quinn SM, et al. Neuronal alpha-synucleinopathy with severe movement disorder in mice expressing A53T human alpha-synuclein[J]. Neuron, 2002, 34: 521-533.
[9]	Janikiewicz J, Szymański J, Malinska D, et al. Mitochondria-associated membranes in aging and senescence: structure, function, and dynamics[J]. Cell Death Dis, 2018, 9: 332-344.
[10]	Han H, Tan J, Wang R, et al. PINK1 phosphorylates Drp1(S616) to regulate mitophagy-independent mitochondrial dynamics[J]. EMBO Rep, 2020, 21: 48686-48693.
[11]	Maki RA, Holzer M, Motamedchaboki K, et al. Human myeloperoxidase (hMPO) is expressed in neurons in the substantia nigra in Parkinson's disease and in the hMPO-α-synuclein-A53T mouse model, correlating with increased nitration and aggregation of α-synuclein and exacerbation of motor impairment[J]. Free Radic Biol Med, 2019, 141: 115-140.
[12]	Hu D, Sun X, Liao X, et al. Alpha-synuclein suppresses mitochondrial protease ClpP to trigger mitochondrial oxidative damage and neurotoxicity[J]. Acta Neuropathol, 2019, 137: 939-960.
[13]	Szabadkai G, Bianchi K, Várnai P, et al. Chaperone-mediated coupling of endoplasmic reticulum and mitochondrial Ca2+ channels[J]. J Cell Biol, 2006, 175: 901-911.
[14]	Yoo SM, Jung YK. A molecular approach to mitophagy and mitochondrial dynamics[J]. Mol Cell, 2018, 41: 18-26.

[1]	何綦琪, 吴晃, 李昱卓, 包军胜. 线粒体功能障碍在肾结石成石中作用的研究进展[J]. 基础医学与临床, 2023, 43(2): 306-310.
[2]	刘立洋, 柳青, 刘雅萍, 张学. 线粒体功能障碍在肌萎缩侧索硬化发病中作用的研究进展[J]. 基础医学与临床, 2023, 43(2): 332-335.
[3]	左赋兴, 苑青, 蔡洪庆, 韩睿钦, 万经海. 新型微量注射装置在建立帕金森病模型小鼠中的应用[J]. 基础医学与临床, 2022, 42(7): 1031-1034.
[4]	连婷, 张瑞君, 熊晓兰, 张小雅, 余涛, 张世忠. 线粒体通透性转换孔在锌缺乏模型大鼠心肌缺血/再灌注损伤中的作用[J]. 基础医学与临床, 2022, 42(3): 389-394.
[5]	张琦悦, 张伟丽. 血管老化在动脉粥样硬化中的研究进展[J]. 基础医学与临床, 2022, 42(3): 372-377.
[6]	吴长宝, 郝延磊. STAT3与帕金森病发病机制的研究进展[J]. 基础医学与临床, 2022, 42(12): 1935-1938.
[7]	祁俊清, 曹梦菲, 赵倩茹, 尹俊. 线粒体SIRT3在器官纤维化中作用的研究进展[J]. 基础医学与临床, 2022, 42(11): 1799-1803.
[8]	江罗佳, 许海波. 白藜芦醇缓解低氧/复氧诱导的TCMK1细胞线粒体自噬[J]. 基础医学与临床, 2022, 42(11): 1709-1715.
[9]	黄雨晴, 高洪婷, 杨黎星, 陈秋霞, 王钰铖, 鞠瑞, 郭磊. 羧胺三唑乳清酸盐抑制小鼠胶质瘤细胞系GL261增殖及线粒体能量代谢[J]. 基础医学与临床, 2021, 41(7): 957-962.
[10]	马家玲, 高艳平. miR-183靶向Bnip3l对缺氧/复氧的小鼠海马神经元细胞线粒体自噬的影响[J]. 基础医学与临床, 2021, 41(5): 698-703.
[11]	张欣, 廖晓辉. 线粒体功能障碍在急性肾损伤向慢性肾脏疾病转变中的作用[J]. 基础医学与临床, 2021, 41(4): 589-592.
[12]	李梦婕, 柯本, 龙脉, 房向东. 线粒体融合蛋白Mfn2在糖尿病肾病中作用的研究进展[J]. 基础医学与临床, 2021, 41(12): 1823-1827.
[13]	胡惟恺, 刘瑞霞, 阴赪宏. 人端粒酶反转录酶线粒体转位的研究进展[J]. 基础医学与临床, 2021, 41(12): 1818-1822.
[14]	王一川, 刘小静, 叶峰, 杨雪亮, 陈娜, 吴剑华, 李彦霖. 线粒体移植的研究进展[J]. 基础医学与临床, 2021, 41(12): 1838-1842.
[15]	马利芳, 梁建庆, 何建成, 孙雪, 梁鹏. 谷氨酸转运体在帕金森病发病机制中作用的研究进展[J]. 基础医学与临床, 2021, 41(12): 1813-1817.

α-突触核蛋白/A53T突变体损伤SH-SY5Y细胞的MAM并导致线粒体结构异常及自噬

Alpha-synuclein /A53T mutant damages the mitochondria associated membrane and results in mitochondrial structural abnormalities and autophagy of neuroblastoma cell line SH-SY5Y

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献 14

相关文章 15

编辑推荐

Metrics

本文评价