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.

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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

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