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

doi:10.16352/j.issn.1001-6325.2023.02.219

Abstract

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

CLC Number:

R741.02

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.

References 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.

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