Guidelines on experimental methods to assess mitochondrial dysfunction in cellular models of neurodegenerative diseases.
Guidelines on experimental methods to assess mitochondrial dysfunction in cellular models of neurodegenerative diseases.
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DOI：
10.1038/s41418-017-0020-4
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发表时间：
2018-03
影响因子：
12.4
通讯作者：
Prehn JHM
中科院分区：
文献类型：
--
作者：
Connolly NMC;Theurey P;Adam-Vizi V;Bazan NG;Bernardi P;Bolaños JP;Culmsee C;Dawson VL;Deshmukh M;Duchen MR;Düssmann H;Fiskum G;Galindo MF;Hardingham GE;Hardwick JM;Jekabsons MB;Jonas EA;Jordán J;Lipton SA;Manfredi G;Mattson MP;McLaughlin B;Methner A;Murphy AN;Murphy MP;Nicholls DG;Polster BM;Pozzan T;Rizzuto R;Satrústegui J;Slack RS;Swanson RA;Swerdlow RH;Will Y;Ying Z;Joselin A;Gioran A;Moreira Pinho C;Watters O;Salvucci M;Llorente-Folch I;Park DS;Bano D;Ankarcrona M;Pizzo P;Prehn JHM
神经退行性疾病是一种以神经元进行性退化和死亡为特征的慢性衰弱疾病。线粒体功能障碍与大多数神经退行性疾病有关，但在许多情况下，这种功能障碍是潜在病理的原因还是结果，以及它是否代表一个可行的治疗靶点，目前尚不清楚。因此，迫切需要利用和优化适当的细胞模型和实验技术来确定线粒体功能障碍对神经退行性疾病表型的贡献。在这篇共识文章中，我们整理并讨论了在神经退行性疾病的体外细胞模型中评估线粒体功能的现有实验方法的细节和缺陷，包括测量原代神经元培养中的氧耗率的特定协议，以及线粒体膜电位和线粒体NAD(P)H的单神经元、延时荧光成像。作为神经退行性疾病细胞生物能量学(CeBioND)联盟(www.cebiond.org)的一部分，我们正在进行跨疾病分析，以确定在阿尔茨海默病、帕金森病和亨廷顿病的细胞模型中线粒体生物能量障碍所涉及的共同和不同的分子机制。在这里，我们提供了CeBioND联盟的五个合作实验室的标准化的详细指南和协议，以及该领域其他专家的额外贡献。
Neurodegenerative diseases are a spectrum of chronic, debilitating disorders characterised by the progressive degeneration and death of neurons. Mitochondrial dysfunction has been implicated in most neurodegenerative diseases, but in many instances it is unclear whether such dysfunction is a cause or an effect of the underlying pathology, and whether it represents a viable therapeutic target. It is therefore imperative to utilise and optimise cellular models and experimental techniques appropriate to determine the contribution of mitochondrial dysfunction to neurodegenerative disease phenotypes. In this consensus article, we collate details on and discuss pitfalls of existing experimental approaches to assess mitochondrial function in in vitro cellular models of neurodegenerative diseases, including specific protocols for the measurement of oxygen consumption rate in primary neuron cultures, and single-neuron, time-lapse fluorescence imaging of the mitochondrial membrane potential and mitochondrial NAD(P)H. As part of the Cellular Bioenergetics of Neurodegenerative Diseases (CeBioND) consortium (www.cebiond.org), we are performing cross-disease analyses to identify common and distinct molecular mechanisms involved in mitochondrial bioenergetic dysfunction in cellular models of Alzheimer’s, Parkinson’s, and Huntington’s diseases. Here we provide detailed guidelines and protocols as standardised across the five collaborating laboratories of the CeBioND consortium, with additional contributions from other experts in the field.
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影响因子：
3
作者：
Bilan, Dmitry S.;Matlashov, Mikhail E.;Gorokhovatsky, Andrey Yu.;Schultz, Carsten;Enikolopov, Grigori;Belousov, Vsevolod V.
通讯作者：
Belousov, Vsevolod V.
影响因子：
82.9
作者：
Breckwoldt, Michael;Pfister, Franz M. J.;Misgeld, Thomas
通讯作者：
Misgeld, Thomas
影响因子：
4.2
作者：
Aouacheria A;Baghdiguian S;Lamb HM;Huska JD;Pineda FJ;Hardwick JM
通讯作者：
Hardwick JM
影响因子：
7.4
作者：
Blacker, Thomas S.;Duchen, Michael R.
通讯作者：
Duchen, Michael R.
影响因子：
3.7
作者：
Biffi E;Regalia G;Menegon A;Ferrigno G;Pedrocchi A
通讯作者：
Pedrocchi A