Guidelines on experimental methods to assess mitochondrial dysfun.pdf (5.93 MB)

Guidelines on experimental methods to assess mitochondrial dysfunction in cellular models of neurodegenerative diseases.

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posted on 22.11.2019, 16:51 by Niamh M. Connolly, Pierre Theurey, Vera Adam-Vizi, Nicolas G. Bazan, Paolo Bernardi, Juan P. Bolaños, Carsten Culmsee, Valina L. Dawson, Mohanish Deshmukh, Michael R. Duchen, Heiko Düssmann, Gary Fiskum, Maria F. Galindo, Giles E. Hardingham, J Marie Hardwick, Mika B. Jekabsons, Elizabeth A. Jonas, Joaquin Jordán, Stuart A. Lipton, Giovanni Manfredi, Mark P. Mattson, BethAnn McLaughlin, Axel Methner, Anne N. Murphy, Michael P. Murphy, David G. Nicholls, Brian M. Polster, Tullio Pozzan, Rosario Rizzuto, Jorgina Satrústegui, Ruth S. Slack, Raymond A. Swanson, Russell H. Swerdlow, Yvonne Will, Zheng Ying, Alvin Joselin, Anna Gioran, Catarina Moreira Pinho, Orla Watters, Manuela Salvucci, Irene Llorente-Folch, David S. Park, Daniele Bano, Maria Ankarcrona, Paola Pizzo, Jochen HM Prehn

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.

Funding

We acknowledge the support of the CeBioND EU Joint Programme for Neurodegenerative Disease Research (JPND; www.jpnd.eu). The programme is supported through the following national funding organisations: Canada, CIHR; Germany, BMBF; Ireland: Science Foundation Ireland (14/JPND/ B3077); Italy: MIUR; Sweden: VR.

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This article is also available at https://www.nature.com/articles/s41418-017-0020-4

Published Citation

Connolly NMC, Theurey P, Adam-Vizi, V, Bazan NG, Bernardi P, Bolanos JP, et al. Cell Death & Differentiation 2017[Epub ahead of print]

Publication Date

11/12/2017

Publisher

Nature Publishing Group

PubMed ID

29229998

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