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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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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Author notes

  1. Edited by G. Melino


  1. Department of Physiology & Medical Physics, Royal College of Surgeons in Ireland, Dublin, Ireland

    • Niamh M. C. Connolly
    • , Heiko Düssmann
    • , Orla Watters
    • , Manuela Salvucci
    • , Irene Llorente-Folch
    •  & Jochen H. M. Prehn
  2. Department of Biomedical Sciences, University of Padova, Padova, Italy

    • Pierre Theurey
    • , Paolo Bernardi
    • , Tullio Pozzan
    • , Rosario Rizzuto
    •  & Paola Pizzo
  3. Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary

    • Vera Adam-Vizi
  4. Neuroscience Center of Excellence, School of Medicine, Louisiana State University Health New Orleans, New Orleans, LA, 70112, USA

    • Nicolas G. Bazan
  5. Neuroscience Institute, National Research Council (CNR), Padova, Italy

    • Paolo Bernardi
    • , Tullio Pozzan
    • , Rosario Rizzuto
    •  & Paola Pizzo
  6. University of Salamanca, CIBERFES, Institute of Functional Biology and Genomics (IBFG), CSIC, 37007, Salamanca, Spain

    • Juan P. Bolaños
  7. Institute of Pharmacology and Clinical Pharmacy, University of Marburg, 35043, Marburg, Germany

    • Carsten Culmsee
  8. Neuroregeneration and Stem Cell Programs, Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

    • Valina L. Dawson
  9. Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

    • Valina L. Dawson
  10. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

    • Valina L. Dawson
  11. Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, 21205, USA

    • Valina L. Dawson
  12. Adrienne Helis Malvin Medical Research Foundation, Diana Helis Henry Medical Research Foundation, New Orleans, LA, 70130, USA

    • Valina L. Dawson
  13. Neuroscience Center, University of North Carolina, Chapel Hill, USA

    • Mohanish Deshmukh
  14. Department of Cell and Developmental Biology and Consortium for Mitochondrial Research, University College London, London, WC1E 6BT, UK

    • Michael R. Duchen
  15. Department of Anesthesiology and Center for Shock, Trauma and Anesthesiology Research (STAR), University of Maryland School of Medicine, Baltimore, MD, 21201, USA

    • Gary Fiskum
    •  & Brian M. Polster
  16. Program in Neuroscience, University of Maryland School of Medicine, Baltimore, USA

    • Gary Fiskum
    •  & Brian M. Polster
  17. Unidad de Neuropsicofarmacología Translacional, Complejo Hospitalario Universitario de Albacete, Albacete, Spain

    • Maria F. Galindo
  18. UK Dementia Research Institute at the University of Edinburgh, Edinburgh Medical School, Edinburgh, EH8 9XD, UK

    • Giles E. Hardingham
  19. Department of Molecular Microbiology and Immunology, Johns Hopkins University, Bloomberg School of Public Health, Baltimore, MD, 21205, USA

    • J. Marie Hardwick
  20. Department of Biology, University of Mississippi, University, MS, 38677, USA

    • Mika B. Jekabsons
  21. Department of Internal Medicine, Section of Endocrinology, Yale University, New Haven, CT, USA

    • Elizabeth A. Jonas
  22. Department of Medical Science-Pharmacology, University of Castilla-La Mancha, Albacete, Spain

    • Joaquin Jordán
  23. Neuroscience Translational Center and Depts. of Molecular Medicine and Neuroscience, The Scripps Research Institute, La Jolla, CA, 92037, USA

    • Stuart A. Lipton
  24. Neurodegenerative Disease Center, Scintillon Institute, San Diego, CA, 92121, USA

    • Stuart A. Lipton
  25. Department of Neurosciences, University of California San Diego, School of Medicine, La Jolla, CA, 92093, USA

    • Stuart A. Lipton
  26. Brain and Mind Research Institute, Weill Cornell Medicine, New York, NY, 10065, USA

    • Giovanni Manfredi
  27. National Institute on Aging Intramural Research Program, Baltimore, MD, USA

    • Mark P. Mattson
  28. Department of Neurology, Vanderbilt University School of Medicine, Nashville, TN, USA

    • BethAnn McLaughlin
  29. University Medical Center Mainz, Dept. of Neurology, Mainz, Germany

    • Axel Methner
  30. Department of Pharmacology, University of California, San Diego, CA, 92093, USA

    • Anne N. Murphy
  31. MRC Mitochondrial Biology Unit, University of Cambridge, Cambridge Biomedical Campus, Cambridge, CB2 0XY, UK

    • Michael P. Murphy
  32. Buck Institute for Research on Aging, Novato, CA, 94945, USA

    • David G. Nicholls
  33. Dpto. de Biología Molecular, Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, 28049, Madrid, Spain

    • Jorgina Satrústegui
  34. Brain & Mind Research Institute, University of Ottawa, Ontario, K1H 8M5, Canada

    • Ruth S. Slack
    • , Alvin Joselin
    •  & David S. Park
  35. Department of Neurology, University of California San Francisco and San Francisco Veterans Affairs Medical Center, San Francisco, CA, 94121, USA

    • Raymond A. Swanson
  36. The University of Kansas Alzheimer’s Disease Center and Depts. of Neurology, Molecular and Integrative Physiology, and Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS, 66160, USA

    • Russell H. Swerdlow
  37. Pfizer, Groton, CT, 06340, USA

    • Yvonne Will
  38. Laboratory of Cellular and Molecular Neuropharmacology, Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu, 215021, China

    • Zheng Ying
  39. German Center for Neurodegenerative Diseases (DZNE), Bonn, Germany

    • Anna Gioran
    •  & Daniele Bano
  40. Center for Alzheimer Research, Division of Neurogeriatrics, Department of Neurobiology, Care Sciences and Society, Karolinska Institutet, Stockholm, Sweden

    • Catarina Moreira Pinho
    •  & Maria Ankarcrona


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Conflict of interest

The authors declare that they have no competing interests.

Corresponding author

Correspondence to Jochen H. M. Prehn.

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