Methods for assessing mitochondrial quality control mechanisms and cellular consequences in cell culture

Matthew Redmann, Gloria A. Benavides, Willayat Yousuf Wani, Taylor F. Berryhill, Xiaosen Ouyang, Michelle S. Johnson, Saranya Ravi, Kasturi Mitra, Stephen Barnes, Victor M. Darley-Usmar, Jianhua Zhang*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

36 Scopus citations

Abstract

Mitochondrial quality is under surveillance by autophagy, the cell recycling process which degrades and removes damaged mitochondria. Inadequate autophagy results in deterioration in mitochondrial quality, bioenergetic dysfunction, and metabolic stress. Here we describe in an integrated work-flow to assess parameters of mitochondrial morphology, function, mtDNA and protein damage, metabolism and autophagy regulation to provide the framework for a practical assessment of mitochondrial quality. This protocol has been tested with cell cultures, is highly reproducible, and is adaptable to studies when cell numbers are limited, and thus will be of interest to researchers studying diverse physiological and pathological phenomena in which decreased mitochondrial quality is a contributory factor.

Original languageEnglish (US)
Pages (from-to)59-69
Number of pages11
JournalRedox Biology
Volume17
DOIs
StatePublished - Jul 2018

Keywords

  • Autophagy
  • Beta cells
  • Bioenergetics
  • Metabolomics
  • Mitochondrial fragmentation
  • Mitophagy
  • Neuron

ASJC Scopus subject areas

  • Organic Chemistry

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