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Redox Biol. 2018 Jul;17:59-69. doi: 10.1016/j.redox.2018.04.005. Epub 2018 Apr 10.

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

Author information

1
Department of Pathology and Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, AL 35294, United States.
2
Department of Pharmacology and Toxicology and Targeted Metabolomics & Proteomics Laboratory, University of Alabama at Birmingham, Birmingham, AL 35294, United States.
3
Department of Genetics, University of Alabama at Birmingham, Birmingham, AL 35294, United States.
4
Department of Pathology and Center for Free Radical Biology, University of Alabama at Birmingham, Birmingham, AL 35294, United States; VA Medical Center, University of Alabama at Birmingham, Birmingham, AL 35294, United States. Electronic address: jianhuazhang@uabmc.edu.

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.

KEYWORDS:

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

PMID:
29677567
PMCID:
PMC6006680
DOI:
10.1016/j.redox.2018.04.005
[Indexed for MEDLINE]
Free PMC Article

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