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

doi:10.1016/j.redox.2018.04.005

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

Neurology

Research output: Contribution to journal › Article › peer-review

43 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 language	English (US)
Pages (from-to)	59-69
Number of pages	11
Journal	Redox Biology
Volume	17
DOIs	https://doi.org/10.1016/j.redox.2018.04.005
State	Published - Jul 2018

Funding

This work was partially supported by UAB Health Service Foundation General Endowment Fund , UAB-UCSD O′Brien Acute Kidney Injury Center ( P30 DK079337 ), UAB Blue Sky program, UAB AMC21 reload multi-investigator grant, Nathan Shock Center P30 G050886 , and NIHR01-NS064090 . The mass spectrometer was purchased with a grant from the UAB Health Services Foundation General Endownemt Fund.

Keywords

Autophagy
Beta cells
Bioenergetics
Metabolomics
Mitochondrial fragmentation
Mitophagy
Neuron

ASJC Scopus subject areas

Organic Chemistry

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.redox.2018.04.005

Cite this

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title = "Methods for assessing mitochondrial quality control mechanisms and cellular consequences in cell culture",

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",

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doi = "10.1016/j.redox.2018.04.005",

language = "English (US)",

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AU - Redmann, Matthew

AU - Benavides, Gloria A.

AU - Wani, Willayat Yousuf

AU - Berryhill, Taylor F.

AU - Ouyang, Xiaosen

AU - Johnson, Michelle S.

AU - Ravi, Saranya

AU - Mitra, Kasturi

AU - Barnes, Stephen

AU - Darley-Usmar, Victor M.

AU - Zhang, Jianhua

PY - 2018/7

Y1 - 2018/7

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AB - 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.

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KW - Beta cells

KW - Bioenergetics

KW - Metabolomics

KW - Mitochondrial fragmentation

KW - Mitophagy

KW - Neuron

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

Abstract

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

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