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Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements
Skeletal muscle mitochondria play a specific role in many disease pathologies. As such, the measurement of oxygen consumption as an indicator of mitochondrial function in this tissue has become more prevalent. Although many technologies and assays exist that measure mitochondrial respiratory pathway...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MyJove Corporation
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4692683/ https://www.ncbi.nlm.nih.gov/pubmed/26555567 http://dx.doi.org/10.3791/53216 |
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author | Boutagy, Nabil E. Rogers, George W. Pyne, Emily S. Ali, Mostafa M. Hulver, Matthew W. Frisard, Madlyn I. |
author_facet | Boutagy, Nabil E. Rogers, George W. Pyne, Emily S. Ali, Mostafa M. Hulver, Matthew W. Frisard, Madlyn I. |
author_sort | Boutagy, Nabil E. |
collection | PubMed |
description | Skeletal muscle mitochondria play a specific role in many disease pathologies. As such, the measurement of oxygen consumption as an indicator of mitochondrial function in this tissue has become more prevalent. Although many technologies and assays exist that measure mitochondrial respiratory pathways in a variety of cells, tissue and species, there is currently a void in the literature in regards to the compilation of these assays using isolated mitochondria from mouse skeletal muscle for use in microplate based technologies. Importantly, the use of microplate based respirometric assays is growing among mitochondrial biologists as it allows for high throughput measurements using minimal quantities of isolated mitochondria. Therefore, a collection of microplate based respirometric assays were developed that are able to assess mechanistic changes/adaptations in oxygen consumption in a commonly used animal model. The methods presented herein provide step-by-step instructions to perform these assays with an optimal amount of mitochondrial protein and reagents, and high precision as evidenced by the minimal variance across the dynamic range of each assay. |
format | Online Article Text |
id | pubmed-4692683 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | MyJove Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-46926832016-01-07 Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements Boutagy, Nabil E. Rogers, George W. Pyne, Emily S. Ali, Mostafa M. Hulver, Matthew W. Frisard, Madlyn I. J Vis Exp Developmental Biology Skeletal muscle mitochondria play a specific role in many disease pathologies. As such, the measurement of oxygen consumption as an indicator of mitochondrial function in this tissue has become more prevalent. Although many technologies and assays exist that measure mitochondrial respiratory pathways in a variety of cells, tissue and species, there is currently a void in the literature in regards to the compilation of these assays using isolated mitochondria from mouse skeletal muscle for use in microplate based technologies. Importantly, the use of microplate based respirometric assays is growing among mitochondrial biologists as it allows for high throughput measurements using minimal quantities of isolated mitochondria. Therefore, a collection of microplate based respirometric assays were developed that are able to assess mechanistic changes/adaptations in oxygen consumption in a commonly used animal model. The methods presented herein provide step-by-step instructions to perform these assays with an optimal amount of mitochondrial protein and reagents, and high precision as evidenced by the minimal variance across the dynamic range of each assay. MyJove Corporation 2015-10-30 /pmc/articles/PMC4692683/ /pubmed/26555567 http://dx.doi.org/10.3791/53216 Text en Copyright © 2015, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/ |
spellingShingle | Developmental Biology Boutagy, Nabil E. Rogers, George W. Pyne, Emily S. Ali, Mostafa M. Hulver, Matthew W. Frisard, Madlyn I. Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements |
title | Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements |
title_full | Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements |
title_fullStr | Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements |
title_full_unstemmed | Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements |
title_short | Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements |
title_sort | using isolated mitochondria from minimal quantities of mouse skeletal muscle for high throughput microplate respiratory measurements |
topic | Developmental Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4692683/ https://www.ncbi.nlm.nih.gov/pubmed/26555567 http://dx.doi.org/10.3791/53216 |
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