Quantifying variability associated with high-resolution respirometry in human permeabilized skeletal muscle fibers

Rates of skeletal muscle mitochondrial respiration are often measured using an Oroboros Oxygraph-2K (O2K) high-resolution respirometer. The purpose of this investigation was to determine the coefficients of variation (CV) and technical error of measurement (TEM) for rates of mitochondrial respiratio...

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Main Authors: Emily J. Ferguson, Justin Bureau, Tanner Stokes, Danielle Nyman, Joel Seigel, Shivam Gandhi, Luca J. Delfinis, Brendon J. Gurd, Christopher G.R. Perry, Chris McGlory
Format: Article
Language:English
Published: KeAi Communications Co., Ltd. 2024-09-01
Series:Advanced Exercise and Health Science
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Online Access:http://www.sciencedirect.com/science/article/pii/S2950273X24000298
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author Emily J. Ferguson
Justin Bureau
Tanner Stokes
Danielle Nyman
Joel Seigel
Shivam Gandhi
Luca J. Delfinis
Brendon J. Gurd
Christopher G.R. Perry
Chris McGlory
author_facet Emily J. Ferguson
Justin Bureau
Tanner Stokes
Danielle Nyman
Joel Seigel
Shivam Gandhi
Luca J. Delfinis
Brendon J. Gurd
Christopher G.R. Perry
Chris McGlory
author_sort Emily J. Ferguson
collection DOAJ
description Rates of skeletal muscle mitochondrial respiration are often measured using an Oroboros Oxygraph-2K (O2K) high-resolution respirometer. The purpose of this investigation was to determine the coefficients of variation (CV) and technical error of measurement (TEM) for rates of mitochondrial respiration obtained from two permeabilized skeletal muscle fiber bundles from a single biopsy (intra-biopsy) and on two experimental visits (inter-biopsy) using an O2K high-resolution respirometer. We hypothesized that our CV and TEM values would be similar to those reported by other research groups. We also hypothesized that there would be no statistically significant differences between rates of mitochondrial respiration obtained on two experimental visits separated by one week. Biopsies of the vastus lateralis were obtained from 14 healthy young individuals (female: 10; male: 4; age: 23 ± 3 years; height: 1.73 ± 0.1 m; body mass: 73.3 ± 20.4 kg) from the same leg on two experimental visits separated by an average of 6 ± 2 days. We examined rates of pyruvate and malate-supported, adenosine diphosphate (ADP)-stimulated mitochondrial respiration in the presence and absence of 20 mM creatine. Mean average intra- and inter-biopsy CVs for maximal complex I+II-supported, ADP-stimulated mitochondrial respiration were 11.5 % and 9.7 %, respectively. Mean average intra- and inter-biopsy TEM values for maximal complex I+II-supported, ADP-stimulated mitochondrial respiration were 10.6 pmol/s/mg wet weight and 8.6 pmol/s/mg wet weight, respectively. Statistically significant differences for rates of mitochondrial respiration were not observed between experimental visits regardless of creatine condition (non-creatine or 20 mM creatine). Our data are consistent with previously reported CV and TEM values and demonstrate that changes in maximal mitochondrial respiration of approximately 15 % can be reliably detected with a sample size of ∼10 participants.
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spelling doaj-art-0d9c0a139e3f441186cae83bbb095d312025-08-20T03:21:12ZengKeAi Communications Co., Ltd.Advanced Exercise and Health Science2950-273X2024-09-011317017710.1016/j.aehs.2024.05.006Quantifying variability associated with high-resolution respirometry in human permeabilized skeletal muscle fibersEmily J. Ferguson0Justin Bureau1Tanner Stokes2Danielle Nyman3Joel Seigel4Shivam Gandhi5Luca J. Delfinis6Brendon J. Gurd7Christopher G.R. Perry8Chris McGlory9School of Kinesiology and Health Studies, Queen’s University, Kingston, Ontario, CanadaSchool of Kinesiology and Health Studies, Queen’s University, Kingston, Ontario, CanadaSchool of Kinesiology and Health Studies, Queen’s University, Kingston, Ontario, CanadaSchool of Kinesiology and Health Studies, Queen’s University, Kingston, Ontario, CanadaSchool of Kinesiology and Health Studies, Queen’s University, Kingston, Ontario, CanadaMuscle Health Research Centre, School of Kinesiology and Health Science, York University, Toronto, Ontario, CanadaMuscle Health Research Centre, School of Kinesiology and Health Science, York University, Toronto, Ontario, CanadaSchool of Kinesiology and Health Studies, Queen’s University, Kingston, Ontario, CanadaMuscle Health Research Centre, School of Kinesiology and Health Science, York University, Toronto, Ontario, CanadaSchool of Kinesiology and Health Studies, Queen’s University, Kingston, Ontario, Canada; Department of Medicine, Queen’s University, Kingston, Ontario, Canada; Correspondence to: School of Kinesiology and Health Studies, Queen’s University, 28 Division Street, Kingston, Ontario, Canada.Rates of skeletal muscle mitochondrial respiration are often measured using an Oroboros Oxygraph-2K (O2K) high-resolution respirometer. The purpose of this investigation was to determine the coefficients of variation (CV) and technical error of measurement (TEM) for rates of mitochondrial respiration obtained from two permeabilized skeletal muscle fiber bundles from a single biopsy (intra-biopsy) and on two experimental visits (inter-biopsy) using an O2K high-resolution respirometer. We hypothesized that our CV and TEM values would be similar to those reported by other research groups. We also hypothesized that there would be no statistically significant differences between rates of mitochondrial respiration obtained on two experimental visits separated by one week. Biopsies of the vastus lateralis were obtained from 14 healthy young individuals (female: 10; male: 4; age: 23 ± 3 years; height: 1.73 ± 0.1 m; body mass: 73.3 ± 20.4 kg) from the same leg on two experimental visits separated by an average of 6 ± 2 days. We examined rates of pyruvate and malate-supported, adenosine diphosphate (ADP)-stimulated mitochondrial respiration in the presence and absence of 20 mM creatine. Mean average intra- and inter-biopsy CVs for maximal complex I+II-supported, ADP-stimulated mitochondrial respiration were 11.5 % and 9.7 %, respectively. Mean average intra- and inter-biopsy TEM values for maximal complex I+II-supported, ADP-stimulated mitochondrial respiration were 10.6 pmol/s/mg wet weight and 8.6 pmol/s/mg wet weight, respectively. Statistically significant differences for rates of mitochondrial respiration were not observed between experimental visits regardless of creatine condition (non-creatine or 20 mM creatine). Our data are consistent with previously reported CV and TEM values and demonstrate that changes in maximal mitochondrial respiration of approximately 15 % can be reliably detected with a sample size of ∼10 participants.http://www.sciencedirect.com/science/article/pii/S2950273X24000298Mitochondrial respirationHuman skeletal muscleVariability
spellingShingle Emily J. Ferguson
Justin Bureau
Tanner Stokes
Danielle Nyman
Joel Seigel
Shivam Gandhi
Luca J. Delfinis
Brendon J. Gurd
Christopher G.R. Perry
Chris McGlory
Quantifying variability associated with high-resolution respirometry in human permeabilized skeletal muscle fibers
Advanced Exercise and Health Science
Mitochondrial respiration
Human skeletal muscle
Variability
title Quantifying variability associated with high-resolution respirometry in human permeabilized skeletal muscle fibers
title_full Quantifying variability associated with high-resolution respirometry in human permeabilized skeletal muscle fibers
title_fullStr Quantifying variability associated with high-resolution respirometry in human permeabilized skeletal muscle fibers
title_full_unstemmed Quantifying variability associated with high-resolution respirometry in human permeabilized skeletal muscle fibers
title_short Quantifying variability associated with high-resolution respirometry in human permeabilized skeletal muscle fibers
title_sort quantifying variability associated with high resolution respirometry in human permeabilized skeletal muscle fibers
topic Mitochondrial respiration
Human skeletal muscle
Variability
url http://www.sciencedirect.com/science/article/pii/S2950273X24000298
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