Quantifying Variability and Reliability Associated with High-resolution Respirometry in Human Permeabilized Skeletal Muscle Fibers
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Authors
Ferguson, Emily
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Type
thesis
Language
eng
Keyword
Mitochondria , Oxygraph-2K , Skeletal Muscle
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Abstract
Background: High-resolution respirometry is the primary means by which to measure mitochondrial oxygen consumption in permeabilized human skeletal muscle fibers. In recent years, some research groups have begun examining the degree of variability associated with high-resolution respirometry. However, variability associated with high-resolution respirometry is laboratory specific. Thus, we sought to determine our coefficient of variation (CV), intraclass correlation coefficient (ICC), and technical error of measurement (TEM) values to more accurately adjust our statistical power calculations to account for variability in our Oxygraph-2K measurements for future clinical trials. Purpose: To determine the degree of variability and reliability associated with high-resolution respirometry in human permeabilized skeletal muscle fibers in our laboratory. Hypothesis: We hypothesized that the TEM and CV values for our Oxygraph-2K (O2K) protocol would be less than or similar to those obtained by other research groups conducting exercise interventions in humans. In addition, we hypothesized that there would be no significant differences between respiration values obtained within-trial (intra-biopsy) and between days (inter-biopsy) during our investigation. Methods: Fourteen healthy young recreationally active and trained individuals (age: 19-30 years; female: 10; male: 4; height: 1.73 ± 0.11 m; body mass: 73.0 ± 20.2 kg) participated in this acute skeletal muscle biopsy study. Skeletal muscle biopsies of the vastus lateralis were obtained from the same leg on two experimental visits separated by an average of 6 days. We employed a high-resolution respirometry protocol to examine complex I- and complex II-supported adenosine diphosphate (ADP)-stimulated mitochondrial respiration and creatine sensitivity. Intra- and inter-biopsy TEM, CV, and ICC values were also determined. Results: Intra-biopsy CVs for mass- and mitochondria-specific maximal complex-I and complex-II supported, ADP-stimulated, respiration were 12.7% and 12.6%, respectively. The inter-biopsy CV for mass-specific respiration was 8.1%. The inter-biopsy TEM was 7.4 pmol/sec/mg wet weight and the ICC values were moderate or good for maximal complex I- and II-supported, ADP-stimulated respiration. Respiration supported by various substrates did not differ between visits regardless of creatine condition. Conclusion: Our data are consistent with previously reported CV and TEM values for complex-I and complex-II linked, ADP-stimulated, respiration in human permeabilized skeletal muscle fibers.
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ProQuest PhD and Master's Theses International Dissemination Agreement
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Copying and Preserving Your Thesis
This publication is made available by the authority of the copyright owner solely for the purpose of private study and research and may not be copied or reproduced except as permitted by the copyright laws without written authority from the copyright owner.