Optimizing the methodology for precise estimation of skeletal muscle fiber type proportions in humans

Abstract Isolating individual muscle fibers and characterizing their myosin heavy chain (MHC) content using SDS-PAGE has become an increasingly common method for describing skeletal muscle fiber type proportions. In this study, we aimed to assess how the number of muscle fibers analyzed, and whether...

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Main Authors: Nathan Serrano, Matthew R. Buras, Lori R. Roust, Eleanna De Filippis, Christos S. Katsanos
Format: Article
Language:English
Published: Nature Portfolio 2025-08-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-15163-w
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author Nathan Serrano
Matthew R. Buras
Lori R. Roust
Eleanna De Filippis
Christos S. Katsanos
author_facet Nathan Serrano
Matthew R. Buras
Lori R. Roust
Eleanna De Filippis
Christos S. Katsanos
author_sort Nathan Serrano
collection DOAJ
description Abstract Isolating individual muscle fibers and characterizing their myosin heavy chain (MHC) content using SDS-PAGE has become an increasingly common method for describing skeletal muscle fiber type proportions. In this study, we aimed to assess how the number of muscle fibers analyzed, and whether they are characterized in the order of isolation or randomly selected from a larger pool of muscle fibers, affects the precision of fiber type proportion estimates. A total of 170 individual muscle fibers were isolated from vastus lateralis biopsies from each of eight human subjects, and their MHC isoform content was analyzed using SDS-PAGE. To evaluate the precision of fiber type proportion estimates, we employed a resampling approach, varying both the muscle fiber sample size (25, 50, or 100 fibers) and the selection method (ordered vs. random selection). Our results indicate that when analyzing a small number of muscle fibers, precision improves if the fibers are randomly selected from a larger pool rather than characterized in the order they were isolated. These findings have important implications for designing experiments to assess skeletal muscle fiber heterogeneity and its role in health and disease.
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spelling doaj-art-3dd88d8a923e4dafa05d0dbc83bc78a92025-08-20T03:44:06ZengNature PortfolioScientific Reports2045-23222025-08-0115111010.1038/s41598-025-15163-wOptimizing the methodology for precise estimation of skeletal muscle fiber type proportions in humansNathan Serrano0Matthew R. Buras1Lori R. Roust2Eleanna De Filippis3Christos S. Katsanos4School of Life Sciences, Arizona State UniversityDepartment of Quantitative Health Sciences, Mayo Clinic ArizonaCollege of Medicine, Mayo Clinic ArizonaCollege of Medicine, Mayo Clinic ArizonaSchool of Life Sciences, Arizona State UniversityAbstract Isolating individual muscle fibers and characterizing their myosin heavy chain (MHC) content using SDS-PAGE has become an increasingly common method for describing skeletal muscle fiber type proportions. In this study, we aimed to assess how the number of muscle fibers analyzed, and whether they are characterized in the order of isolation or randomly selected from a larger pool of muscle fibers, affects the precision of fiber type proportion estimates. A total of 170 individual muscle fibers were isolated from vastus lateralis biopsies from each of eight human subjects, and their MHC isoform content was analyzed using SDS-PAGE. To evaluate the precision of fiber type proportion estimates, we employed a resampling approach, varying both the muscle fiber sample size (25, 50, or 100 fibers) and the selection method (ordered vs. random selection). Our results indicate that when analyzing a small number of muscle fibers, precision improves if the fibers are randomly selected from a larger pool rather than characterized in the order they were isolated. These findings have important implications for designing experiments to assess skeletal muscle fiber heterogeneity and its role in health and disease.https://doi.org/10.1038/s41598-025-15163-wMuscle fibersMuscle phenotypeMyosin heavy chainAccuracyNumber of fibersSelection of fibers
spellingShingle Nathan Serrano
Matthew R. Buras
Lori R. Roust
Eleanna De Filippis
Christos S. Katsanos
Optimizing the methodology for precise estimation of skeletal muscle fiber type proportions in humans
Scientific Reports
Muscle fibers
Muscle phenotype
Myosin heavy chain
Accuracy
Number of fibers
Selection of fibers
title Optimizing the methodology for precise estimation of skeletal muscle fiber type proportions in humans
title_full Optimizing the methodology for precise estimation of skeletal muscle fiber type proportions in humans
title_fullStr Optimizing the methodology for precise estimation of skeletal muscle fiber type proportions in humans
title_full_unstemmed Optimizing the methodology for precise estimation of skeletal muscle fiber type proportions in humans
title_short Optimizing the methodology for precise estimation of skeletal muscle fiber type proportions in humans
title_sort optimizing the methodology for precise estimation of skeletal muscle fiber type proportions in humans
topic Muscle fibers
Muscle phenotype
Myosin heavy chain
Accuracy
Number of fibers
Selection of fibers
url https://doi.org/10.1038/s41598-025-15163-w
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