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bioRxiv · 10.1101/2025.08.27.670366

Slow- and fast-contracting skeletal muscle fibers have more similar cellular and molecular contractile function at 37°C than at 25°C in older adults

Momb, B. A.·Chipkin, S. R.·Kent, J. A.·Miller, M. S.
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Abstract

As human skeletal muscle cellular and molecular contractile properties are temperature-sensitive, the ability to perform experiments at body temperature ([~]37{degrees}C) may lead to a better understanding of their in vivo responses and potentially their effects upon whole-muscle and whole-body performance. We quantified molecular (myosin-actin cross-bridge mechanics and kinetics) and cellular (specific tension; force divided by cross-sectional area) function in slow-contracting myosin heavy chain (MHC) I and fast-contracting MHC IIA fibers from older adults (n=13, 8 female) at 37{degrees}C and compared these to results at 25{degrees}C. MHC I fibers were more temperature-sensitive than MHC IIA fibers, showing greater increases in cross-bridge kinetics (MHC I: 4.9-8.7x; IIA: 4x) and number or stiffness of strongly-bound cross-bridges (MHC I: 86%; IIA: 34%), leading to increased specific tension in MHC I (19%), with no change in MHC IIA fibers. The expected relationship between fiber force and size (cross-sectional area, CSA) was stronger at 37{degrees}C in both fiber types, explaining 80-82% of the variance compared to 51-52% at 25{degrees}C. Specific tension was unchanged with size at 37{degrees}C in both fiber types, showing that force increases proportionally with CSA, which may be due to the increased number or stiffness of strongly-bound cross-bridges at this temperature. At 25{degrees}C, specific tension decreased with size in agreement with previous experiments. Overall, MHC I and IIA fibers at body temperature (37{degrees}C) became more analogous, including similar specific tension and closer cross-bridge kinetics, and force production was more strongly correlated with fiber size compared to a non-physiological temperature. Abstract Figure O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=103 SRC="FIGDIR/small/670366v1_ufig1.gif" ALT="Figure 1"> View larger version (20K): org.highwire.dtl.DTLVardef@70fe6aorg.highwire.dtl.DTLVardef@4ef5e4org.highwire.dtl.DTLVardef@1f1551forg.highwire.dtl.DTLVardef@10ba6d1_HPS_FORMAT_FIGEXP M_FIG C_FIG KEY POINTS SUMMARYO_LIAlthough skeletal muscle function is highly sensitive to temperature, human single fiber studies have only been conducted at [≤]30{degrees}C. C_LIO_LISmall-amplitude sinusoidal perturbations were utilized to elucidate mechanisms of single fiber force production at human physiological temperature (37{degrees}C). C_LIO_LIWe found that functional differences in slow-contracting myosin heavy chain (MHC I) and fast-contracting MHC IIA fibers observed at 25{degrees}C were less apparent at 37{degrees}C, as force, crossbridge kinetics, and strongly-bound crossbridges increased more in MHC I fibers than MHC IIA fibers at 37 vs. 25{degrees}C. C_LIO_LIThese results indicate that, given the different sensitivity of each fiber type to changes in temperature, functional assessments of muscle should be conducted at 37{degrees}C to better translate to vivo conditions. C_LI

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Momb, B. A., Chipkin, S. R., Kent, J. A., Miller, M. S.. 2025-08-27. Slow- and fast-contracting skeletal muscle fibers have more similar cellular and molecular contractile function at 37°C than at 25°C in older adults. https://doi.org/10.1101/2025.08.27.670366

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