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Two hours of heat stress induces MAP-kinase signaling and autophagasome accumulation in C2C12 myotubes

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Abstract

Short bouts of heat can induce a hormetic stress response, whereas prolonged or excessive exposure can elicit detrimental effects. We previously demonstrated an increase in autophagic signaling in C2C12 myotubes in response to 1 h of heat at 40 °C. In opposition, longer durations of heat exposure (e.g., 12 and 24 h) lead to an accumulation of autophagasomes and elevations in markers of cellular inflammation, oxidative stress, and apoptosis. Whether a longer, yet moderate, duration of 2 h of heat further enhances autophagic flux and attenuates stress and inflammatory signaling, or transitions the cell toward a dysregulation of autophagy is unclear. In this study, C2C12 myotubes were maintained at 37 °C or exposed to 40 °C (HT) for 2 h, and harvested immediately or following 2, 8, or 24 h of recovery. Two hours of HT immediately increased pAMPK (T172; p = 0.001), and subsequently increased pULK1 (S555) at 2 h of recovery (p = 0.028). LC3 II was increased at 8 h (p = 0.043) and 24 h (p = 0.015) of recovery, whereas p62 was elevated at 2 h (p = 0.002) and 8 h (p < 0.001) of recovery, but returned to baseline by 24 h. In Bafilomycin A1 treated cells, p62 was further increased immediately following HT (p = 0.041). There was also a significant elevation in p-p38 (Thr180/Try182), pJNK (Thr183/Tyr185), and pNFκB (Ser536). These findings suggest that as short as 2 h of heat exposure contributes to cell stress and accumulation of autophagasomes in skeletal muscle.

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All data generated or analyzed during this study are available from the corresponding author on reasonable request.

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Authors and Affiliations

  1. Department of Kinesiology, Iowa State University, Ames, IA, USA

    Corey M. Summers & Rudy J. Valentine

  2. Immunobiology Graduate Program, Iowa State University, Ames, IA, USA

    Corey M. Summers & Rudy J. Valentine

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  1. Corey M. Summers
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  2. Rudy J. Valentine
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Contributions

C.M.S.: Conceived and designed the study, performed experiments, analyzed data, drafted and edited the manuscript. R.J.V.: Conceived and designed the study, performed experiments, analyzed data, edited the manuscript.

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Correspondence to Rudy J. Valentine.

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Summers, C.M., Valentine, R.J. Two hours of heat stress induces MAP-kinase signaling and autophagasome accumulation in C2C12 myotubes. Cell Biochem Biophys 80, 367–373 (2022). https://doi.org/10.1007/s12013-021-01054-0

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