Cell Stress and Chaperones

, Volume 23, Issue 5, pp 871–883 | Cite as

Heat acclimation increases mitochondrial respiration capacity of C2C12 myotubes and protects against LPS-mediated energy deficit

  • Meghan G. Patton
  • Trevor L. Gillum
  • Mandy C. Szymanski
  • Lacey M. Gould
  • Claire J. Lauterbach
  • Roger A. Vaughan
  • Matthew R. Kuennen
Original Paper


This work investigated the effect of a 6-day heat acclimation (HA) protocol on myotube metabolic responses at baseline and in response to a subsequent lipopolysaccharide (LPS) challenge. C2C12 myotubes were incubated for 2 h/day at 40 °C for 6 days (HA) or maintained at 37 °C (C). Following 24-h recovery, myotubes were challenged with 500 ng/ml LPS for 2 h, then collected for analysis of protein markers of mitochondrial biogenesis and macronutrient storage. Functional significance of these changes was confirmed with mitochondrial respiration and glycolytic measurements on a Seahorse XF-96 analyzer. HA stimulated mitochondrial biogenesis and increased indicators of mitochondrial content [SIRT1 (+ 62%); PGC-1α (+ 57%); NRF-1 (+ 40%); TFAM (+ 141%); CS (+ 25%); CytC (+ 38%); all p < 0.05]. Altered lipid biosynthesis enzymes [p-ACCa:ACC (+ 59%; p = 0.04) and FAS (− 86%; p < 0.01)] suggest fatty acid generation may have been downregulated, whereas increased GLUT4 (+ 69%; p < 0.01) and LDH-B (+ 366%; p < 0.01) suggest aerobic glycolytic capacity may have been improved. Mitochondrial biogenesis signaling in HA myotubes was suppressed by 500 ng/ml LPS (PGC-1α, NRF-1, TFAM; all p > 0.05) but increased LDH-B (+ 30%; p = 0.02) and CPT-1 (+ 55%; p < 0.01) suggesting improved catabolic function. Basal respiration was increased in HA myotubes (+ 8%; p < 0.01) and HA myotubes maintained elevated basal respiration during LPS challenge (+ 8%; p < 0.01). LPS reduced peak respiration in C myotubes (− 6%; p < 0.01) but did not impair peak respiration in HA myotubes (p > 0.05). Oxidative reliance was elevated in HA over that in control (+ 25%; p < 0.01) and in HA + LPS over C + LPS (+ 30%; p < 0.01). In summary, HA stimulated mitochondrial biogenesis in C2C12 myotubes. HA myotubes exhibited (1) elevated basal/peak mitochondrial respiration capacities; (2) greater oxidative reliance; and (3) protection against LPS-mediated respiration impairment. Collectively, these data suggest HA may improve aerobic metabolism in skeletal muscle and protect against LPS-mediated energy deficit.


Hyperthermia Lipopolysaccharide Oxidative metabolism Oxidative reliance 



Phosphorylated acetyl-CoA carboxylase


Acetyl-CoA carboxylase


Phosphorylated 5′-AMP-activated protein kinase


5′-AMP-activated protein kinase


Carnitine palmitoyltransferase I


Citrate synthase


Cytochrome c


Fatty acid synthase


Forkhead in rhabdomyosarcoma


Glucose uptake transporter 4


Glycogen synthase


Phosphorylated glycogen synthase kinase 3 alpha


Phosphorylated glycogen synthase kinase 3 beta


Glycogen synthase kinase 3 alpha/beta


Lactate dehydrogenase A


Lactate dehydrogenase B


Myosin heavy chain IIB


Myocyte differentiation factor


Mitochondrial transcription factor A


Troponin C


Nuclear respiratory factor 1


Peroxisome proliferator-associated receptor gamma co-activator-1

PPAR β/δ

Peroxisome proliferator-associated receptor beta/delta


Peroxisome proliferator-associated receptor gamma


Sirtuin 1


Uncoupling protein 2



We would like to thank the Department of Physical Therapy (Congdon School of Health Sciences) and the School of Arts and Sciences for the use of shared lab space and equipment.

Author contribution

MGP made a significant contribution to the first draft of the manuscript and was lead author. MGP, MCS, LMG, and CJL assisted with experiments and manuscript preparation. TLG assisted with manuscript preparation. RAV designed and conducted experiments and assisted with manuscript preparation. MRK is corresponding author. He conceived the study, designed, conducted and oversaw all experiments, performed statistical analyses, oversaw MGP in drafting of the manuscript, and wrote subsequent versions of the manuscript as dictated by the reviewers. All authors read and approved the final manuscript.

Funding information

Financial support for this work was provided by the Department of Exercise Science within the Congdon School of Health Sciences.

Supplementary material

12192_2018_894_MOESM1_ESM.docx (278 kb)
Supplementary Fig. 1 (DOCX 277 kb)
12192_2018_894_MOESM2_ESM.docx (35 kb)
Supplemental Table 1 (DOCX 34 kb)


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Copyright information

© Cell Stress Society International 2018

Authors and Affiliations

  • Meghan G. Patton
    • 1
  • Trevor L. Gillum
    • 2
  • Mandy C. Szymanski
    • 1
  • Lacey M. Gould
    • 1
  • Claire J. Lauterbach
    • 1
  • Roger A. Vaughan
    • 1
  • Matthew R. Kuennen
    • 1
  1. 1.Department of Exercise ScienceHigh Point UniversityHigh PointUSA
  2. 2.Department of KinesiologyCalifornia Baptist UniversityRiversideUSA

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