The intrinsic activating factors that induce transcription of HSP72 in skeletal muscle following exercise remain unclear. We hypothesized that the cytosolic Ca2+ transient that occurs with depolarization is a determinant. We utilized intact, single skeletal muscle fibers from Xenopus laevis to test the role of the cytosolic Ca2+ transient and several other exercise-related factors (fatigue, hypoxia, AMP-kinase, and cross-bridge cycling) on the activation of HSP72 transcription. HSP72 and HSP60 mRNA levels were assessed with real time quantitative PCR; cytosolic [Ca2+] ([Ca2+]cyt) was assessed with fura-2. Both fatiguing and non-fatiguing contractions resulted in a significant increase in HSP72 mRNA. As expected, peak [Ca2+]cyt remained tightly coupled with peak developed tension in contracting fibers. Pre-treatment with N-benzyl-p-toluene sulfonamide (BTS) resulted in depressed peak developed tension with stimulation, while peak [Ca2+]cyt remained largely unchanged from control values. Despite excitation-contraction uncoupling, BTS-treated fibers displayed a significant increase in HSP72 mRNA. Treatment of fibers with hypoxia (PO2 <3 mm Hg) or AMP-kinase activation had no effect on HSP72 mRNA levels. These results suggest that the intermittent cytosolic Ca2+ transient that occurs with skeletal muscle depolarization provides a sufficient activating stimulus for HSP72 transcription. Metabolic or mechanical factors associated with fatigue development and cross-bridge cycling likely play a more limited role.
- Copyright © 2015, Journal of Applied Physiology