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Regulation of autophagic and mitophagic flux during chronic contractile activity-induced muscle adaptations

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Abstract

Autophagy and mitophagy are important for training-inducible muscle adaptations, yet it remains unclear how these systems are regulated throughout the adaptation process. Here, we studied autophagic and mitophagic flux in the skeletal muscles of Sprague–Dawley rats (300–500 g) exposed to chronic contractile activity (CCA; 3 h/day, 9 V, 10 Hz continuous, 0.1 ms pulse duration) for 1, 2, 5, and 7 days (N = 6–8/group). In order to determine the flux rates, colchicine (COL; 0.4 mg/ml/kg) was injected 48 h before tissue collection, and we evaluated differences of autophagosomal protein abundances (LC3-II and p62) between colchicine- and saline-injected animals. We confirmed that CCA resulted in mitochondrial adaptations, including improved state 3 respiration as early as day 1 in permeabilized muscle fibers, as well significant increases in mitochondrial respiratory capacity and marker proteins in IMF mitochondria by day 7. Mitophagic and autophagic flux (LC3-II and p62) were significantly decreased in skeletal muscle following 7 days of CCA. Notably, the mitophagic system seemed to be downregulated prior (day 3–5) to changes in autophagic flux (day 7), suggesting enhanced sensitivity of mitophagy compared to autophagy with chronic muscle contraction. Although we detected no significant change in the nuclear translocation of TFEB, a regulator of lysosomal biogenesis, CCA increased total TFEB protein, as well as LAMP1, in skeletal muscle. Thus, chronic muscle activity reduces mitophagy in parallel with improved mitochondrial function, and this is supported by enhanced lysosomal degradation capacity.

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Acknowledgements

We thank Jenny Lai for her dedicated technical assistance on this project.

Funding

This work was supported by funding from the Natural Sciences and Engineering Research Council of Canada (NSERC) to D. A. Hood. D. A. Hood is also the holder of a Canada Research Chair in Cell Physiology.

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Author notes

  1. Yuho Kim

    Present address: National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, 20892, USA

Authors and Affiliations

  1. Muscle Health Research Centre, Toronto, ON, Canada

    Yuho Kim, Matthew Triolo, Avigail T. Erlich & David A. Hood

  2. School of Kinesiology and Health Science, York University, Toronto, ON, M3J 1P3, Canada

    Yuho Kim, Matthew Triolo, Avigail T. Erlich & David A. Hood

Authors
  1. Yuho Kim
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  2. Matthew Triolo
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Corresponding author

Correspondence to David A. Hood.

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All animal procedures were approved and performed in accordance with the guidelines of the York University Animal Care Committee.

Additional information

This article is part of the special issue on Exercise Physiology: future opportunities and challenges in Pflügers Archiv—European Journal of Physiology

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Kim, Y., Triolo, M., Erlich, A.T. et al. Regulation of autophagic and mitophagic flux during chronic contractile activity-induced muscle adaptations. Pflugers Arch - Eur J Physiol 471, 431–440 (2019). https://doi.org/10.1007/s00424-018-2225-x

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  • DOI: https://doi.org/10.1007/s00424-018-2225-x

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