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Dying myofibers in elderly mouse skeletal muscles are characterized by the appearance of dystrophin-encircled vacuoles

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Abstract

The age-related loss of skeletal muscle mass and strength (sarcopenia) is predominantly attributed to myofiber atrophy, however the role or existence of myofiber death is currently unclear. We recently discovered dysmorphic myofibers in normal elderly mice resembling those that characterize the Autophagic Vacuolar Myopathies, and speculated that they may be myofibers caught in the act of dying. Since these myofibers were identifiable by Dystrophin Encircled Vacuoles and invaginations with Intracellular Localization we coined the acronym DEVILs and aimed to determine their frequency, pathogenesis and correlation with myofiber loss. In whole transverse sections of young (1–6 month) and elderly (22–26 month) C57Bl/6j mouse muscles, DEVILated myofiber number correlated with myofiber loss, being increasingly prevalent in aged extensor digitorum longus (R = 0.7, p < 0.001) and soleus (R = 0.6, p = 0.004) muscles, whilst rare in myofiber loss resistant muscles (cleido- and sternomastoid). In a cell viability dye-exclusion test, 17 ± 14 % of DEVILated myofibers stained positive and were accompanied by immunoglobulin infiltration compared to 1 ± 1 % of normal myofibers (p = 0.029). Virtually all DEVILs were acid-phosphatase reactive but contained p62 immunoreactivity and periodic acid-Schiff stained plaques. Compared to normal myofibers, BNIP3 immunostaining in DEVILated myofibers was reduced, whilst MAP-LC3b was indifferent. Cleaved-caspase 3 immunoreactivity was marginally elevated in DEVILated myofibers, but unaccompanied by nuclear DNA fragmentation. DEVILated myofibers were also identified in elderly rat (24 month) and cadaveric human (78 years) muscles. We argue that DEVIL formation reflects a previously undescribed fibre death process via a mechanism involving autophagic dysfunction and that the process may represent our first direct insight into the mechanism by which myofibers are lost in old age.

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Acknowledgments

We thank Dr Jon Cornwall for providing access to human tissue samples. This work was supported by the Department of Physiology at the University of Otago. NL was the grateful recipient of a University of Otago Master’s Scholarship and Postgraduate Publishing Bursary.

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The authors declare that they have no competing interests to disclose.

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  1. Department of Physiology, Otago School of Medical Sciences, University of Otago, P.O. Box 913, Dunedin, New Zealand

    Navneet Lal & Philip Sheard

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  1. Navneet Lal
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  2. Philip Sheard
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Correspondence to Philip Sheard.

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Lal, N., Sheard, P. Dying myofibers in elderly mouse skeletal muscles are characterized by the appearance of dystrophin-encircled vacuoles. Biogerontology 16, 443–459 (2015). https://doi.org/10.1007/s10522-015-9565-0

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