Abstract
Skeletal muscle of the dystrophin-deficient mdx mouse is hypersensitive to eccentric (ECC) contraction-induced strength loss due to plasmalemmal electrical dysfunction. Despite plasmalemmal inexcitability being a logical mechanism responsible for weakness, it remains unclear if processes up- and/or down-stream remain functionally intact in injured mdx muscle. The purpose of this study was to analyze additional processes necessary for excitation-contraction coupling that are potentially disrupted by ECC contractions. Anterior crural muscles (tibialis anterior, extensor digitorum longus [EDL], and extensor hallucis muscles) of wildtype (WT) and mdx mice were injured in vivo with 50 ECC contractions and torque was measured immediately before and after the contraction bout. Following the in vivo assessment, EDL ex vivo isometric and caffeine forces were analyzed. In vivo isometric torque and ex vivo force in WT muscle were reduced 38 and 30% (p < 0.001), while caffeine force was also reduced (p = 0.021), albeit to a lesser degree (9%). In contrast, in vivo isometric torque, ex vivo isometric force and ex vivo caffeine-induced force were all reduced 56–67% (p < 0.001) in mdx muscle and did not differ from one another (p = 0.114). Disproportional reductions in isometric strength and caffeine-induced force confirm that ECC contractions uncoupled the plasmalemma from the ryanodine receptors (RyRs) in WT muscle. In mdx muscle, the proportional reductions in isometric strength and caffeine-induced force following ECC contractions reveal that dysfunction occurs at and/or distal to the RyRs immediately post-injury. Thus, weakness in injured mdx muscle cannot be isolated to one mechanism, rather several steps of muscle contraction are disrupted.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.
References
Baumann CW, Rogers RG, Gahlot N, Ingalls CP (2014) Eccentric contractions disrupt FKBP12 content in mouse skeletal muscle. Physiol Rep. https://doi.org/10.14814/phy2.12081
Baumann CW, Rogers RG, Otis JS (2016) Utility of 17-(allylamino)-17-demethoxygeldanamycin treatment for skeletal muscle injury. Cell Stress Chaperones. https://doi.org/10.1007/s12192-016-0717-1
Baumann CW, Rogers RG, Otis JS, Ingalls CP (2016) Recovery of strength is dependent on mTORC1 signaling after eccentric muscle injury. Muscle Nerve. https://doi.org/10.1002/mus.25121
Baumann CW, Warren GL, Lowe DA (2020) Plasmalemma function is rapidly restored in mdx muscle after eccentric contractions. Med Sci Sports Exer 52:354–361. https://doi.org/10.1249/MSS.0000000000002126
Baumann CW, Lindsay A, Sidky SR et al (2021) Contraction-induced loss of plasmalemmal electrophysiological function is dependent on the dystrophin glycoprotein complex. Front Physiol. https://doi.org/10.3389/fphys.2021.757121
Bell CD, Conen PE (1968) Histopathological changes in Duchenne muscular dystrophy. J Neurol Sci 7:529–544. https://doi.org/10.1016/0022-510X(68)90058-0
Bellinger AM, Reiken S, Carlson C et al (2009) Hypernitrosylated ryanodine receptor calcium release channels are leaky in dystrophic muscle. Nat Med 15:325–330. https://doi.org/10.1038/nm.1916
Blaauw B, Agatea L, Toniolo L et al (2010) Eccentric contractions lead to myofibrillar dysfunction in muscular dystrophy. J Appl Physiol 108:105–111. https://doi.org/10.1152/japplphysiol.00803.2009
Brooks S (1998) Rapid recovery following contraction-induced injury to in situ skeletal muscles in mdx mice. J Muscle Res Cell Motil 19:179–187
Cairns S, Flatman J, Clausen T (1995) Relation between extracellular [K+], membrane potential and contraction in rat soleus muscle: modulation by the Na+-K + pump. Pflug Arch 430:909–915
Cairns SP, Hing WA, Slack JR et al (1997) Different effects of raised [K+]o on membrane potential and contraction in mouse fast- and slow-twitch muscle. Am J Physiol 273:C598–611. https://doi.org/10.1152/ajpcell.1997.273.2.C598
Call JA, Eckhoff MD, Baltgalvis KA et al (2011) Adaptive strength gains in dystrophic muscle exposed to repeated bouts of eccentric contraction. J Appl Physiol 111:1768–1777. https://doi.org/10.1152/japplphysiol.00942.2011
Call JA, Warren GL, Verma M, Lowe DA (2013) Acute failure of action potential conduction in mdx muscle reveals new mechanism of contraction-induced force loss. J Physiol 591:3765–3776. https://doi.org/10.1113/jphysiol.2013.254656
Corona BT, Balog EM, Doyle JA et al (2010) Junctophilin damage contributes to early strength deficits and EC coupling failure after eccentric contractions. Am J Physiol Physiol 298:C365–C376. https://doi.org/10.1152/ajpcell.00365.2009
Dangain J, Vrbova G (1984) Muscle development in mdx mutant mice. Muscle Nerve 7:700–704. https://doi.org/10.1002/MUS.880070903
Edwards RH, Hill DK, Jones DA, Merton PA (1977) Fatigue of long duration in human skeletal muscle after exercise. J Physiol 272:769–778
Emery AEH, Muntoni F, Quinlivan RCM (2015) Duchenne muscular dystrophy. Oxford University Press, Oxford
Grange R, Gainer T, Marschner K et al (2002) Fast-twitch skeletal muscles of dystrophic mouse pups are resistant to injury from acute mechanical stress. Am J Physiol Cell Physiol. https://doi.org/10.1152/AJPCELL.00450.2001
Head S, Williams D, Stephenson D (1992) Abnormalities in structure and function of limb skeletal muscle fibres of dystrophic mdx mice. Proc Biol Sci 248:163–169. https://doi.org/10.1098/RSPB.1992.0058
Herrmann F, Lüttgau H, Stephenson D (1999) Caffeine and excitation-contraction coupling in skeletal muscle: a stimulating story. J Muscle Res Cell Motil 20:223–236. https://doi.org/10.1023/A:1005496708505
Ingalls CP, Warren GL, Williams JH et al (1998) E-C coupling failure in mouse EDL muscle after in vivo eccentric contractions. J Appl Physiol 85:58–67. https://doi.org/10.1152/jappl.1998.85.1.58
Ingalls CP, Warren GL, Zhang J-Z et al (2004a) Dihydropyridine and ryanodine receptor binding after eccentric contractions in mouse skeletal muscle. J Appl Physiol 96:1619–1625. https://doi.org/10.1152/japplphysiol.00084.2003
Ingalls CP, Wenke JC, Nofal T, Armstrong RB (2004b) Adaptation to lengthening contraction-induced injury in mouse muscle. J Appl Physiol 97:1067–1076. https://doi.org/10.1152/japplphysiol.01058.2003
Jones DA, Howell S, Roussos C, Edwards RH (1982) Low-frequency fatigue in isolated skeletal muscles and the effects of methylxanthines. Clin Sci (Lond) 63:161–167
Kinali M, Arechavala-Gomeza V, Cirak S et al (2011) Muscle histology vs MRI in Duchenne muscular dystrophy. Neurology 76:346–353. https://doi.org/10.1212/WNL.0b013e318208811f
Kiriaev KL, Morley J et al (2021) Dystrophin-negative slow-twitch soleus muscles are not susceptible to eccentric contraction induced injury over the lifespan of the mdx mouse. Am J Physiol Cell Physiol. https://doi.org/10.1152/AJPCELL.00122.2021
Lindsay A, Southern WM, McCourt PM et al (2019) Variable cytoplasmic actin expression impacts the sensitivity of different dystrophin-deficient mdx skeletal muscle to eccentric contraction. FEBS J 286:2562–2576
Lindsay A, Baumann CW, Rebbeck RT et al (2020) Mechanical factors tune the sensitivity of mdx muscle to eccentric strength loss and its protection by antioxidant and calcium modulators. Skelet Muscle 10:3. https://doi.org/10.1186/s13395-020-0221-2
Liu M, Yue Y, Harper S et al (2005) Adeno-associated virus-mediated microdystrophin expression protects young mdx muscle from contraction-induced injury. Mol Ther 11:245–256. https://doi.org/10.1016/J.YMTHE.2004.09.013
Lowe DA, Warren GL, Ingalls CP et al (1995) Muscle function and protein metabolism after initiation of eccentric contraction-induced injury. J Appl Physiol 79:1260–1270. https://doi.org/10.1152/jappl.1995.79.4.1260
Lynch GS, Hinkle RT, Chamberlain JS et al (2001) Force and power output of fast and slow skeletal muscles from mdx mice 6–28 months old. J Physiol 535:591. https://doi.org/10.1111/J.1469-7793.2001.00591.X
Mázala D, Pratt S, Chen D et al (2015) SERCA1 overexpression minimizes skeletal muscle damage in dystrophic mouse models. Am J Physiol Cell Physiol 308:C699–C709. https://doi.org/10.1152/AJPCELL.00341.2014
McGreevy J, Hakim C, McIntosh M, Duan D (2015) Animal models of Duchenne muscular dystrophy: from basic mechanisms to gene therapy. Dis Model Mech 8:195–213. https://doi.org/10.1242/DMM.018424
Moens P, Baatsen PH, Maréchal G (1993) Increased susceptibility of EDL muscles from mdx mice to damage induced by contractions with stretch. J Muscle Res Cell Motil 14:446–451
Morine K, Sleeper M, Barton E, Sweeney H (2010) Overexpression of SERCA1a in the mdx diaphragm reduces susceptibility to contraction-induced damage. Hum Gene Ther 21:1735–1739. https://doi.org/10.1089/HUM.2010.077
Ng R, Metzger J, Claflin D, Faulkner J (2008) Poloxamer 188 reduces the contraction-induced force decline in lumbrical muscles from mdx mice. Am J Physiol Cell Physiol. https://doi.org/10.1152/AJPCELL.00017.2008
Petrof BJ, Shrager JB, Stedman HH et al (1993) Dystrophin protects the sarcolemma from stresses developed during muscle contraction. Proc Natl Acad Sci U S A 90:3710–3714
Pratt SJP, Shah SB, Ward CW et al (2013) Effects of in vivo injury on the neuromuscular junction in healthy and dystrophic muscles. J Physiol 591:559–570. https://doi.org/10.1113/jphysiol.2012.241679
Pratt SJP, Shah SB, Ward CW et al (2015) Recovery of altered neuromuscular junction morphology and muscle function in mdx mice after injury. Cell Mol Life Sci 72:153–164. https://doi.org/10.1007/s00018-014-1663-7
Renaud JM, Light P (1992) Effects of K + on the twitch and tetanic contraction in the sartorius muscle of the frog, Rana pipiens. Implication for fatigue in vivo. Can J Physiol Pharmacol 70:1236–1246
Roy P, Rau F, Ochala J et al (2016) Dystrophin restoration therapy improves both the reduced excitability and the force drop induced by lengthening contractions in dystrophic mdx skeletal muscle. Skelet Muscle 6:23. https://doi.org/10.1186/s13395-016-0096-4
Sharp P, Bye-a-Jee H, Wells D (2011) Physiological characterization of muscle strength with variable levels of dystrophin restoration in mdx mice following local antisense therapy. Mol Ther 19:165–171. https://doi.org/10.1038/MT.2010.213
Sonnemann K, Heun-Johnson H, Turner A et al (2009) Functional substitution by TAT-utrophin in dystrophin-deficient mice. PLoS Med. https://doi.org/10.1371/JOURNAL.PMED.61000083
Tanabe Y, Esaki K, Nomura T (1986) Skeletal muscle pathology in X chromosome-linked muscular dystrophy (mdx) mouse. Acta Neuropathol 69:91–95. https://doi.org/10.1007/BF00687043
Vohra RS, Lott D, Mathur S et al (2015) Magnetic resonance assessment of hypertrophic and pseudo-hypertrophic changes in lower leg muscles of boys with duchenne muscular dystrophy and their relationship to functional measurements. PLoS ONE. https://doi.org/10.1371/journal.pone.0128915
Warren GL, Lowe DA, Hayes DA et al (1993) Excitation failure in eccentric contraction-induced injury of mouse soleus muscle. J Physiol 468:487–499. https://doi.org/10.1113/jphysiol.1993.sp019783
Warren GL, Ingalls CP, Shah SJ, Armstrong RBB (1999) Uncoupling of in vivo torque production from EMG in mouse muscles injured by eccentric contractions. J Physiol 515:609
Warren GL, Hermann KM, Ingalls CP et al (2000) Decreased EMG median frequency during a second bout of eccentric contractions. Med Sci Sports Exerc 32:820–829
Warren GL, Ingalls CP, Lowe DA, Armstrong RB (2001) Excitation-contraction uncoupling: major role in contraction-induced muscle injury. Exerc Sport Sci Rev 29:82–87. https://doi.org/10.1097/00003677-200104000-00008
Willmann R, Possekel S, Dubach-Powell J et al (2009) Mammalian animal models for Duchenne muscular dystrophy. Neuromuscul Disord 19:241–249. https://doi.org/10.1016/j.nmd.2008.11.015
Acknowledgements
This work was funded by a Research Endowment from the American College of Sports Medicine Foundation (to C.W.B), a grant from the University of Minnesota Bob Allison Ataxia Research Center (to D. A. L.), and C.W.B was supported by the National Institutes of Health (T32-AG029796 & T32-AR007612).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Baumann, C.W., Ingalls, C.P. & Lowe, D.A. Mechanisms of weakness in Mdx muscle following in vivo eccentric contractions. J Muscle Res Cell Motil 43, 63–72 (2022). https://doi.org/10.1007/s10974-022-09617-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10974-022-09617-1