Abstract
It is well-known that denervation of motor nerves induces atrophy and decreases contractile force of the skeletal muscle. However, it is not completely understood how denervation alters calcium handling in the skeletal muscle. We investigated the effect of denervation on the expression and function of proteins involved in calcium handling. Two weeks after denervation of the right sciatic nerve in mice, we observed a significant decrease in mass and cross-sectional area of the ipsilateral tibialis anterior (TA) and flexor digitorum brevis (FDB) muscles. Also, we observed a significant decrease in the specific tetanus contractile force in the ipsilateral TA muscle. Calcium imaging of the ipsilateral FDB showed that the peak twitch and tetanus calcium concentrations were significantly decreased due to a decrease in calcium content of the sarcoplasmic reticulum (SR). Denervation reduced the maximum rate of sarcoplasmic/endoplasmic calcium ATPase (SERCA) activity. Interestingly, the amount of phospholamban (PLN), but not its transcripts, was increased in the ipsilateral vs. contralateral side after denervation, suggesting that denervation impairs constitutive regulation of PLN. Immunohistochemical analysis revealed increased PLN in all major fiber types in TA with IIx fibers showing a threefold higher expression than the contralateral side. These results suggest that the abnormal increase in PLN in the ipsilateral fast-twitch fibers may be involved in decreased SERCA activity, SR calcium content, peak calcium transients, and contractile forces of denervated muscles.
Similar content being viewed by others
References
Allard B (2018) From excitation to intracellular Ca2+ movements in skeletal muscle: Basic aspects and related clinical disorders. Neuromuscul Disord 28:394–401
Asahi M, Kurzydlowski K, Tada M, MacLennan DH (2002) Sarcolipin inhibits polymerization of phospholamban to induce superinhibition of sarco(endo)plasmic reticulum Ca2+-ATPases (SERCAs). J Biol Chem 277:26725–26728
Bhupathy P, Babu GJ, Periasamy M (2007) Sarcolipin and phospholamban as regulators of cardiac sarcoplasmic reticulum Ca2+ ATPase. J Mol Cell Cardiol 42:903–911
Bloemberg D, Quadrilatero J (2012) Rapid determination of myosin heavy chain expression in rat, mouse, and human skeletal muscle using multicolor immunofluorescence analysis. PLoS One 7:e35273
Brini M, Carafoli E (2009) Calcium pumps in health and disease. Physiol Rev 89:1341–1378
Brody IA (1966) Relaxing factor in denervated muscle: A possible explanation for fibrillations. Am J Physiol 211:1277–1280
Cosgrove BD, Gilbert PM, Porpiglia E et al (2014) Rejuvenation of the muscle stem cell population restores strength to injured aged muscles. Nat Med 20:255–264
Divet A, Lompré A-M, Huchet-Cadiou C (2005) Effect of cyclopiazonic acid, an inhibitor of the sarcoplasmic reticulum Ca-ATPase, on skeletal muscles from normal and mdx mice. Acta Physiol Scand 184:173–186
Dufresne SS, Dumont NA, Boulanger-Piette A et al (2016) Muscle RANK is a key regulator of Ca2+ storage, SERCA activity, and function of fast-twitch skeletal muscles. Am J Physiol Cell Physiol 310:C663–C672
Duhamel TA, Green HJ, Stewart RD et al (2007) Muscle metabolic, SR Ca2+ -cycling responses to prolonged cycling, with and without glucose supplementation. J Appl Physiol 103:1986–1998
Fajardo VA, Bombardier E, Vigna C et al (2013) Co-expression of SERCA isoforms, phospholamban and sarcolipin in human skeletal muscle fibers. PLoS One 8:e84304
Fajardo VA, Bombardier E, McMillan E et al (2015) Phospholamban overexpression in mice causes a centronuclear myopathy-like phenotype. Dis Model Mech 8:999–1009
Germinario E, Esposito A, Megighian A et al (2002) Early changes of type 2B fibers after denervation of rat EDL skeletal muscle. J Appl Physiol 92:2045–2052
Gonzalez-Freire M, de Cabo R, Studenski SA, Ferrucci L (2014) The neuromuscular junction: aging at the crossroad between nerves and muscle. Front Aging Neurosci 6:208
Harrer JM, Ponniah S, Ferguson DG, Kranias EG (1995) Expression of phospholamban in C2C12 cells and regulation of endogenous SERCA1 activity. Mol Cell Biochem 146:13–21
Huey KA, Bodine SC (1998) Changes in myosin mRNA and protein expression in denervated rat soleus and tibialis anterior. Eur J Biochem 256:45–50
Jorgensen AO, Jones LR (1986) Localization of phospholamban in slow but not fast canine skeletal muscle fibers. An immunocytochemical and biochemical study. J Biol Chem 261:3775–3781
Kho C, Lee A, Hajjar RJ (2012) Altered sarcoplasmic reticulum calcium cycling–targets for heart failure therapy. Nat Rev Cardiol 9:717–733
Kranias EG, Hajjar RJ (2012) Modulation of cardiac contractility by the phospholamban/SERCA2a regulatome. Circ Res 110:1646–1660
Leeuw T, Kapp M, Pette D (1994) Role of innervation for development and maintenance of troponin subunit isoform patterns in fast- and slow-twitch muscles of the rabbit. Differentiation 55:193–201
Loirat MJ, Lucas-Heron B, Ollivier B, Léoty C (1988) Calcium binding protein changes of sarcoplasmic reticulum from rat denervated skeletal muscle. Biosci Rep 8:369–378
Loy RE, Orynbayev M, Xu L et al (2011) Muscle weakness in Ryr1I4895T/WT knock-in mice as a result of reduced ryanodine receptor Ca2+ ion permeation and release from the sarcoplasmic reticulum. J Gen Physiol 137:43–57
Même W, Léoty C (2001) Cyclopiazonic acid and thapsigargin reduce Ca2+ influx in frog skeletal muscle fibres as a result of Ca2+ store depletion. Acta Physiol Scand 173:391–399
Midrio M (2006) The denervated muscle: Facts and hypotheses. A historical review. Eur J Appl Physiol 98:1–21
Midrio M, Danieli-Betto D, Megighian A, Betto R (1997) Early effects of denervation on sarcoplasmic reticulum properties of slow-twitch rat muscle fibres. Pflugers Arch 434:398–405
Murphy RM, Larkins NT, Mollica JP et al (2009) Calsequestrin content and SERCA determine normal and maximal Ca2+ storage levels in sarcoplasmic reticulum of fast- and slow-twitch fibres of rat. J Physiol (Lond) 587:443–460
Nakagawa T, Yokoe S, Asahi M (2016) Phospholamban degradation is induced by phosphorylation-mediated ubiquitination and inhibited by interaction with cardiac type Sarco(endo)plasmic reticulum Ca2+-ATPase. Biochem Biophys Res Commun 472:523–530
Palexas GN, Savage N, Isaacs H (1981) Characteristics of sarcoplasmic reticulum from normal and denervated rat skeletal muscle. Biochem J 200:11–15
Park KHJ (2015) Mechanisms of muscle denervation in aging: Insights from a mouse model of amyotrophic lateral sclerosis. Aging Dis 6:380–389
Péréon Y, Sorrentino V, Dettbarn C et al (1997) Dihydropyridine receptor and ryanodine receptor gene expression in long-term denervated rat muscles. 240:612–617
Periasamy M, Kalyanasundaram A (2007) SERCA pump isoforms: their role in calcium transport and disease. Muscle Nerve 35:430–442
Raffaello A, Laveder P, Romualdi C et al (2006) Denervation in murine fast-twitch muscle: short-term physiological changes and temporal expression profiling. Physiol Genomics 25:60–74
Rana ZA, Gundersen K, Buonanno A et al (2005) Imaging transcription in vivo: distinct regulatory effects of fast and slow activity patterns on promoter elements from vertebrate troponin I isoform genes. J Physiol 562:815–828
Rudolf R, Khan MM, Labeit S, Deschenes MR (2014) Degeneration of neuromuscular junction in age and dystrophy. Front Aging Neurosci 6:99
Salvatori S, Damiani E, Zorzato F et al (1988) Denervation-induced proliferative changes of triads in rabbit skeletal muscle. Muscle Nerve 11:1246–1259
Schiaffino S, Reggiani C (2011) Fiber types in mammalian skeletal muscles. Physiol Rev 91:1447–1531
Schmidt AG, Edes I, Kranias EG (2001) Phospholamban: a promising therapeutic target in heart failure? Cardiovasc Drugs Ther 15:387–396
Schulte L, Peters D, Taylor J et al (1994) Sarcoplasmic reticulum Ca2+ pump expression in denervated skeletal muscle. Am J Physiol 267:C617–C622
Slack JP, Grupp IL, Ferguson DG et al (1997) Ectopic expression of phospholamban in fast-twitch skeletal muscle alters sarcoplasmic reticulum Ca2+ transport and muscle relaxation. J Biol Chem 272:18862–18868
Song Q, Young KB, Chu G, Gulick J, Gerst M, Grupp IL, Robbins J, Kranias EG (2004) Overexpression of phospholamban in slow-twitch skeletal muscle is associated with depressed contractile function and muscle remodeling. FASEB J 18(9):974–976
Teng ACT, Miyake T, Yokoe S et al (2015) Metformin increases degradation of phospholamban via autophagy in cardiomyocytes. Proc Natl Acad Sci USA 112:7165–7170
Yokoe S, Asahi M (2017) Phospholamban Is Downregulated by pVHL-Mediated Degradation through Oxidative Stress in Failing Heart. Int J Mol Sci 18:2232
Zhao G, Li T, Brochet DXP et al (2015) STIM1 enhances SR Ca2+ content through binding phospholamban in rat ventricular myocytes. Proc Natl Acad Sci USA 112:E4792–E4801
Acknowledgements
We are grateful to Ms. Reiko Sakai for her secretarial assistance. This work was supported by Grants-in-aid for Scientific Research 25670641 (to HK) and 16K20046 (to MK) from the Ministry of Education, Culture, Sport, Science and Technology of Japan (MEXT) and a grant (to TN) from Shinshu Public Utility Foundation for Promotion of Medical Sciences, and by a research grant 41290769 from MSDKK (Tokyo, Japan) to MY.
Author information
Authors and Affiliations
Corresponding authors
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
Komatsu, M., Nakada, T., Kawagishi, H. et al. Increase in phospholamban content in mouse skeletal muscle after denervation. J Muscle Res Cell Motil 39, 163–173 (2018). https://doi.org/10.1007/s10974-019-09504-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10974-019-09504-2