Effects of PGC-1α overexpression on the myogenic response during skeletal muscle regeneration

Abstract

The ability of skeletal muscle to regenerate from injury is crucial for locomotion, metabolic health, and quality of life. Peroxisome proliferator-activated receptor-γ coactivator-1α (PGC1A) is a transcriptional coactivator required for mitochondrial biogenesis. Increased mitochondrial biogenesis is associated with improved muscle cell differentiation, however PGC1A's role in skeletal muscle regeneration following damage requires further investigation. The purpose of this study was to investigate the role of skeletal muscle-specific PGC1A overexpression during regeneration following damage. 22 C57BL/6J (WT) and 26 PGC1A muscle transgenic (A1) mice were injected with either phosphate-buffered saline (PBS, uninjured control) or Bupivacaine (MAR, injured) into their tibialis anterior (TA) muscle to induce skeletal muscle damage. TA muscles were extracted 3- or 28-days post-injury and analyzed for markers of regenerative myogenesis and protein turnover. Pgc1a mRNA was ∼10-20 fold greater in A1 mice. Markers of protein synthesis, AKT and 4EBP1, displayed decreases in A1 mice compared to WT at both timepoints indicating a decreased protein synthetic response. Myod mRNA was ∼75% lower compared to WT 3 days post-injection. WT mice exhibited decreased cross-sectional area of the TA muscle at 28 days post-injection with bupivacaine compared to all other groups. PGC1A overexpression modifies the myogenic response during regeneration.

Keywords

PGC-1α / Skeletal muscle / Muscle regeneration / Satellite cells / Protein turnover / p38 MAPK

References

[[1]] WR Frontera, J Ochala. Skeletal muscle: a brief review of structure and function. Calcif Tissue Int, 96 (3) ( 2015), pp. 183-195, DOI: 10.1007/s00223-014-9915-y

[[2]] M Tieland, I Trouwborst, BC Clark. Skeletal muscle performance and ageing. J Cachexia Sarcopenia Muscle, 9 (1) ( 2018), pp. 3-19, DOI: 10.1002/jcsm.12238

[[3]] M Flück, H Hoppeler. Molecular basis of skeletal muscle plasticity--from gene to form and function. Rev Physiol Biochem Pharmacol, 146 ( 2003), pp. 159-216, DOI: 10.1007/s10254-002-0004-7

[[4]] S Orimo, E Hiyamuta, K Arahata, H Sugita. Analysis of inflammatory cells and complement C3 in bupivacaine-induced myonecrosis. Muscle Nerve, 14 (6) ( 1991), pp. 515-520, DOI: 10.1002/mus.880140605

[[5]] JG Tidball, E Berchenko, J Frenette. Macrophage invasion does not contribute to muscle membrane injury during inflammation. J Leukoc Biol, 65 (4) ( 1999), pp. 492-498.

[[6]] GL Warren, M Summan, X Gao, R Chapman, T Hulderman, PP Simeonova. Mechanisms of skeletal muscle injury and repair revealed by gene expression studies in mouse models. J Physiol, 582 (Pt 2) ( 2007), pp. 825-841, DOI: 10.1113/jphysiol.2007.132373

[[7]] Z Yan, S Choi, X Liu, et al.. Highly coordinated gene regulation in mouse skeletal muscle regeneration. J Biol Chem, 278 (10) ( 2003), pp. 8826-8836, DOI: 10.1074/jbc.M209879200

[[8]] Y Saito, I Nonaka. Initiation of satellite cell replication in bupivacaine-induced myonecrosis. Acta Neuropathol, 88 (3) ( 1994), pp. 252-257

[[9]] MH Brooke, GM Fenichel, RC Griggs, et al.. Duchenne muscular dystrophy: patterns of clinical progression and effects of supportive therapy. Neurology, 39 (4) ( 1989), pp. 475-481, DOI: 10.1212/wnl.39.4.475

[[10]] AE Emery. Duchenne muscular dystrophy--Meryon’s disease. Neuromuscul Disord, 3 (4) ( 1993), pp. 263-266, DOI: 10.1016/0960-8966(93)90018-f

[[11]] R Koopman, CH Ly, JG Ryall.A metabolic link to skeletal muscle wasting and regeneration. Front Physiol, 5 ( 2014), p. 32, DOI: 10.3389/fphys.2014.00032

[[12]] TA Washington, L Brown, DA Smith, G Davis, J Baum, W Bottje. Monocarboxylate transporter expression at the onset of skeletal muscle regeneration. Phys Rep, 1 (4) ( 2013), Article e00075, DOI: 10.1002/phy2.75

[[13]] F Pala, D Di Girolamo, S Mella, et al.. Distinct metabolic states govern skeletal muscle stem cell fates during prenatal and postnatal myogenesis. J Cell Sci, 131 (14) ( 2018), p. jcs212977, DOI: 10.1242/jcs.212977

[[14]] S Dell’Orso, AH Juan, KD Ko, et al.. Single cell analysis of adult mouse skeletal muscle stem cells in homeostatic and regenerative conditions. Development, 146 (12) ( 2019), p. dev174177, DOI: 10.1242/dev.174177

[[15]] DW McKellar, LD Walter, LT Song, et al.. Large-scale integration of single-cell transcriptomic data captures transitional progenitor states in mouse skeletal muscle regeneration. Commun Biol, 4 (1) ( 2021), p. 1280, DOI: 10.1038/s42003-021-02810-x

[[16]] N Yanay, M Rabie, Y Nevo.Impaired regeneration in dystrophic muscle-new target for therapy. Front Mol Neurosci, 13 ( 2020), p. 69, DOI: 10.3389/fnmol.2020.00069

[[17]] A Rugowska, A Starosta, P Konieczny.Epigenetic modifications in muscle regeneration and progression of Duchenne muscular dystrophy. Clin Epigenet, 13 (1) ( 2021), p. 13, DOI: 10.1186/s13148-021-01001-z

[[18]] S Duguez, L Feasson, C Denis, D Freyssenet. Mitochondrial biogenesis during skeletal muscle regeneration. Am J Physiol Endocrinol Metab, 282 (4) ( 2002), pp. E802-E809, DOI: 10.1152/ajpendo.00343.2001

[[19]] A Wagatsuma, N Kotake, S Yamada. Muscle regeneration occurs to coincide with mitochondrial biogenesis. Mol Cell Biochem, 349 (1-2) ( 2011), pp. 139-147, DOI: 10.1007/s11010-010-0668-2

[[20]] C Kang, LL Ji. Role of PGC-1α signaling in skeletal muscle health and disease. Ann N Y Acad Sci, 1271 ( 2012), pp. 110-117, DOI: 10.1111/j.1749-6632.2012.06738.x

[[21]] J Lin, H Wu, PT Tarr, et al.. Transcriptional co-activator PGC-1 alpha drives the formation of slow-twitch muscle fibres. Nature, 418 (6899) ( 2002), pp. 797-801, DOI: 10.1038/nature00904

[[22]] Y Hatazawa, K Minami, R Yoshimura, et al.. Deletion of the transcriptional coactivator PGC1alpha in skeletal muscles is associated with reduced expression of genes related to oxidative muscle function. Biochem Biophys Res Commun, 481 (3-4) ( 2016), pp. 251-258, DOI: 10.1016/j.bbrc.2016.10.133

[[23]] PJ Adhihetty, G Uguccioni, L Leick, J Hidalgo, H Pilegaard, DA Hood. The role of PGC-1alpha on mitochondrial function and apoptotic susceptibility in muscle. Am J Physiol Cell Physiol, 297 (1) ( 2009), pp. C217-C225, DOI: 10.1152/ajpcell.00070.2009

[[24]] C Handschin, S Chin, P Li, et al.. Skeletal muscle fiber-type switching, exercise intolerance, and myopathy in PGC-1alpha muscle-specific knock-out animals. J Biol Chem, 282 (41) ( 2007), pp. 30014-30021, DOI: 10.1074/jbc.M704817200

[[25]] I Dinulovic, R Furrer, S Di Fulvio, A Ferry, M Beer, C Handschin.PGC-1α modulates necrosis, inflammatory response, and fibrotic tissue formation in injured skeletal muscle. Skeletal Muscle, 6 ( 2016), p. 38, DOI: 10.1186/s13395-016-0110-x

[[26]] NP Greene, DE Lee, JL Brown, et al.. Mitochondrial quality control, promoted by PGC-1α is dysregulated by Western diet-induced obesity and partially restored by moderate physical activity in mice. Phys Rep, 3 (7) ( 2015), DOI: 10.14814/phy2.12470

[[27]] RA Perry, LA Brown, DE Lee, et al.. Differential effects of leucine supplementation in young and aged mice at the onset of skeletal muscle regeneration. Mech Ageing Dev, 157 ( 2016), pp. 7-16, DOI: 10.1016/j.mad.2016.05.007

[[28]] LA Brown, DE Lee, JF Patton, et al.. Diet-induced obesity alters anabolic signalling in mice at the onset of skeletal muscle regeneration. Acta Physiol, 215 (1) ( 2015), pp. 46-57, DOI: 10.1111/apha.12537

[[29]] C van der Poel, LE Gosselin, JD Schertzer, et al.. Ageing prolongs inflammatory marker expression in regenerating rat skeletal muscles after injury. J Inflamm, 8 (1) ( 2011), p. 41, DOI: 10.1186/1476-9255-8-41

[[30]] NP Greene, MI Nilsson, TA Washington, et al.. Impaired exercise-induced mitochondrial biogenesis in the obese Zucker rat, despite PGC-1α induction, is due to compromised mitochondrial translation elongation. Am J Physiol Endocrinol Metab, 306 (5) ( 2014), pp. E503-E511, DOI: 10.1152/ajpendo.00671.2013

[[31]] TA Washington, JP White, JM Davis, et al.. Skeletal muscle mass recovery from atrophy in IL-6 knockout mice. Acta Physiol, 202 (4) ( 2011), pp. 657-669, DOI: 10.1111/j.1748-1716.2011.02281.x

[[32]] A Musaro. Muscle homeostasis and regeneration: from molecular mechanisms to therapeutic opportunities. Cells, 9 (9) ( 2020), p. 2033, DOI: 10.3390/cells9092033

[[33]] S Summermatter, G Santos, J Pérez-Schindler, C Handschin. Skeletal muscle PGC-1α controls whole-body lactate homeostasis through estrogen-related receptor α-dependent activation of LDH B and repression of LDH A. Proc Natl Acad Sci U S A, 110 (21) ( 2013), pp. 8738-8743, DOI: 10.1073/pnas.1212976110

[[34]] CR Benton, Y Yoshida, J Lally, XX Han, H Hatta, A Bonen. PGC-1alpha increases skeletal muscle lactate uptake by increasing the expression of MCT1 but not MCT2 or MCT4. Physiol Genom, 35 (1) ( 2008), pp. 45-54, DOI: 10.1152/physiolgenomics.90217.2008

[[35]] M Fan, J Rhee, J St-Pierre, et al.. Suppression of mitochondrial respiration through recruitment of p160 myb binding protein to PGC-1alpha: modulation by p38 MAPK. Genes Dev, 18 (3) ( 2004), pp. 278-289, DOI: 10.1101/gad.1152204

[[36]] GL Johnson, R Lapadat. Mitogen-activated protein kinase pathways mediated by ERK, JNK, and p38 protein kinases. Science, 298 (5600) ( 2002), pp. 1911-1912, DOI: 10.1126/science.1072682

[[37]] M Wang, PR Crisostomo, C Herring, KK Meldrum, DR Meldrum. Human progenitor cells from bone marrow or adipose tissue produce VEGF, HGF, and IGF-I in response to TNF by a p38 MAPK-dependent mechanism. Am J Physiol Regul Integr Comp Physiol, 291 (4) ( 2006), pp. R880-R884, DOI: 10.1152/ajpregu.00280.2006

[[38]] S Lim, KR Dunlap, ME Rosa-Caldwell, et al.. Comparative plasma proteomics in muscle atrophy during cancer-cachexia and disuse: the search for atrokines. Phys Rep, 8 (19) ( 2020), Article e14608, DOI: 10.14814/phy2.14608

[[39]] YP Li, Y Chen, J John, et al.. TNF-alpha acts via p 38 MAPK to stimulate expression of the ubiquitin ligase atrogin1/MAFbx in skeletal muscle. Faseb J, 19 (3) ( 2005), pp. 362-370, DOI: 10.1096/fj.04-2364com

[[40]] J Kim, KJ Won, HM Lee, et al.. p 38 MAPK participates in muscle-specific RING finger 1-mediated atrophy in cast-immobilized rat gastrocnemius muscle. KOREAN J PHYSIOL PHARMACOL, 13 (6) ( 2009), pp. 491-496, DOI: 10.4196/kjpp.2009.13.6.491

[[41]] DR Bolster, LS Jefferson, SR Kimball. Regulation of protein synthesis associated with skeletal muscle hypertrophy by insulin-, amino acid- and exercise-induced signalling. Proc Nutr Soc, 63 (2) ( 2004), pp. 351-356, DOI: 10.1079/PNS2004355

[[42]] DR Plant, FE Colarossi, GS Lynch. Notexin causes greater myotoxic damage and slower functional repair in mouse skeletal muscles than bupivacaine. Muscle Nerve, 34 (5) ( 2006), pp. 577-585, DOI: 10.1002/mus.20616

[[43]] E Perdiguero, D Cornelison.Erratum to: isolation, culture, and immunostaining of skeletal muscle myofibers from wildtype and nestin-GFP mice as a means to analyze satellite cells. Methods Mol Biol, 1556 ( 2017), p. C1, DOI: 10.1007/978-1-4939-6771-1_21

[[44]] K Guo, J Wang, V Andrés, RC Smith, K Walsh. MyoD-induced expression of p21 inhibits cyclin-dependent kinase activity upon myocyte terminal differentiation. Mol Cell Biol, 15 (7) ( 1995), pp. 3823-3829, DOI: 10.1128/mcb.15.7.3823

[[45]] O Halevy, BG Novitch, DB Spicer, et al.. Correlation of terminal cell cycle arrest of skeletal muscle with induction of p 21 by MyoD. Science, 267 (5200) ( 1995), pp. 1018-1021, DOI: 10.1126/science.7863327

[[46]] P Brien, D Pugazhendhi, S Woodhouse, D Oxley, JM Pell. p38α MAPK regulates adult muscle stem cell fate by restricting progenitor proliferation during postnatal growth and repair. Stem Cell, 31 (8) ( 2013), pp. 1597-1610, DOI: 10.1002/stem.1399

[[47]] E Perdiguero, V Ruiz-Bonilla, AL Serrano, P Muñoz-Cánoves. Genetic deficiency of p38alpha reveals its critical role in myoblast cell cycle exit: the p38alpha-JNK connection. Cell Cycle, 6 (11) ( 2007), pp. 1298-1303, DOI: 10.4161/cc.6.11.4315

[[48]] XZ Wang, D Ron. Stress-induced phosphorylation and activation of the transcription factor CHOP (GADD153) by p38 MAP Kinase. Science, 272 (5266) ( 1996), pp. 1347-1349, DOI: 10.1126/science.272.5266.1347

[[49]] J Alter, E Bengal. Stress-induced C/EBP homology protein (CHOP) represses MyoD transcription to delay myoblast differentiation. PLoS One, 6 (12) ( 2011), Article e29498, DOI: 10.1371/journal.pone.0029498

[[50]] N Ohoka, S Yoshii, T Hattori, K Onozaki, H Hayashi. TRB3, a novel ER stress-inducible gene, is induced via ATF4-CHOP pathway and is involved in cell death. EMBO J, 24 (6) ( 2005), pp. 1243-1255, DOI: 10.1038/sj.emboj.7600596

[[51]] S Kato, K Du. TRB 3 modulates C2C12 differentiation by interfering with Akt activation. Biochem Biophys Res Commun, 353 (4) ( 2007), pp. 933-938, DOI: 10.1016/j.bbrc.2006.12.161

[[52]] RH Choi, A McConahay, HW Jeong, et al.. Tribbles 3 regulates protein turnover in mouse skeletal muscle. Biochem Biophys Res Commun, 493 (3) ( 2017), pp. 1236-1242, DOI: 10.1016/j.bbrc.2017.09.134

[[53]] EJ Jones, PA Bishop, AK Woods, JM Green. Cross-sectional area and muscular strength: a brief review. Sports Med, 38 (12) ( 2008), pp. 987-994, DOI: 10.2165/00007256-200838120-00003

The authors would like to thank the Exercise Science Research Center staff at the University of Arkansas for all of their hard work and all administrative contributions towards this project.