bims-moremu 2021-06-27 papers

bims-moremu

Biomed News

on Molecular regulators of muscle mass

Issue of 2021–06–27
27 papers selected by
Anna Vainshtein, Craft Science Inc.

Regulation of Skeletal Muscle Satellite Cell Differentiation by Omega-3 Polyunsaturated Fatty Acids: A Critical Review.
Muscle-nerve communication and the molecular assessment of human skeletal muscle denervation with aging.
Nuclear factor E2-related factor 2 (NRF2) deficiency accelerates fast fibre type transition in soleus muscle during space flight.
Resveratrol improves muscle regeneration in obese mice through enhancing mitochondrial biogenesis.
Myofibre Hypertrophy in the Absence of Changes to Satellite Cell Content Following Concurrent Exercise Training in Young Healthy Men.
Vector-mediated expression of muscle specific kinase restores specific force to muscles in the mdx mouse model of Duchenne muscular dystrophy.
p38 MAPKs - roles in skeletal muscle physiology, disease mechanisms, and as potential therapeutic targets.
Skeletal muscle weakness in hypothyroidism.
C2C12 myoblasts are more sensitive to the toxic effects of simvastatin than myotubes and show impaired proliferation and myotube formation.
mRNA distribution in skeletal muscle is associated with mRNA size.
Muscle ankyrin repeat protein 1 (MARP1) locks titin to the sarcomeric thin filament and is a passive force regulator.
Gelsolin and dCryAB act downstream of muscle identity genes and contribute to preventing muscle splitting and branching in Drosophila.
The consequences of a high-calorie diet background before calorie restriction on skeletal muscles in a mouse model.
Hyperpolarized 13C MR Spectroscopy Depicts in Vivo Effect of Exercise on Pyruvate Metabolism in Human Skeletal Muscle.
Pathomechanisms and biomarkers in facioscapulohumeral muscular dystrophy: roles of DUX4 and PAX7.
Fibroblast fusion to the muscle fiber regulates myotendinous junction formation.
Effects of physical exercise in sarcopenia. A systematic review.
Anti-inflammatory nanoparticles significantly improve muscle function in a murine model of advanced muscular dystrophy.
Unexpected contribution of fibroblasts to muscle lineage as a mechanism for limb muscle patterning.
Plasma lipidomic analysis shows a disease progression signature in mdx mice.
Single-Cell RNAseq Analysis of lncRNAs.
Functional roles of non-coding RNAs in atrophy.
Development of metabolic dysfunction in mice lacking chemerin.
Muscle clock: how does muscle physiology change in 24 hours in response to exercise?
A MATLAB toolbox for muscle diffusion-tensor MRI tractography.
Morphological changes and functional circRNAs screening of rabbit skeletal muscle development.
Neural adaptations to long-term resistance training: evidence for the confounding effect of muscle size on the interpretation of surface electromyography.

Front Physiol. 2021 ;12 682091

Regulation of Skeletal Muscle Satellite Cell Differentiation by Omega-3 Polyunsaturated Fatty Acids: A Critical Review.

Peter O Isesele, Vera C Mazurak.

  Skeletal muscle is composed of multinuclear cells called myofibres, which are formed by the fusion of myoblasts during development. The size of the muscle fiber and mass of skeletal muscle are altered in response to several pathological and physiological conditions. Skeletal muscle regeneration is primarily mediated by muscle stem cells called satellite cells (SCs). In response to injury, these SCs replenish myogenic progenitor cells to form new myofibers to repair damaged muscle. During myogenesis, activated SCs proliferate and differentiate to myoblast and then fuse with one another to form muscle fibers. A reduced number of SCs and an inability to undergo myogenesis may contribute to skeletal muscle disorders such as atrophy, cachexia, and sarcopenia. Myogenic regulatory factors (MRF) are transcription factors that regulate myogenesis and determines whether SCs will be in the quiescent, activated, committed, or differentiated state. Mitochondria oxidative phosphorylation and oxidative stress play a role in the determination of the fate of SCs. The potential activation and function of SCs are also affected by inflammation during skeletal muscle regeneration. Omega-3 polyunsaturated fatty acids (PUFAs) show promise to reduce inflammation, maintain muscle mass during aging, and increase the functional capacity of the muscle. The aim of this critical review is to highlight the role of omega-3 PUFAs on the myogenic differentiation of SCs and pathways affected during the differentiation process, including mitochondrial function and inflammation from the current body of literature.

Keywords:  inflammation; myogenesis; omega-3; satellite cell; skeletal muscle

DOI:  https://doi.org/10.3389/fphys.2021.682091
Am J Physiol Cell Physiol. 2021 06 23.

Muscle-nerve communication and the molecular assessment of human skeletal muscle denervation with aging.

Casper Søndenbroe, Jesper L Andersen, Abigail L Mackey.

Muscle fiber denervation is a major contributor to the decline in physical function observed with aging. Denervation can occur through breakdown of the NMJ itself, affecting only that particular fiber, or through the death of a motoneuron, which can lead to a loss of all the muscle fibers in that motor unit. In this review we discuss the muscle-nerve relationship, where signaling from both the motor neuron and the muscle fiber is required for maximal preservation of neuromuscular function in old age. Physical activity is likely to be the most important single factor that can contribute to this preservation. Furthermore, we propose that inactivity is not an innocent bystander, but plays an active role in denervation through the production of signals hostile to neuron survival. Investigating denervation in human muscle tissue samples is challenging due to the shared protein profile of regenerating and denervated muscle fibers. In this review we provide a detailed overview of the key traits observed in immunohistochemical preparations of muscle biopsies from healthy young and elderly individuals. Overall, a combination of assessing tissue samples, circulating biomarkers, and electrophysiological assessments in humans will prove fruitful in the quest to gain more understanding of denervation of skeletal muscle. In addition, cell culture models represent a valuable tool in the search for key signaling factors exchanged between muscle and nerve, and which exercise has the capacity to alter.

DOI: https://doi.org/10.1152/ajpcell.00174.2021
Commun Biol. 2021 Jun 24. 4(1): 787

Nuclear factor E2-related factor 2 (NRF2) deficiency accelerates fast fibre type transition in soleus muscle during space flight.

Takuto Hayashi, Takashi Kudo, Ryo Fujita, Shin-Ichiro Fujita, Hirona Tsubouchi, Sayaka Fuseya, Riku Suzuki, Michito Hamada, Risa Okada, Masafumi Muratani, Dai Shiba, Takafumi Suzuki, Eiji Warabi, Masayuki Yamamoto, Satoru Takahashi.

Microgravity induces skeletal muscle atrophy, particularly in the soleus muscle, which is predominantly composed of slow-twitch myofibre (type I) and is sensitive to disuse. Muscle atrophy is commonly known to be associated with increased production of reactive oxygen species. However, the role of NRF2, a master regulator of antioxidative response, in skeletal muscle plasticity during microgravity-induced atrophy, is not known. To investigate the role of NRF2 in skeletal muscle within a microgravity environment, wild-type and Nrf2-knockout (KO) mice were housed in the International Space Station for 31 days. Gene expression and histological analyses demonstrated that, under microgravity conditions, the transition of type I (oxidative) muscle fibres to type IIa (glycolytic) was accelerated in Nrf2-KO mice without affecting skeletal muscle mass. Therefore, our results suggest that NRF2 affects myofibre type transition during space flight.

DOI: https://doi.org/10.1038/s42003-021-02334-4
J Nutr Biochem. 2021 Jun 22. pii: S0955-2863(21)00224-2. [Epub ahead of print] 108804

Resveratrol improves muscle regeneration in obese mice through enhancing mitochondrial biogenesis.

Wenjing Niu, Haibo Wang, Bo Wang, Xueying Mao, Min Du.

  Obesity is increasing rapidly worldwide and is accompanied by many complications, including impaired muscle regeneration. Obesity is known to inhibit AMP-activated protein kinase (AMPK) activity, which impedes mitochondrial biogenesis, myogenic differentiation and muscle regeneration. Resveratrol has an effective anti-obesity effect, but its effect on regeneration of muscle in obese mice remains to be tested. We hypothesized that resveratrol activates AMPK and mitochondrial biogenesis to improve muscle regeneration. Male C57BL/6J mice were fed a control diet or a 60% high-fat diet with or without resveratrol supplementation for 8 weeks and, then, the tibialis anterior muscle was subjected to cardiotoxin-induced muscle injury. Muscle tissue was collected at 3 and 7days after injury. We found that resveratrol enhanced both proliferation and differentiation of satellite cells following injury in obese mice. Markers of mitochondrial biogenesis were upregulated in resveratrol-treated mice. In C2C12 myogenic cells, resveratrol activated AMPK and stimulated the expression of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α), which were associated with enhanced myogenic differentiation. Such effects of resveratrol were abolished by AMPKα1 ablation, showing the mediatory roles of AMPK. In summary, dietary resveratrol activates AMPK/PGC-1α axis to facilitate mitochondrial biogenesis and muscle regeneration impaired due to obesity.

Keywords:  AMPK; mitochondrial biogenesis; muscle; obesity; regeneration; resveratrol

DOI:  https://doi.org/10.1016/j.jnutbio.2021.108804
Front Physiol. 2021 ;12 625044

Myofibre Hypertrophy in the Absence of Changes to Satellite Cell Content Following Concurrent Exercise Training in Young Healthy Men.

Baubak Shamim, Donny M Camera, Jamie Whitfield.

  Concurrent exercise training has been suggested to create an 'interference effect,' attenuating resistance training-based skeletal muscle adaptations, including myofibre hypertrophy. Satellite cells support myofibre hypertrophy and are influenced by exercise mode. To determine whether satellite cells contribute to the 'interference effect' changes in satellite cell and myonuclear content were assessed following a period of training in 32 recreationally active males (age: 25 ± 5 year; body mass index: 24 ± 3 kg⋅m-2; mean ± SD) who undertook 12-week of either isolated (3 d⋅w-1) resistance (RES; n = 10), endurance (END; n = 10), or alternate day (6 d⋅w-1) concurrent (CET, n = 12) training. Skeletal muscle biopsies were obtained pre-intervention and after 2, 8, and 12 weeks of training to determine fibre type-specific cross-sectional area (CSA), satellite cell content (Pax7+DAPI+), and myonuclei (DAPI+) using immunofluorescence microscopy. After 12 weeks, myofibre CSA increased in all training conditions in type II (P = 0.0149) and mixed fibres (P = 0.0102), with no difference between conditions. Satellite cell content remained unchanged after training in both type I and type II fibres. Significant correlations were observed between increases in fibre type-specific myonuclear content and CSA of Type I (r = 0.63, P < 0.0001), Type II (r = 0.69, P < 0.0001), and mixed fibres (r = 0.72, P < 0.0001). Resistance, endurance, and concurrent training induce similar myofibre hypertrophy in the absence of satellite cell and myonuclear pool expansion. These findings suggest that myonuclear accretion via satellite cell fusion is positively correlated with hypertrophy after 12 weeks of concurrent training, and that individuals with more myonuclear content displayed greater myofibre hypertrophy.

Keywords:  concurrent exercise; endurance exercise; resistance exercise; satellite cells; skeletal muscle

DOI:  https://doi.org/10.3389/fphys.2021.625044
Exp Physiol. 2021 Jun 10.

Vector-mediated expression of muscle specific kinase restores specific force to muscles in the mdx mouse model of Duchenne muscular dystrophy.

Joanne Ban, Besa Beqaj, William D Phillips.

   NEW FINDINGS: What is the central question of this study? The (dystrophin-deficient) muscles of mdx mice generate less contractile force per cross-sectional area (specific force) than those of healthy wild-type mice: what is the influence of muscle specific kinase (MuSK) upon the properties of the tibialis anterior (TA) muscle in mdx mice? What is the main finding and its importance? Injection of adeno-associated viral vector encoding MuSK into the TA muscle of young mdx mice increased the specific force of the muscle, suggesting the MuSK signalling system has the potential to restore healthy growth to dystrophin-deficient muscles.
ABSTRACT: In the mdx mouse model of Duchenne muscular dystrophy, muscle fibres are fragile and prone to injury and degeneration. Compared to wild-type mice, muscles of mdx mice also develop less specific force (contractile force/cross-sectional area). We recently reported that injecting adeno-associated viral vector encoding muscle specific kinase (AAV-MuSK) into muscles of mdx mice increased utrophin expression and made the muscles more resistant to acute stretch-induced injury. Here we injected AAV-MuSK unilaterally into the tibialis anterior muscle of mdx mice at a younger age (4 weeks), and recorded contraction force from the muscles in situ at 12 weeks of age. Compared to contralateral empty-vector control muscles, muscles injected with AAV-MuSK produced 28% greater specific force (P = 0.0005). They did not undergo the compensatory hypertrophy that normally occurs in muscles of mdx mice. Injection of AAV encoding rapsyn (a downstream effector of MuSK signalling) caused no such improvement in muscle strength. Muscles injected with AAV-MuSK displayed a 10% reduction in the number of fibres with centralized nuclei (P = 0.0015). Our results in mdx mice suggest that elevating the expression of MuSK can reduce the incidence of muscle fibre regeneration and improve the strength of dystrophin-deficient muscles.

Keywords:  MuSK; dystrophin; utrophin

DOI:  https://doi.org/10.1113/EP089439
JCI Insight. 2021 Jun 22. pii: 149915. [Epub ahead of print]6(12):

p38 MAPKs - roles in skeletal muscle physiology, disease mechanisms, and as potential therapeutic targets.

Christopher M Brennan, Charles P Emerson, Jane Owens, Nicolas Christoforou.

p38 MAPKs play a central role in orchestrating the cellular response to stress and inflammation and in the regulation of myogenesis. Potent inhibitors of p38 MAPKs have been pursued as potential therapies for several disease indications due to their antiinflammatory properties, although none have been approved to date. Here, we provide a brief overview of p38 MAPKs, including their role in regulating myogenesis and their association with disease progression. Finally, we discuss targeting p38 MAPKs as a therapeutic approach for treating facioscapulohumeral muscular dystrophy and other muscular dystrophies by addressing multiple pathological mechanisms in skeletal muscle.

DOI: https://doi.org/10.1172/jci.insight.149915
Nat Rev Endocrinol. 2021 Jun 22.

Skeletal muscle weakness in hypothyroidism.

Olivia Tysoe.

DOI: https://doi.org/10.1038/s41574-021-00528-8
Biochem Pharmacol. 2021 Jun 07. pii: S0006-2952(21)00262-8. [Epub ahead of print]190 114649

C2C12 myoblasts are more sensitive to the toxic effects of simvastatin than myotubes and show impaired proliferation and myotube formation.

Gerda M Sanvee, Jamal Bouitbir, Stephan Krähenbühl.

  Statins reduce cardiovascular complications in patients with high LDL-cholesterol but are associated with myopathy. We compared the toxicity of simvastatin of C2C12 myoblasts and myotubes. Since myoblasts can proliferate and fuse to myotubes, myoblasts can be considered as satellite cells and myotubes as mature muscle fibers. Simvastatin increased plasma membrane permeability and decreased the cellular ATP content in both myoblasts and myotubes, but with a stronger effect on myoblasts. While insulin prevented cytotoxicity up to 8 h after addition of simvastatin to myotubes, prevention in myoblasts required simultaneous addition. Mevalonate and geranylgeraniol prevented simvastatin-associated cytotoxicity in both myoblasts and myotubes. Simvastatin impaired the phosphorylation of the insulin receptor (IR β), Akt ser473 and S6rp, and increased phosphorylation of AMPK thr172 in both myotubes and myoblasts, which was prevented by insulin and mevalonate. Simvastatin impaired oxygen consumption and increased superoxide production by myoblasts and myotubes and induced apoptosis via cytochrome c release. In addition, simvastatin impaired proliferation and fusion of myoblasts to myotubes by inhibiting the expression of the nuclear transcription factor MyoD and of the metalloprotease ADAM-12. Decreased expression of the proliferation factor Ki-67 and of ADAM-12 were also observed in gastrocnemius of mice treated with simvastatin. In conclusion, myoblasts were more susceptible to the toxic effects of simvastatin and simvastatin impaired myoblast proliferation and myotube formation. Impaired muscle regeneration may represent a new mechanism of statin myotoxicity.

Keywords:  C2C12 myoblast proliferation; Insulin signaling; Mevalonate; Muscle regeneration; Myotoxicity; Simvastatin

DOI:  https://doi.org/10.1016/j.bcp.2021.114649
J Cell Sci. 2021 Jun 24. pii: jcs.256388. [Epub ahead of print]

mRNA distribution in skeletal muscle is associated with mRNA size.

Helena Pinheiro, Mafalda Ramos Pimentel, Catarina Sequeira, Luís Manuel Oliveira, Anna Pezzarossa, William Roman, Edgar Rodrigues Gomes.

  Skeletal muscle myofibers are large and elongated cells with multiple and evenly distributed nuclei. Nuclear distribution suggests that each nucleus influences a specific compartment within the myofiber and implies a functional role for nuclear positioning. Compartmentalization of specific mRNAs and proteins has been reported at the neuromuscular and myotendinous junctions, but mRNA distribution in non-specialized regions of the myofibers remains largely unexplored. We report that the bulk of mRNAs is enriched around the nucleus of origin and that this perinuclear accumulation depends on recently transcribed mRNAs. Surprisingly, mRNAs encoding large proteins - giant mRNAs - are spread throughout the cell and do not exhibit perinuclear accumulation. Furthermore, by expressing exogenous transcripts with different sizes we found that size contributes to mRNA spreading independently of mRNA sequence. Both these mRNA distribution patterns depend on microtubules and are independent of nuclear dispersion, mRNA expression level and stability, and the characteristics of the encoded protein. Thus, we propose that mRNA distribution in non-specialized regions of skeletal muscle is size selective to ensure cellular compartmentalization and simultaneous long-range distribution of giant mRNAs.

Keywords:  compartmentalization; mRNA distribution; mRNA size; mRNA transport; multinucleation; skeletal muscle

DOI:  https://doi.org/10.1242/jcs.256388
J Gen Physiol. 2021 Jul 05. pii: e202112925. [Epub ahead of print]153(7):

Muscle ankyrin repeat protein 1 (MARP1) locks titin to the sarcomeric thin filament and is a passive force regulator.

Robbert J van der Pijl, Marloes van den Berg, Martijn van de Locht, Shengyi Shen, Sylvia J P Bogaards, Stefan Conijn, Paul Langlais, Pleuni E Hooijman, Siegfried Labeit, Leo M A Heunks, Henk Granzier, Coen A C Ottenheijm.

Muscle ankyrin repeat protein 1 (MARP1) is frequently up-regulated in stressed muscle, but its effect on skeletal muscle function is poorly understood. Here, we focused on its interaction with the titin-N2A element, found in titin's molecular spring region. We show that MARP1 binds to F-actin, and that this interaction is stronger when MARP1 forms a complex with titin-N2A. Mechanics and super-resolution microscopy revealed that MARP1 "locks" titin-N2A to the sarcomeric thin filament, causing increased extension of titin's elastic PEVK element and, importantly, increased passive force. In support of this mechanism, removal of thin filaments abolished the effect of MARP1 on passive force. The clinical relevance of this mechanism was established in diaphragm myofibers of mechanically ventilated rats and of critically ill patients. Thus, MARP1 regulates passive force by locking titin to the thin filament. We propose that in stressed muscle, this mechanism protects the sarcomere from mechanical damage.

DOI: https://doi.org/10.1085/jgp.202112925
Sci Rep. 2021 Jun 23. 11(1): 13197

Gelsolin and dCryAB act downstream of muscle identity genes and contribute to preventing muscle splitting and branching in Drosophila.

Benjamin Bertin, Yoan Renaud, Teresa Jagla, Guillaume Lavergne, Cristiana Dondi, Jean-Philippe Da Ponte, Guillaume Junion, Krzysztof Jagla.

A combinatorial code of identity transcription factors (iTFs) specifies the diversity of muscle types in Drosophila. We previously showed that two iTFs, Lms and Ap, play critical role in the identity of a subset of larval body wall muscles, the lateral transverse (LT) muscles. Intriguingly, a small portion of ap and lms mutants displays an increased number of LT muscles, a phenotype that recalls pathological split muscle fibers in human. However, genes acting downstream of Ap and Lms to prevent these aberrant muscle feature are not known. Here, we applied a cell type specific translational profiling (TRAP) to identify gene expression signatures underlying identity of muscle subsets including the LT muscles. We found that Gelsolin (Gel) and dCryAB, both encoding actin-interacting proteins, displayed LT muscle prevailing expression positively regulated by, the LT iTFs. Loss of dCryAB function resulted in LTs with irregular shape and occasional branched ends also observed in ap and lms mutant contexts. In contrast, enlarged and then split LTs with a greater number of myonuclei formed in Gel mutants while Gel gain of function resulted in unfused myoblasts, collectively indicating that Gel regulates LTs size and prevents splitting by limiting myoblast fusion. Thus, dCryAB and Gel act downstream of Lms and Ap and contribute to preventing LT muscle branching and splitting. Our findings offer first clues to still unknown mechanisms of pathological muscle splitting commonly detected in human dystrophic muscles and causing muscle weakness.

DOI: https://doi.org/10.1038/s41598-021-92506-3
Aging (Albany NY). 2021 Jun 24. 13

The consequences of a high-calorie diet background before calorie restriction on skeletal muscles in a mouse model.

Martin Maldonado, Jianying Chen, Yang Lujun, Huiqin Duan, Mazhar Ali Raja, Ting Qu, Tianhua Huang, Jiang Gu, Ying Zhong.

  The beneficial effects of calorie restriction (CR) are numerous. However, there is no scientific evidence about how a high-calorie diet (HCD) background influences the mechanisms underlying CR on skeletal muscles in an experimental mouse model. Herein we present empirical evidence showing significant interactions between HCD (4 months) and CR (3 months). Pectoralis major and quadriceps femoris vastus medialis, in the experimental and control groups, displayed metabolic and physiologic heterogeneity and remarkable plasticity, according to the dietary interventions. HCD-CR not only altered genetic activation patterns of satellite SC markers but also boosted the expression of myogenic regulatory factors and key activators of mitochondrial biogenesis, which in turn were also associated with metabolic fiber transition. Our data prompt us to theorize that the effects of CR may vary according to the physiologic, metabolic, and genetic peculiarities of the skeletal muscle described here and that INTM/IM lipid infiltration and tissue-specific fuel-energy status (demand/supply) both hold dependent-interacting roles with other key anti-aging mechanisms triggered by CR. Systematic integration of an HCD with CR appears to bring potential benefits for skeletal muscle function and energy metabolism. However, at this stage of our research, an optimal balance between the two dietary conditions, where anti-aging effects can be accomplished, is under intensive investigation in combination with other tissues and organs at different levels of organization within the organ system.

Keywords:  adiponectin; calorie-intervention; mitochondria; sirtuins; skeletal muscle

DOI:  https://doi.org/10.18632/aging.203237
Radiology. 2021 Jun 22. 204500

Hyperpolarized 13C MR Spectroscopy Depicts in Vivo Effect of Exercise on Pyruvate Metabolism in Human Skeletal Muscle.

Jae Mo Park, Crystal E Harrison, Junjie Ma, Jun Chen, James Ratnakar, Zungho Zun, Jeff Liticker, Galen D Reed, Avneesh Chhabra, Ronald G Haller, Thomas Jue, Craig R Malloy.

Background Pyruvate dehydrogenase (PDH) and lactate dehydrogenase are essential for adenosine triphosphate production in skeletal muscle. At the onset of exercise, oxidation of glucose and glycogen is quickly enabled by dephosphorylation of PDH. However, direct measurement of PDH flux in exercising human muscle is daunting, and the net effect of covalent modification and other control mechanisms on PDH flux has not been assessed. Purpose To demonstrate the feasibility of assessing PDH activation and changes in pyruvate metabolism in human skeletal muscle after the onset of exercise using carbon 13 (13C) MRI with hyperpolarized (HP) [1-13C]-pyruvate. Materials and Methods For this prospective study, sedentary adults in good general health (mean age, 42 years ± 18 [standard deviation]; six men) were recruited from August 2019 to September 2020. Subgroups of the participants were injected with HP [1-13C]-pyruvate at resting, during plantar flexion exercise, or 5 minutes after exercise during recovery. In parallel, hydrogen 1 arterial spin labeling MRI was performed to estimate muscle tissue perfusion. An unpaired t test was used for comparing 13C data among the states. Results At rest, HP [1-13C]-lactate and [1-13C]-alanine were detected in calf muscle, but [13C]-bicarbonate was negligible. During moderate flexion-extension exercise, total HP 13C signals (tC) increased 2.8-fold because of increased muscle perfusion (P = .005), and HP [1-13C]-lactate-to-tC ratio increased 1.7-fold (P = .04). HP [13C]-bicarbonate-to-tC ratio increased 8.4-fold (P = .002) and returned to the resting level 5 minutes after exercise, whereas the lactate-to-tC ratio continued to increase to 2.3-fold as compared with resting (P = .008). Conclusion Lactate and bicarbonate production from hyperpolarized (HP) [1-carbon 13 {13C}]-pyruvate in skeletal muscle rapidly reflected the onset and the termination of exercise. These results demonstrate the feasibility of imaging skeletal muscle metabolism using HP [1-13C]-pyruvate MRI and the sensitivity of in vivo pyruvate metabolism to exercise states. © RSNA, 2021 Online supplemental material is available for this article.

DOI: https://doi.org/10.1148/radiol.2021204500
EMBO Mol Med. 2021 Jun 21. e13695

Pathomechanisms and biomarkers in facioscapulohumeral muscular dystrophy: roles of DUX4 and PAX7.

Christopher R S Banerji, Peter S Zammit.

  Facioscapulohumeral muscular dystrophy (FSHD) is characterised by progressive skeletal muscle weakness and wasting. FSHD is linked to epigenetic derepression of the subtelomeric D4Z4 macrosatellite at chromosome 4q35. Epigenetic derepression permits the distal-most D4Z4 unit to transcribe DUX4, with transcripts stabilised by splicing to a poly(A) signal on permissive 4qA haplotypes. The pioneer transcription factor DUX4 activates target genes that are proposed to drive FSHD pathology. While this toxic gain-of-function model is a satisfying "bottom-up" genotype-to-phenotype link, DUX4 is rarely detectable in muscle and DUX4 target gene expression is inconsistent in patients. A reliable biomarker for FSHD is suppression of a target gene score of PAX7, a master regulator of myogenesis. However, it is unclear how this "top-down" finding links to genomic changes that characterise FSHD and to DUX4. Here, we explore the roles and interactions of DUX4 and PAX7 in FSHD pathology and how the relationship between these two transcription factors deepens understanding via the immune system and muscle regeneration. Considering how FSHD pathomechanisms are represented by "DUX4opathy" models has implications for developing therapies and current clinical trials.

Keywords:  DUX4; PAX7; biomarker; facioscapulohumeral muscular dystrophy (FSHD); pathology

DOI:  https://doi.org/10.15252/emmm.202013695
Nat Commun. 2021 06 22. 12(1): 3852

Fibroblast fusion to the muscle fiber regulates myotendinous junction formation.

Wesal Yaseen, Ortal Kraft-Sheleg, Shelly Zaffryar-Eilot, Shay Melamed, Chengyi Sun, Douglas P Millay, Peleg Hasson.

Vertebrate muscles and tendons are derived from distinct embryonic origins yet they must interact in order to facilitate muscle contraction and body movements. How robust muscle tendon junctions (MTJs) form to be able to withstand contraction forces is still not understood. Using techniques at a single cell resolution we reexamine the classical view of distinct identities for the tissues composing the musculoskeletal system. We identify fibroblasts that have switched on a myogenic program and demonstrate these dual identity cells fuse into the developing muscle fibers along the MTJs facilitating the introduction of fibroblast-specific transcripts into the elongating myofibers. We suggest this mechanism resulting in a hybrid muscle fiber, primarily along the fiber tips, enables a smooth transition from muscle fiber characteristics towards tendon features essential for forming robust MTJs. We propose that dual characteristics of junctional cells could be a common mechanism for generating stable interactions between tissues throughout the musculoskeletal system.

DOI: https://doi.org/10.1038/s41467-021-24159-9
Endocrinol Diabetes Nutr (Engl Ed). 2021 Mar;pii: S2530-0180(21)00036-6. [Epub ahead of print]68(3): 159-169

Effects of physical exercise in sarcopenia. A systematic review.

David E Barajas-Galindo, Elena González Arnáiz, Pablo Ferrero Vicente, María D Ballesteros-Pomar.

  From the third decade of life and due to multiple causes, muscle mass and strength are gradually lost, which affects the function of the musculoskeletal system. This combined loss of muscle mass and strength with aging is called sarcopenia, and is associated with greater morbidity and mortality in the elderly. Early treatment is therefore essential, and physical exercise is the therapeutic approach that has given the best results. This literature review intended to analyze the effect of physical exercise, excluding the role that other treatments proposed, including the nutritional approach, could play in the treatment of sarcopenia, refers to 12 articles. In studies including high intensity strength exercises in isolation, either alone or combined with aerobic exercise, improvements were seen in muscle mass, muscle strength, and functional test times. There is also a significant increase in fat-free mass in individuals who exercised more frequently (more than two sessions per week). Current evidence shows that strength-resistance training and its combination in multimodal programs with aerobic exercise show significantly beneficial effects on anthropometric and muscle function parameters. Programs of prescribed exercises including strength exercises adequate to the characteristics should therefore be adapted to the characteristics of each individual and replace the usual practice of prescribing aerobic exercises (walking) only.

Keywords:  Deporte en anciano; Ejercicio en anciano; Elderly; Envejecimiento activo; Older adults; Physical exercise in elderly; Sarcopenia

DOI:  https://doi.org/10.1016/j.endien.2020.02.007
Sci Adv. 2021 Jun;pii: eabh3693. [Epub ahead of print]7(26):

Anti-inflammatory nanoparticles significantly improve muscle function in a murine model of advanced muscular dystrophy.

Theresa M Raimondo, David J Mooney.

Chronic inflammation contributes to the pathogenesis of all muscular dystrophies. Inflammatory T cells damage muscle, while regulatory T cells (Tregs) promote regeneration. We hypothesized that providing anti-inflammatory cytokines in dystrophic muscle would promote proregenerative immune phenotypes and improve function. Primary T cells from dystrophic (mdx) mice responded appropriately to inflammatory or suppressive cytokines. Subsequently, interleukin-4 (IL-4)- or IL-10-conjugated gold nanoparticles (PA4, PA10) were injected into chronically injured, aged, mdx muscle. PA4 and PA10 increased T cell recruitment, with PA4 doubling CD4+/CD8- T cells versus controls. Further, 50% of CD4+/CD8- T cells were immunosuppressive Tregs following PA4, versus 20% in controls. Concomitant with Treg recruitment, muscles exhibited increased fiber area and fourfold increases in contraction force and velocity versus controls. The ability of PA4 to shift immune responses, and improve dystrophic muscle function, suggests that immunomodulatory treatment may benefit many genetically diverse muscular dystrophies, all of which share inflammatory pathology.

DOI: https://doi.org/10.1126/sciadv.abh3693
Nat Commun. 2021 06 22. 12(1): 3851

Unexpected contribution of fibroblasts to muscle lineage as a mechanism for limb muscle patterning.

Joana Esteves de Lima, Cédrine Blavet, Marie-Ange Bonnin, Estelle Hirsinger, Glenda Comai, Laurent Yvernogeau, Marie-Claire Delfini, Léa Bellenger, Sébastien Mella, Sonya Nassari, Catherine Robin, Ronen Schweitzer, Claire Fournier-Thibault, Thierry Jaffredo, Shahragim Tajbakhsh, Frédéric Relaix, Delphine Duprez.

Positional information driving limb muscle patterning is contained in connective tissue fibroblasts but not in myogenic cells. Limb muscles originate from somites, while connective tissues originate from lateral plate mesoderm. With cell and genetic lineage tracing we challenge this model and identify an unexpected contribution of lateral plate-derived fibroblasts to the myogenic lineage, preferentially at the myotendinous junction. Analysis of single-cell RNA-sequencing data from whole limbs at successive developmental stages identifies a population displaying a dual muscle and connective tissue signature. BMP signalling is active in this dual population and at the tendon/muscle interface. In vivo and in vitro gain- and loss-of-function experiments show that BMP signalling regulates a fibroblast-to-myoblast conversion. These results suggest a scenario in which BMP signalling converts a subset of lateral plate mesoderm-derived cells to a myogenic fate in order to create a boundary of fibroblast-derived myonuclei at the myotendinous junction that controls limb muscle patterning.

DOI: https://doi.org/10.1038/s41467-021-24157-x
Sci Rep. 2021 Jun 21. 11(1): 12993

Plasma lipidomic analysis shows a disease progression signature in mdx mice.

Roula Tsonaka, Alexandre Seyer, Annemieke Aartsma-Rus, Pietro Spitali.

Duchenne muscular dystrophy (DMD) is a rare genetic disorder affecting paediatric patients. The disease course is characterized by loss of muscle mass, which is rapidly substituted by fibrotic and adipose tissue. Clinical and preclinical models have clarified the processes leading to muscle damage and myofiber degeneration. Analysis of the fat component is however emerging as more evidence shows how muscle fat fraction is associated with patient performance and prognosis. In this article we aimed to study whether alterations exist in the composition of lipids in plasma samples obtained from mouse models. Analysis of plasma samples was performed in 4 mouse models of DMD and wild-type mice by LC-MS. Longitudinal samplings of individual mice covering an observational period of 7 months were obtained to cover the different phases of the disease. We report clear elevation of glycerolipids and glycerophospholipids families in dystrophic mice compared to healthy mice. Triacylglycerols were the strongest contributors to the signatures in mice. Annotation of individual lipids confirmed the elevation of lipids belonging to these families as strongest discriminants between healthy and dystrophic mice. A few sphingolipids (such as ganglioside GM2, sphingomyelin and ceramide), sterol lipids (such as cholesteryl oleate and cholesteryl arachidonate) and a fatty acyl (stearic acid) were also found to be affected in dystrophic mice. Analysis of serum and plasma samples show how several lipids are affected in dystrophic mice affected by muscular dystrophy. This study sets the basis to further investigations to understand how the lipid signature relates to the disease biology and muscle performance.

DOI: https://doi.org/10.1038/s41598-021-92406-6
Methods Mol Biol. 2021 ;2348 71-90

Single-Cell RNAseq Analysis of lncRNAs.

Stefano Cagnin, Enrico Alessio, Raphael Severino Bonadio, Gabriele Sales.

  Mammalian genomes are pervasively transcribed and a small fraction of RNAs produced codify for proteins. The importance of noncoding RNAs for the maintenance of cell functions is well known (e.g., rRNAs, tRNAs), but only recently it was first demonstrated the involvement of microRNAs (miRNAs) in posttranscriptional regulation and then the activity of long noncoding RNAs (lncRNAs) in the regulation of miRNAs, DNA structure and protein function. LncRNAs have an expression more cell specific than other RNAs and basing on their subcellular localization exert different functions. In this book chapter we consider different protocols to evaluate the expression of lncRNAs at the single cell level using genome-wide approaches. We considered the skeletal muscle as example because the most abundant tissue in mammals involved in the regulation of metabolism and body movement. We firstly described how to isolate the smallest complete contractile system responsible for muscle metabolic and contractile traits (myofibers). We considered how to separate long and short RNAs to allow the sequencing of the full-length transcript using the SMART technique for the retrotranscription. Because of myofibers are multinucleated cells and because of it is better to perform single cell sequencing on fresh tissues we described the single-nucleus sequencing that can be applied to frozen tissues. The chapter concludes with a description of bioinformatics approaches to evaluate differential expression from single-cell or single-nucleus RNA sequencing.

Keywords:  Bioinformatic; Long noncoding RNA; Myofiber; Single-nucleus RNA sequencing

DOI:  https://doi.org/10.1007/978-1-0716-1581-2_5
Biomed Pharmacother. 2021 Jun 17. pii: S0753-3322(21)00602-8. [Epub ahead of print]141 111820

Functional roles of non-coding RNAs in atrophy.

Soudeh Ghafouri-Fard, Atefe Abak, Shiva Khademi, Hamed Shoorei, Zahra Bahroudi, Mohammad Taheri, Nader Akbari Dilmaghani.

  Atrophy is defined as a reduction in cell, organ, or tissue size after reaching their normal mature sizes because of loss of organelles, cytoplasmic compartments, and proteins. This process is also involved in the pathogenesis of human disorders. Inadequate nourishment, poor circulation, inadequate hormonal support, defects in nerve supply of the tissue, disproportionate induction of apoptosis in the tissue, and absence of exercise are some underlying causes of atrophy. Recently, several non-coding RNAs (ncRNAs) have been identified that regulate atrophy, thus participating in the pathobiology of related disorders such as neurodegenerative/ neuromuscular diseases, age-related muscle atrophy, and cardiac tissue atrophy. In the current review, we have focused on two classes of ncRNAs namely long ncRNAs (lncRNAs) and microRNAs (miRNAs) to unravel their participation in atrophy-associated disorders.

Keywords:  Atrophy; Biomarkers; Expression; LncRNA; MiRNA

DOI:  https://doi.org/10.1016/j.biopha.2021.111820
Mol Cell Endocrinol. 2021 Jun 22. pii: S0303-7207(21)00213-6. [Epub ahead of print] 111369

Development of metabolic dysfunction in mice lacking chemerin.

Penghua Fang, Long Han, Mei Yu, Shiyu Han, Mengyuan Wan, Yujie Huang, Wancheng Guo, Qingbo Wei, Wenbing Shang, Wen Min.

  Chemerin, an adipocyte-secreted adipokine, is hypothesized to participate in energy homeostasis and glucoregulation. However, the physiologic effect of endogenous chemerin on glucose metabolism is unclear. The present studies tested the hypotheses that chemerin deficiency alters whole-body glucose homeostasis following switches to high-fat diet. Adult, male chemerin knockout and C57BL/6J control wild type mice were studied. During the following 4 weeks, chow- or high-fat diet maintained chemerin knockout mice showed elevated fasting glucose levels and glucose intolerance as well as insulin intolerance. Chemerin deficiency impaired adaptation to glucose and insulin challenge, leading to increased glucose levels. Moreover, the mRNA and protein levels of GLUT4 and PGC-1α expression in both skeletal muscle and adipose tissue were significantly decreased in chemerin knockout mice relative to the wild type, respectively. Taken together, the results support the hypotheses that chemerin helps adapt glucose metabolism to changes in dietary fat and modulates glucose consumption in mice by activation of PGC-1α/GLUT4 axis. Chemerin may play a significant role in elevation of glucose uptake and insulin sensitivity to promote glucose clearance.

Keywords:  Chemerin; GLUT4; PGC-1α; glucose metabolism

DOI:  https://doi.org/10.1016/j.mce.2021.111369
J Physiol. 2021 Jun 24.

Muscle clock: how does muscle physiology change in 24 hours in response to exercise?

Ana Paula Azevêdo Macêdo, Adelino Sanchez Ramos da Silva, José Rodrigo Pauli.

DOI: https://doi.org/10.1113/JP282005
J Biomech. 2021 Jun 07. pii: S0021-9290(21)00321-3. [Epub ahead of print]124 110540

A MATLAB toolbox for muscle diffusion-tensor MRI tractography.

Bruce M Damon, Zhaohua Ding, Melissa T Hooijmans, Adam W Anderson, Xingyu Zhou, Crystal L Coolbaugh, Mark K George, Bennett A Landman.

  Diffusion-tensor MRI fiber tractography has been used to reconstruct skeletal muscle architecture, but remains a specialized technique using custom-written data processing routines. In this work, we describe the public release of a software toolbox having the following design objectives: accomplish the pre-processing tasks of file input, image registration, denoising, and diffusion-tensor calculation; allow muscle-specific methods for defining seed points; make fiber-tract architectural measurements referenced to tendinous structures; visualize fiber tracts and other muscle structures of interest; analyze the goodness of outcomes; and provide a programming structure that allows the addition of new capabilities in future versions. The proper function of the code was verified using simulated datasets. The toolbox capabilities for characterizing human muscle structure in vivo were demonstrated in a case study. These capabilities included measurements of muscle morphology; contractile and non-contractile tissue volumes; fiber-tract length, pennation angle, curvature; and the physiological cross-sectional area,. The free public release of this software is a first step in creating of a community of users who use these tools in studies of muscle physiology and biomechanics. Users may further contribute to code development. Along with simulated and actual datasets for benchmarking, these tools will further create mechanisms for enhancing scientific rigor and developing and validating new code features. Planned future developments include additional options for image pre-processing, development of a graphical user interface, analysis of architectural patterns during muscle contraction, and integration of functional imaging data.

Keywords:  Magnetic resonance imaging; Open research; Skeletal muscle; Software

DOI:  https://doi.org/10.1016/j.jbiomech.2021.110540
BMC Genomics. 2021 Jun 24. 22(1): 469

Morphological changes and functional circRNAs screening of rabbit skeletal muscle development.

Qi Zheng, Cuiyun Zhu, Jing Jing, Yinghui Ling, Shuaiqi Qin, Jiao Wang, Lisha Zha, Ya Liu, Fugui Fang.

   BACKGROUND: The temporal expression pattern of circular RNAs (circRNAs) across developmental stages is essential for skeletal muscle growth and functional analysis. However, there are few analyses on the potential functions of circRNAs in rabbit skeletal muscle development.
RESULTS: Initially, the paraffin sections showed extremely significant differences in the diameter, number, area and density of skeletal muscle fibers of the fetus, child, adult rabbit hind legs (P < 0.01). Then, RNA-seq libraries of these three stages were constructed. A total of 481 differentially expressed circRNAs (DE-circRNAs) and 5,658 differentially expressed genes (DEGs) were identified. Subsequently, DE-circRNAs, whose host genes were DEGs or non-DEGs, were analyzed by GO respectively. In the fetus vs. child group, up-regulated DE-circRNAs (whose host genes were DEGs) were related to muscle fiber structure, and down-regulated ones were related to mitosis. The up-regulated DE-circRNAs (whose host genes were non-DEGs) were involved in enzyme activity, methylation and glycosylation, and the down-regulated ones were involved in mitosis and catabolism. In the fetus vs. adult group, the up-regulated DE-circRNAs (whose host genes were DEGs) were related to skeletal muscle basic structure, and the down-regulated ones were also associated with cell proliferation. But the up-regulated DE-circRNAs (whose host genes were non-DEGs) were connected with regulation of histone ubiquitination, chromatin and organelles. The down-regulated DE-circRNAs were connected with the catabolism processes. In addition, novel_circ_0022663 and novel_circ_0005489, which might have coding potential, and novel_circ_0004210 and novel_circ_0001669, which might have miRNA sponge capability, were screened out.
CONCLUSIONS: In this study, hind leg muscles of fetus, child and adult rabbits were collected for paraffin section and RNA-seq to observe the structural changes of skeletal muscle and obtain circRNA expression profiles at different stages. These data provided a catalog of circRNAs related to muscle development in New Zealand rabbits, allowing us to better understand the functional transitions in mammalian muscle development.

Keywords:  CircRNA; Morphology; RNA-seq; Rabbit; Skeletal muscle

DOI:  https://doi.org/10.1186/s12864-021-07706-y
J Appl Physiol (1985). 2021 06 24.

Neural adaptations to long-term resistance training: evidence for the confounding effect of muscle size on the interpretation of surface electromyography.

Jakob Škarabot, Thomas Grant Balshaw, Sumiaki Maeo, Garry J Massey, Marcel Bahia Lanza, Thomas M Maden-Wilkinson, Jonathan P Folland.

  This study compared elbow flexor (EF; Experiment 1) and knee extensor (KE; Experiment 2) maximal compound action potential (Mmax) amplitude between long-term resistance trained (LTRT; n=15 and n=14, 6±3 and 4±1 years of training) and untrained (UT; n=14 and n=49) men; and examined the effect of normalising electromyography (EMG) during maximal voluntary torque (MVT) production to Mmax amplitude on differences between LTRT and UT. EMG was recorded from multiple sites and muscles of EF and KE, Mmax was evoked with percutaneous nerve stimulation, and muscle size was assessed with ultrasonography (thickness, EF) and magnetic resonance imaging (cross-sectional area, KE). Muscle-electrode distance (MED) was measured to account for the effect of adipose tissue on EMG and Mmax. LTRT displayed greater MVT (+66-71%, p<0.001), muscle size (+54-56%, p<0.001), and Mmax amplitudes (+29-60%, p≤0.010) even when corrected for MED (p≤0.045). Mmax was associated with the size of both muscle groups (r≥0.466, p≤0.011). Compared to UT, LTRT had higher absolute voluntary EMG amplitude for the KE (p<0.001), but not the EF (p=0.195), and these differences/similarities were maintained after correction for MED; however, Mmax normalisation resulted in no differences between LTRT and UT for any muscle and/or muscle group (p≥0.652). The positive association between Mmax and muscle size, and no differences when accounting for peripheral electrophysiological properties (EMG/Mmax), indicates the greater absolute voluntary EMG amplitude of LTRT might be confounded by muscle morphology, rather than provide a discrete measure of central neural activity. This study therefore suggests limited agonist neural adaptation after LTRT.

Keywords:  M-wave; Muscle excitability; Sarcolemmal excitability; Strength training; Surface electromyography

DOI:  https://doi.org/10.1152/japplphysiol.00094.2021