Skeletal Muscle Disuse Atrophy Research Articles

Nutritional intervention to enhance recovery after arthroscopic knee surgery in adults: a randomized controlled pilot trial

BackgroundEssential amino acid (EAA) and omega-3 fatty acid ingestion independently attenuate leg skeletal muscle disuse atrophy in uninjured persons. However, no data exist regarding the effectiveness of combined EAA and omega-3 fatty acid ingestion to mitigate skeletal muscle disuse atrophy in response to anterior cruciate ligament reconstruction (ACLR) surgery. This pilot trial will explore the feasibility of recruitment and retention of ACLR outpatients from a single center across 18 months to consume either a combination of omega-3 fatty acids and EAAs, or a placebo control, for 4 weeks before and 2 weeks after surgery.MethodsThirty adult (≥ 18 years old) ACLR outpatients will be recruited for this single center, double-blind, two-arm randomized controlled feasibility pilot trial. Participants will consume either 5 g⋅day−1 of omega-3 fatty acids (fish oil) and 40 g⋅day−1 of EAAs or 5 g⋅day−1 of a control fatty acid mixture (safflower oil) and 40 g⋅day−1 of non-essential amino acids (NEAAs). Fatty acid supplements will be consumed 4 weeks before and for 2 weeks after ACLR surgery, whereas the EAAs and NEAAs will be consumed 1 week before and for 2 weeks after ACLR surgery. The primary outcomes are feasibility of recruitment and retention, with the goal to recruit 30 outpatients across 18 months and retain 22 participants upon completion of the study protocol following 12 weeks of data collection. These results will be reported using descriptive statistics, along with reasons and timepoints for study dropout. Secondary exploratory outcomes will be reported using inferential statistics for purposes of hypothesis generation and elucidation of mechanistic targets for future work; no inferences to clinical efficacy will be made. These outcomes include integrated rates of skeletal muscle protein synthesis, skeletal muscle protein content and expression of translation factors, skeletal muscle and erythrocyte phospholipid composition, and measures of skeletal muscle mass, strength, and power.ImpactThis work will set the foundation for a future randomized controlled trial powered to detect an effect of EAA + omega-3 fatty acid intake on skeletal muscle size or function in response to ACLR surgery.Trial registrationClinicalTrials.gov, NCT06233825. Registered 31 January 2024. https://clinicaltrials.gov/study/NCT06233825?term=NCT06233825&rank=1

O-GlcNAcylation of SERCA protects skeletal muscle in hibernating Spermophilus dauricus from disuse atrophy

Long-term inactivity of skeletal muscle results in muscular disuse atrophy; however, hibernating animals do not experience muscular disuse atrophy during the hibernation period. The molecular mechanism underlining the anti-atrophy effect in these animals is unclear. O-linked N acetyl-β-D-glucosaminylation (O-GlcNAcylation) and its effect on cell signaling pathways are important mechanisms underlying muscular disuse atrophy; thus, in this study, we investigated O-GlcNAcylation changes during hibernation in Spermophilus dauricus to explore the role of O-GlcNAcylation in the muscle disuse atrophy resistance of hibernating animals. The results showed that during hibernation, the muscle fiber cross-sectional area and ratio of muscle fiber did not change, and the morphological structure of the muscle remained intact, with normal contractile function. The level of O-GlcNAcylation decreased during hibernation, but quickly returned to normal in the periodic arousal stage. The O-GlcNAcylation level of sarcoplasmic/endoplasmic reticulum calcium ATPase 1 (SERCA1) decreased, whereas its activity increased. The decrease in O-GlcNAcylation of SERCA could result in the decreased binding of phospholamban to SERCA1, thus decreasing its inhibition to SERCA1 activity. This in turn can inhibit muscle cell calcium overload, maintain muscle cell calcium homeostasis, and stabilize the calpain proteolytic pathway, ultimately inhibiting skeletal muscle atrophy. Our results demonstrate that periodic arousal along with returning O-GlcNAcylation level to normal are important mechanisms in preventing disuse atrophy of skeletal muscle during hibernation.

Proteomic Identification of Seasonally Expressed Proteins Contributing to Heart Function and the Avoidance of Skeletal Muscle Disuse Atrophy in a Hibernating Mammal.

Hibernation in the thirteen-lined ground squirrel (Ictidomys tridecemlineatus) takes place over 4-6 months and is characterized by multiday bouts of hypothermic torpor (5-7 °C core body temperature) that are regularly interrupted every 1-2 weeks by brief (12-24 h) normothermic active periods called interbout arousals. Our goal was to gain insight into the molecular mechanisms that underlie the hibernator's ability to preserve heart function and avoid the deleterious effects of skeletal muscle disuse atrophy over prolonged periods of inactivity, starvation, and near-freezing body temperatures. To achieve this goal, we performed organelle enrichment of heart and skeletal muscle at five seasonal time points followed by LC-MS-based label-free quantitative proteomics. In both organs, we saw an increase in the levels of many proteins as ground squirrels transition from an active state to a prehibernation state in the fall. Interestingly, seasonal abundance patterns identified DHRS7C, SRL, TRIM72, RTN2, and MPZ as potential protein candidates for mitigating disuse atrophy in skeletal muscle, and ex vivo contractile mechanics analysis revealed no deleterious effects in the ground squirrel's muscles despite prolonged sedentary activity. Overall, an increased understanding of protein abundance in hibernators may enable novel therapeutic strategies to treat muscle disuse atrophy and heart disease in humans.

Plasticity changes in neuromuscular junction morphology and related regulatory proteins in the hibernating ground squirrel.

Skeletal muscle disuse atrophy can cause degenerative changes in neuromuscular junction morphology. Although Daurian ground squirrels (Spermophilus dauricus) are a natural anti-disuse animal model for studying muscle atrophy during hibernation, little is known about the morphological and regulatory mechanisms of their neuromuscular junctions. Here, we found that morphological indices of the soleus muscle were significantly lower during hibernation (torpor and interbout arousal) compared to pre-hibernation but recovered during post-hibernation. In the extensor digitorum longus muscle, neuromuscular junction morphology did not change significantly during hibernation. Agrin-Lrp4-MuSK is a key pathway for the formation and maintenance of the neuromuscular junction. Our results showed that low-density lipoprotein receptor-associated protein 4 (Lrp4) expression in the soleus (slow muscle) decreased by 46.2% in the interbout arousal group compared with the pre-hibernation group (p = 0.019), with recovery in the post-hibernation group. Compared to the pre-hibernation group, agrin expression in the extensor digitorum longus (fast muscle) increased by 67.0% in the interbout arousal group (p = 0.016). In conclusion, periodic up-regulation in agrin expression during interbout arousal may be involved in the maintenance of neuromuscular junction morphology in the extensor digitorum longus muscle during hibernation. The degenerative changes in neuromuscular junction morphology and the periodic decrease in Lrp4 protein expression in the soleus during hibernation, these changes recovered to the pre-hibernation levels in the post-hibernation group, also exhibiting significant plasticity. This plasticity may be an important mechanism for resisting disuse atrophy in hibernating animals.

New Findings: Hindlimb Unloading Causes Nucleocytoplasmic Ca2+ Overload and DNA Damage in Skeletal Muscle.

Disuse atrophy of skeletal muscle is associated with a severe imbalance in cellular Ca2+ homeostasis and marked increase in nuclear apoptosis. Nuclear Ca2+ is involved in the regulation of cellular Ca2+ homeostasis. However, it remains unclear whether nuclear Ca2+ levels change under skeletal muscle disuse conditions, and whether changes in nuclear Ca2+ levels are associated with nuclear apoptosis. In this study, changes in Ca2+ levels, Ca2+ transporters, and regulatory factors in the nucleus of hindlimb unloaded rat soleus muscle were examined to investigate the effects of disuse on nuclear Ca2+ homeostasis and apoptosis. Results showed that, after hindlimb unloading, the nuclear envelope Ca2+ levels ([Ca2+]NE) and nucleocytoplasmic Ca2+ levels ([Ca2+]NC) increased by 78% (p < 0.01) and 106% (p < 0.01), respectively. The levels of Ca2+-ATPase type 2 (Ca2+-ATPase2), Ryanodine receptor 1 (RyR1), Inositol 1,4,5-tetrakisphosphate receptor 1 (IP3R1), Cyclic ADP ribose hydrolase (CD38) and Inositol 1,4,5-tetrakisphosphate (IP3) increased by 470% (p < 0.001), 94% (p < 0.05), 170% (p < 0.001), 640% (p < 0.001) and 12% (p < 0.05), respectively, and the levels of Na+/Ca2+ exchanger 3 (NCX3), Ca2+/calmodulin dependent protein kinase II (CaMK II) and Protein kinase A (PKA) decreased by 54% (p < 0.001), 33% (p < 0.05) and 5% (p > 0.05), respectively. In addition, DNase X is mainly localized in the myonucleus and its activity is elevated after hindlimb unloading. Overall, our results suggest that enhanced Ca2+ uptake from cytoplasm is involved in the increase in [Ca2+]NE after hindlimb unloading. Moreover, the increase in [Ca2+]NC is attributed to increased Ca2+ release into nucleocytoplasm and weakened Ca2+ uptake from nucleocytoplasm. DNase X is activated due to elevated [Ca2+]NC, leading to DNA fragmentation in myonucleus, ultimately initiating myonuclear apoptosis. Nucleocytoplasmic Ca2+ overload may contribute to the increased incidence of myonuclear apoptosis in disused skeletal muscle.

Cinnamtannin A2, (-)-epicatechin tetramer, attenuates skeletal muscle wasting in disuse atrophy model mice induced by hindlimb suspension.

The impact of repeated administration of cinntamtannin A2 (A2, 25 μg/kg) on skeletal muscle disuse atrophy model mice induced by hindlimb suspension for 14 days was examined. In soleus, weight loss and a reduction in the average myofibre size with shifting to the smaller side of the peak were observed in the suspension-vehicle group, but A2 reduced these changes. Average myofibre size significantly increased in ground-A2 compared to ground-vehicle. A marked increase in the dephosphorylation of forkhead box O (FoxO) 3a by the suspension was reduced by A2. The phosphorylation of protein kinase B (Akt) and eukaryotic translation initiation factor 4E-binding protein (4EBP)-1 were significantly increased by the treatment of A2. In addition, a single dose of A2 increased dramatically in the 24-h excretion of catecholamines in urine. These results suggest that A2 administration results in sympathetic nerve activation and promotes hypertrophy while inhibiting the progress of disuse muscle atrophy.

Construction and Analysis of Disuse Atrophy Model of the Gastrocnemius Muscle in Chicken.

Disuse muscle atrophy is identified as the physiological, biochemical, morphological, and functional changes during restricted movement, immobilization, or weightlessness. Although its internal mechanism has been extensively studied in mammals and was thought to be mainly related to oxidative stress, it was unclear whether it behaved consistently in non-mammals such as chickens. In this study, we tried to construct a disuse atrophy model of the gastrocnemius muscle in chickens by limb immobilization, and collected the gastrocnemius muscles of the fixed group and the control group for RNA sequencing. Through analysis of muscle loss, HE staining, immunohistochemistry, and oxidative stress level, we found that limb immobilization could lead to loss of muscle mass, decrease in muscle fiber diameter, decrease in the proportion of slow muscle fibers, and increase in the proportion of fast muscle fibers, and also cause elevated levels of oxidative stress. In addition, a total of 565 different expression genes (DEGs) were obtained by RNA sequencing, which was significantly enriched in the biological processes such as cell proliferation and apoptosis, reactive oxygen species metabolism, and fast and slow muscle fiber transformation, and it showed that the FOXO signaling pathway, closely related to muscle atrophy, was activated. In brief, we initially confirmed that limb immobilization could induce disuse atrophy of skeletal muscle, and oxidative stress was involved in the process of disuse muscle atrophy.

Declines in muscle protein synthesis account for short-term muscle disuse atrophy in humans in the absence of increased muscle protein breakdown.

BackgroundWe determined the short‐term (i.e. 4 days) impacts of disuse atrophy in relation to muscle protein turnover [acute fasted‐fed muscle protein synthesis (MPS)/muscle protein breakdown (MPB) and integrated MPS/estimated MPB].MethodsHealthy men (N = 9, 22 ± 2 years, body mass index 24 ± 3 kg m−2) underwent 4 day unilateral leg immobilization. Vastus lateralis (VL) muscle thickness (MT) and extensor strength and thigh lean mass (TLM) were measured. Bilateral VL muscle biopsies were collected on Day 4 at t = −120, 0, 90, and 180 min to determine integrated MPS, estimated MPB, acute fasted‐fed MPS (l‐[ring‐13C6]‐phe), and acute fasted tracer decay rate representative of MPB (l‐[15N]‐phe and l‐[2H8]‐phe). Protein turnover cell signalling was measured by immunoblotting.ResultsImmobilization decreased TLM [pre: 7477 ± 1196 g, post: 7352 ± 1209 g (P < 0.01)], MT [pre: 2.67 ± 0.50 cm, post: 2.55 ± 0.51 cm (P < 0.05)], and strength [pre: 260 ± 43 N m, post: 229 ± 37 N m (P < 0.05)] with no change in control legs. Integrated MPS decreased in immob vs. control legs [control: 1.55 ± 0.21% day−1, immob: 1.29 ± 0.17% day−1 (P < 0.01)], while tracer decay rate (i.e. MPB) (control: 0.02 ± 0.006, immob: 0.015 ± 0.015) and fractional breakdown rate (FBR) remained unchanged [control: 1.44 ± 0.51% day−1, immob: 1.73 ± 0.35% day−1 (P = 0.21)]. Changes in MT correlated with those in MPS but not FBR. MPS increased in the control leg following feeding [fasted: 0.043 ± 0.012% h−1, fed: 0.065 ± 0.017% h−1 (P < 0.05)] but not in immob [fasted: 0.034 ± 0.014% h−1, fed: 0.049 ± 0.023% h−1 (P = 0.09)]. There were no changes in markers of MPB with immob (P > 0.05).ConclusionsHuman skeletal muscle disuse atrophy is driven by declines in MPS, not increases in MPB. Pro‐anabolic therapies to mitigate disuse atrophy would likely be more effective than therapies aimed at attenuating protein degradation.

Histological structure of skeletal muscle in disuse muscle atrophy and role of exercise

Disuse muscle atrophy is one of the most important research topics in fields of rehabilitation and clinical medicine. It refers to morphological and functional alteration of skeletal muscle under the states of hypokinesia, immobilization and weightlessness. This review analyzes a number of structural and functional changes in the skeletal muscle tissue in case of muscle disuse which appeared in loss in the muscle mass, reduction in the transverse diameter of most of muscle fibers, wide spaces between muscle fibers, increase collagen fiber deposition and appearance of apoptotic nuclei in muscle fibers besides sensorial dysfunction such as hypo mechanical responsiveness and diminished contractile function. In addition to the ultrastructural alteration, which include loss of sarcomere organization, indistinguishable A and I bands, irregular distorted Z-line with disruption of myofilaments and fused giant accumulated mitochondria. The review summarizes also the molecular mechanisms of skeletal muscle disuse atrophy which could be resulted from an alteration in the activity of oxidative stress enzymes and lead to increase in protein degradation and/or a reduction in protein synthesis. Moreover, the review discusses the efficiency of potential countermeasures of stretching exercise as a very powerful stimulant of muscle growth and muscle protein synthesis to counteract the structural and functional alterations in muscle tissue caused by disuse. The review showed that stretching exercise can increase the transverse sectional area of muscle fibers, produce a rise in the number of sarcomeres in series and prevent muscle atrophy. Passive stretching also completely prevented the decrease in myonuclear number induced by inactivity. Moreover, stretching accelerated the amelioration of the range of motion limitation and prevent the deposition of connective tissue resulted from disuse muscle atrophy.

Early mitochondrial degeneration in the development of disuse‐induced muscle atrophy

Skeletal muscle disuse and subsequent atrophy occurs with multiple pathological stimuli such as bedrest and spaceflight. It is known that during disuse glycolytic muscles appear largely protected while mitochondria‐dense oxidative muscles undergo rampant wasting, suggesting a possible mitochondrial component to this wasting. However, the role of mitochondrial degeneration in the development of disuse‐induced muscle atrophy is not well understood.PurposeTo examine the development of mitochondrial network degeneration using the fluorescent reporter pMitoTimer during the development of disuse‐induced muscle atrophy.MethodsAt 10 weeks of age, C57BL6/J mice underwent hindlimb suspension for 24, 48, 72, or 168 hours. Control mice were singly housed under normal cage conditions. To examine mitochondrial network quality the fluorescent reporter pMitoTimer was electroporated into the Flexor Digitorum Brevis (FDB) muscle two weeks prior to hindlimb suspension. pMitoTimer has been previously validated to assess mitochondrial quality using ratio of red:green fluorescence intensity with red fluorescence indicating degeneration. Additionally, detection of pure red puncta suggests completely degenerated mitochondria tagged for mitophagic degradation. At the appropriate timepoint, each animal was humanely euthanized and hindlimb muscles collected. The FDB was immediately fixed in 4% paraformaldehyde and imaged on Nikon TiS epifluorescent microscope with LED based light source and standardized image acquisition parameters. pMitoTimer fluorescence was quantified for ratio of red:green fluorescence intensity and pure red puncta using a previously developed Matlab program. Data were statistically analyzed by one‐way ANOVA followed by Tukey's posthoc test when significant effects were found and α was set at 0.05.ResultsAt 24 hours HS weights of soleus, plantaris and gastrocnemius muscles were not different from control. At 48 hours, however, weights of soleus, plantaris and gastrocnemius muscles were 11–14% lower than load‐bearing controls and at 168 hours weights of these muscles were ~16–39% lower than control with the greatest differences seen in the soleus. At 24 hours, pMitoTimer red:green ratio was ~37% greater compared to control and at 48 hours ~55% greater compared to control with no other statistical differences observed. Number of pMitoTimer pure red puncta was 5X greater in 48 hour HS mice compared to control and 24 hour with no other differences observed.ConclusionsDegeneration of the mitochondrial network measured by the fluorescent reporter pMitoTimer occurs prior to the onset of muscle atrophy following hindlimb suspension‐induced muscle disuse. These data may provide evidence for a novel mechanism for the instigation of muscle atrophy with disuse.Support or Funding InformationThis study was supported by National Institutes of Health R15AR069913.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Overview of FES-Assisted Cycling Approaches and Their Benefits on Functional Rehabilitation and Muscle Atrophy.

Central nervous system diseases include brain or spinal cord impairments and may result in movement disorders almost always manifested by paralyzed muscles with preserved innervations and therefore susceptible to be activated by electrical stimulation. Functional electrical stimulation (FES)-assisted cycling is an approach mainly used for rehabilitation purposes contributing, among other effects, to restore muscle trophism. FES-assisted cycling has also been adapted for mobile devices adding a leisure and recreational benefit to the physical training. In October 2016, our teams (Freewheels and EMA-trike) took part in FES-bike discipline at the Cybathlon competition, presenting technologies that allow pilots with spinal cord injury to use their paralyzed lower limb muscles to propel a tricycle. Among the many benefits observed and reported in our study cases for the pilots during preparation period, we achieved a muscle remodeling in response to FES-assisted cycling that is discussed in this chapter. Then, we have organized some sections to explore how FES-assisted cycling could contribute to functional rehabilitation by means of changes in the skeletal muscle disuse atrophy.

Disuse skeletal muscle atrophy in humans. Proteomic and molecular adaptations

Disuse skeletal muscle atrophy in humans. Proteomic and molecular adaptations

Partial or Complete Unloading of Skeletal Muscle Leads to Specific Alterations of Anabolic Signal Transduction

Consequences of disuse atrophy of skeletal muscle observed during spaceflight on astronaut health and performance are a focal point of space research. Decrements of both muscle mass and protein synthesis rates have been observed with exposure to varying muscle loading environments (1G > partial loading > 0G), and most of the reduced muscle mass can be attributed to diminished rates of synthesis. However, specific mechanisms behind unloading-dependent reductions of protein synthesis are not well defined. PURPOSE: To determine whether or not alterations of anabolic signal transduction was responsible for the changes previously observed in fractional synthesis rates with specific gravitational loading paradigms. METHODS: Female BALB/cByJ were normalized by bodyweight and assigned to normal cage ambulation (1G), partial weight bearing suspension titrated to approximately 33% bodyweight (G/3), partial weight bearing titrated to 16% bodyweight (G/6) and full unloading of hind limbs (0G) in specially designed cages. All mice were subjected to that loading environment for 21d prior to tissue harvest, and monitored daily. Immunoblotting of the gastrocnemius (n=23) was carried out to analyze alterations of anabolic signal transduction. Although numerous signaling intermediates were assessed, the focus of this abstract will be on ribosomal protein S6 kinase (p70-S6K). This important protein has served as a marker of protein synthesis signal transduction as well as the anabolic capacity in skeletal muscle. RESULTS: Regardless of loading paradigm, no differences were detected among groups for the activation of p70-S6K (as indicated by the phospho: total protein content). Total protein content, however, was ~27% lower than control in 0G and G/6 (P=0.008) with G/3 not being different from control (P>0.05). CONCLUSION: In combination with previous data (unpublished observations), ambulation at G/3 is sufficient to maintain anabolic signaling capacity when compared to G/6 or 0G, suggesting that a threshold level of stimulus is necessary to maintain anabolic capacity in muscle. These results may have important implications towards the development of strategies designed to counter the effects of partial/complete unloading on skeletal muscle based on how the anabolic capacity of muscle is affected.

Human Skeletal Muscle Disuse Atrophy: Effects on Muscle Protein Synthesis, Breakdown, and Insulin Resistance—A Qualitative Review

The ever increasing burden of an aging population and pandemic of metabolic syndrome worldwide demands further understanding of the modifiable risk factors in reducing disability and morbidity associated with these conditions. Disuse skeletal muscle atrophy (sometimes referred to as “simple” atrophy) and insulin resistance are “non-pathological” events resulting from sedentary behavior and periods of enforced immobilization e.g., due to fractures or elective orthopedic surgery. Yet, the processes and drivers regulating disuse atrophy and insulin resistance and the associated molecular events remain unclear—especially in humans. The aim of this review is to present current knowledge of relationships between muscle protein turnover, insulin resistance and muscle atrophy during disuse, principally in humans. Immobilization lowers fasted state muscle protein synthesis (MPS) and induces fed-state “anabolic resistance.” While a lack of dynamic measurements of muscle protein breakdown (MPB) precludes defining a definitive role for MPB in disuse atrophy, some proteolytic “marker” studies (e.g., MPB genes) suggest a potential early elevation. Immobilization also induces muscle insulin resistance (IR). Moreover, the trajectory of muscle atrophy appears to be accelerated in persistent IR states (e.g., Type II diabetes), suggesting IR may contribute to muscle disuse atrophy under these conditions. Nonetheless, the role of differences in insulin sensitivity across distinct muscle groups and its effects on rates of atrophy remains unclear. Multifaceted time-course studies into the collective role of insulin resistance and muscle protein turnover in the setting of disuse muscle atrophy, in humans, are needed to facilitate the development of appropriate countermeasures and efficacious rehabilitation protocols.

Angiotensin-(1-7) attenuates disuse skeletal muscle atrophy via the Mas receptor

ABSTRACTImmobilization is a form of disuse characterized by a loss of strength and muscle mass. Among the main features are decreased IGF-1/Akt signalling and increased ubiquitin-proteasome pathway signalling, which induce greater myosin heavy chain degradation. Activation of the classical renin-angiotensin system (RAS) causes deleterious effects in skeletal muscle, including muscle wasting. In contrast, angiotensin-(1-7) [Ang-(1-7)], a peptide of the non-classical RAS, produces beneficial effects in skeletal muscle. However, the role of Ang-(1-7) in skeletal muscle disuse atrophy and independent of classical RAS activation has not been evaluated. Therefore, we assessed the functions of Ang-(1-7) and the Mas receptor in disuse muscle atrophy in vivo using unilateral cast immobilization of the hind limb in male, 12-week-old wild-type (WT) and Mas-knockout (Mas KO) mice for 1 and 14 days. Additionally, we evaluated the participation of IGF-1/IGFR-1/Akt signalling and ubiquitin-proteasome pathway expression on the effects of Ang-(1-7) immobilization-induced muscle atrophy. Our results found that Ang-(1-7) prevented decreased muscle strength and reduced myofiber diameter, myosin heavy chain levels, and the induction of atrogin-1 and MuRF-1 expressions, all of which normally occur during immobilization. Analyses indicated that Ang-(1-7) increases IGF-1/IGFR-1/Akt pathway signalling through IGFR-1 and Akt phosphorylation, and the concomitant activation of two downstream targets of Akt, p70S6K and FoxO3. These anti-atrophic effects of Ang-(1-7) were not observed in Mas KO mice, indicating crucial participation of the Mas receptor. This report is the first to propose anti-atrophic effects of Ang-(1-7) via the Mas receptor and the participation of the IGF-1/IGFR-1/Akt/p70S6K/FoxO3 mechanism in disuse skeletal muscle atrophy.

Early Signs Of Extra Cellular Matrix Remodeling After Immobilization In Old And Young Human Individuals

Skeletal musle disuse has been shown to induce muscle atrophy rapidly in both young and old human individuals (1) as well as extracellular matrix (ECM) remodeling in young individuals (2). However, little is known about the effect of immobilization on ECM remodeling in older individuals. PURPOSE: To study the effect of aging on transcriptional regulatory signaling pathways involved in ECM remodeling with short-term immobilization. METHODS: Myofiber atrophy was induced by application of a knee-brace for a total period of 4 days in young (Y∼20 yrs, n=11) and aged (O∼70 yrs, n=11) individuals. Muscle biopsies of the vastus lateralis (VL) muscle were collected 1 week prior to immobilization and after 24h, 48h and 96h of immobilization. Expression levels of Col1A1, Col3A1, MMP2 and MMP9 mRNAs were determined using real-time RT-PCR and normalized to the Ribosomal Protein Large P0 (RPLP0) mRNA. RESULTS: In both age groups an increase in expression levels of Col1A1, Col3A1, MMP2 and MMP9 mRNA was observed after 24h. In contrast, expression levels of Col1A1 and Col3A1 mRNA were up regulated at 48h and 96h primarily in old individuals and an overall effect of age was observed. The expression level of MMP2 mRNA was not changed from pre at 48h and 96h. The expression level of MMP9 mRNA was up regulated in both age groups at all three time points. CONCLUSIONS: The present data demonstrates early (post 24h) signs of ECM remodeling in both young and old individuals. Moreover, the results point toward an age specific regulation of ECM in response to muscle disuse, with an up regulation of collagen I and III in old only after 48h and 96h of immobilisation. REFERENCES 1) Suetta C et al. Aging affects the transcriptional regulation of human skeletal muscle disuse atrophy. PLoS One. 2012;7(12) 2) Reich KA et al. Forty-eight hours of unloading and 24 h of reloading lead to changes in global gene expression patterns related to ubiquination and oxidative stress in humans. J Appl Physiol 109: 1404-1415, 2010.

PGC1-α over-expression prevents metabolic alterations and soleus muscle atrophy in hindlimb unloaded mice.

Prolonged skeletal muscle inactivity causes muscle fibre atrophy. Redox imbalance has been considered one of the major triggers of skeletal muscle disuse atrophy, but whether redox imbalance is actually the major cause or simply a consequence of muscle disuse remains of debate. Here we hypothesized that a metabolic stress mediated by PGC-1α down-regulation plays a major role in disuse atrophy. First we studied the adaptations of soleus to mice hindlimb unloading (HU) in the early phase of disuse (3 and 7 days of HU) with and without antioxidant treatment (trolox). HU caused a reduction in cross-sectional area, redox status alteration (NRF2, SOD1 and catalase up-regulation), and induction of the ubiquitin proteasome system (MuRF-1 and atrogin-1 mRNA up-regulation) and autophagy (Beclin1 and p62 mRNA up-regulation). Trolox completely prevented the induction of NRF2, SOD1 and catalase mRNAs, but not atrophy or induction of catabolic systems in unloaded muscles, suggesting that oxidative stress is not a major cause of disuse atrophy. HU mice showed a marked alteration of oxidative metabolism. PGC-1α and mitochondrial complexes were down-regulated and DRP1 was up-regulated. To define the link between mitochondrial dysfunction and disuse muscle atrophy we unloaded mice overexpressing PGC-1α. Transgenic PGC-1α animals did not show metabolic alteration during unloading, preserving muscle size through the reduction of autophagy and proteasome degradation. Our results indicate that mitochondrial dysfunction plays a major role in disuse atrophy and that compounds inducing PGC-1α expression could be useful to treat/prevent muscle atrophy.

Can gas replace protein function? CO abrogates the oxidative toxicity of myoglobin.

Outside their cellular environments, hemoglobin (Hb) and myoglobin (Mb) are known to wreak oxidative damage. Using haptoglobin (Hp) and hemopexin (Hx) the body defends itself against cell-free Hb, yet mechanisms of protection against oxidative harm from Mb are unclear. Mb may be implicated in oxidative damage both within the myocyte and in circulation following rhabdomyolysis. Data from the literature correlate rhabdomyolysis with the induction of Heme Oxygenase-1 (HO-1), suggesting that either the enzyme or its reaction products are involved in oxidative protection. We hypothesized that carbon monoxide (CO), a product, might attenuate Mb damage, especially since CO is a specific ligand for heme iron. Low density lipoprotein (LDL) was chosen as a substrate in circulation and myosin (My) as a myocyte component. Using oxidation targets, LDL and My, the study compared the antioxidant potential of CO in Mb-mediated oxidation with the antioxidant potential of Hp in Hb-mediated oxidation. The main cause of LDL oxidation by Hb was found to be hemin which readily transfers from Hb to LDL. Hp prevented heme transfer by sequestering hemin within the Hp-Hb complex. Hemin barely transferred from Mb to LDL, and oxidation appeared to stem from heme iron redox in the intact Mb. My underwent oxidative crosslinking by Mb both in air and under N2. These reactions were fully arrested by CO. The data are interpreted to suit several circumstances, some physiological, such as high muscle activity, and some pathological, such as rhabdomyolysis, ischemia/reperfusion and skeletal muscle disuse atrophy. It appear that CO from HO-1 attenuates damage by temporarily binding to deoxy-Mb, until free oxygen exchanges with CO to restore the equilibrium.

Skeletal Muscle Disuse Atrophy Is Not Attenuated by Dietary Protein Supplementation in Healthy Older Men

Short successive periods of muscle disuse, due to injury or illness, can contribute significantly to the loss of muscle mass with aging (sarcopenia). It has been suggested that increasing the protein content of the diet may be an effective dietary strategy to attenuate muscle disuse atrophy. We hypothesized that protein supplementation twice daily would preserve muscle mass during a short period of limb immobilization. Twenty-three healthy older (69 ± 1 y) men were subjected to 5 d of one-legged knee immobilization by means of a full-leg cast with (PRO group; n = 11) or without (CON group; n = 12) administration of a dietary protein supplement (20.7 g of protein, 9.3 g of carbohydrate, and 3.0 g of fat) twice daily. Two d prior to and immediately after the immobilization period, single-slice computed tomography scans of the quadriceps and single-leg 1 repetition maximum strength tests were performed to assess muscle cross-sectional area (CSA) and leg muscle strength, respectively. Additionally, muscle biopsies were collected to assess muscle fiber characteristics as well as mRNA and protein expression of selected genes. Immobilization decreased quadriceps' CSAs by 1.5 ± 0.7% (P < 0.05) and 2.0 ± 0.6% (P < 0.05), and muscle strength by 8.3 ± 3.3% (P < 0.05) and 9.3 ± 1.6% (P < 0.05) in the CON and PRO groups, respectively, without differences between groups. Skeletal muscle myostatin, myogenin, and muscle RING-finger protein-1 (MuRF1) mRNA expression increased following immobilization in both groups (P < 0.05), whereas muscle atrophy F-box/atrogen-1 (MAFBx) mRNA expression increased in the PRO group only (P < 0.05). In conclusion, dietary protein supplementation (∼20 g twice daily) does not attenuate muscle loss during short-term muscle disuse in healthy older men. This trial was registered at clinicaltrials.gov as NCT01588808.

Aging Affects the Transcriptional Regulation of Human Skeletal Muscle Disuse Atrophy

Important insights concerning the molecular basis of skeletal muscle disuse-atrophy and aging related muscle loss have been obtained in cell culture and animal models, but these regulatory signaling pathways have not previously been studied in aging human muscle. In the present study, muscle atrophy was induced by immobilization in healthy old and young individuals to study the time-course and transcriptional factors underlying human skeletal muscle atrophy. The results reveal that irrespectively of age, mRNA expression levels of MuRF-1 and Atrogin-1 increased in the very initial phase (2–4 days) of human disuse-muscle atrophy along with a marked reduction in PGC-1α and PGC-1β (1–4 days) and a ∼10% decrease in myofiber size (4 days). Further, an age-specific decrease in Akt and S6 phosphorylation was observed in young muscle within the first days (1–4 days) of immobilization. In contrast, Akt phosphorylation was unchanged in old muscle after 2 days and increased after 4 days of immobilization. Further, an age-specific down-regulation of MuRF-1 and Atrogin-1 expression levels was observed following 2 weeks of immobilization, along with a slowing atrophy response in aged skeletal muscle. Neither the immediate loss of muscle mass, nor the subsequent age-differentiated signaling responses could be explained by changes in inflammatory mediators, apoptosis markers or autophagy indicators. Collectively, these findings indicate that the time-course and regulation of human skeletal muscle atrophy is age dependent, leading to an attenuated loss in aging skeletal muscle when exposed to longer periods of immobility-induced disuse.