Lack of muscle stem cell proliferation and myocellular hypertrophy in sIBM patients following blood-flow restricted resistance training

Sporadic inclusion body myositis (sIBM) is characterised by skeletal muscle inflammation, progressive muscle loss and weakness, which is largely refractory to immunosuppressive treatment. Low-load blood-flow restricted (BFR) training has been shown to evoke gains in myofibre cross sectional area (mCSA) in healthy adults. This could partially be due to the activation and integration of muscle satellite cells (SC) resulting in myonuclei addition. Consequently, this study investigated the effect of 12-weeks lower limb low-load BFR resistance training in sIBM patients on SC and myonuclei content, myofibre size and capillarization. Muscle biopsies from sIBM patients randomised to 12-weeks of low-load BFR resistance training (n = 11) or non-exercising controls (CON) (n = 9) were analysed for SC and myonuclei content, myofibre size and capillarization using three-colour immunofluorescence microscopy and computerised quantification procedures. No between-group differences (time-by-group interactions) or within-groups changes were observed for resident SCs (Pax7⁺/Six1⁺), proliferating SCs (Pax7⁺/ Ki67⁺), myonuclei (Six1⁺), type 1 mCSA or capillary number (CD31⁺). However, a time-by-group interaction for type 2 mCSA was observed (p = 0.04). Satellite cell content, myonuclei number, mCSA and capillary density remained unaffected following 12-weeks low-load BFR resistance training, indicating limited myogenic capacity and satellite cell plasticity in long-term sIBM patients.     

Fatigue and recovery from dynamic contractions in men and women differ for arm and leg muscles

Introduction: Whether there is a gender difference in fatigue and recovery from maximal velocity fatiguing contractions and across muscles is not understood. Methods: Sixteen men and 19 women performed 90 isotonic contractions at maximal voluntary shortening velocity (maximal velocity concentric contractions, MVCC) with the elbow flexor and knee extensor muscles (separate days) at a load equivalent to 20% maximal voluntary isometric contraction (MVIC). Results: Power (from MVCCs) decreased similarly for men and women for both muscles (P > 0.05). Men and women had similar declines in MVIC of elbow flexors, but men had greater reductions in knee extensor MVIC force and MVIC electromyogram activity than women (P < 0.05). The decline in MVIC and power was greater, and force recovery was slower for the elbow flexors compared with knee extensors. Conclusions: The gender difference in muscle fatigue often observed during isometric tasks was diminished during fast dynamic contractions for upper and lower limb muscles. Muscle Nerve 48 : 436–439, 2013     

Mechanosensitivity may be enhanced in skeletal muscles of spinal cord–injured versus able‐bodied men

We investigated the effects of an acute bout of neuromuscular electrical stimulation–induced resistance exercise (NMES‐RE) on intracellular signaling pathways involved in translation initiation and mechanical loading–induced muscle hypertrophy in spinal cord–injured (SCI) versus able‐bodied (AB) individuals. AB and SCI individuals completed 90 isometric knee extension contractions at 30% of maximum voluntary or evoked contraction, respectively. Muscle biopsies were collected before, and 10 and 60 min after NMES‐RE. Protein levels of α7‐ and β1‐integrin, phosphorylated and total GSK‐3α/β, S6K1, RPS6, 4EBP1, and FAK were assessed by immunoblotting. SCI muscle appears to be highly sensitive to muscle contraction even several years after the injury, and in fact it may be more sensitive to mechanical stress than AB muscle. Heightened signaling associated with muscle mechanosensitivity and translation initiation in SCI muscle may be an attempted compensatory response to offset elevated protein degradation in atrophied SCI muscle. Muscle Nerve 50: 599–601, 2014     

Knee extensor strength, activation, and size in very elderly people following strength training.

Muscle strength, activation, and size were studied in 11 very elderly subjects (8 women and 3 men; age range, 85-97 years) who completed 12 weeks of strength training of the knee extensor muscles. Training increased the maximum amount of weight that could be lifted once (134%; P < 0.05) and maximum voluntary isometric strength, measured as both force recorded at the ankle with the knee flexed 90 degrees (17%, ns) and as torque with the knee flexed 60 degrees (37%; P < 0.05). Anatomical lean quadriceps cross-sectional area (LCSA) measured at midthigh using magnetic resonance imaging increased from 27.5 +/- 9.6 cm2 to 30.2 +/- 10.0 cm2 (9.8%; P < 0. 05) after training. Both before and after training, isometric strength was closely related to LCSA, but training resulted in no significant change in muscle force per unit area of quadriceps muscle. Using the twitch interpolation technique, muscle activation during a maximal voluntary isometric contraction was shown to be incomplete in all subjects before training (ranging from 69% to 93%) and was not significantly increased after training. An increase in skeletal muscle mass may have important functional and metabolic benefits for very elderly people.     

Stretch‐activated signaling is modulated by stretch magnitude and contraction

Introduction: Stretch therapy is commonly utilized to prevent shortening maladaptation of skeletal muscle. Stretch in combination with isometric contraction prevents shortening, but the signaling mechanisms are not understood. Methods: Using a soleus tenotomy + stretch rat model, the phosphorylation–activation of mechanosensitive kinases (Akt, p70^S6K, p38 MAPK, and ERK1/2) were measured for various stretch magnitudes, set relative to optimal soleus length (Lo). Results: The kinases were not activated by passive stretch until it exceeded the normal physiological range. Stretch + isometric contraction resulted in relatively strong phosphorylation, even at short lengths. Conclusions: Whereas passive stretch results in kinase phosphorylation only during extreme lengthening, isometric contraction generated pronounced phosphorylation of kinases at Lo and Lo + 25%, indicating stimulation of pathways that lead to the preservation or increase of muscle length. Understanding the effects of passive and active stretch with respect to Lo and contraction is essential for predicting therapeutic outcomes and influencing optimal muscle length. Muscle Nerve 49 : 98–107, 2014     

Electrical stimulation influences satellite cell differentiation after sciatic nerve crush injury in rats

Introduction: Electrical stimulation is often used to prevent muscle atrophy and preserve contractile function, but its effects on the satellite cell population after nerve injury are not well understood. In this study we aimed to determine whether satellite cell differentiation is affected by electrical stimulation after nerve crush. Methods: The sciatic nerves of Sprague‐Dawley (SD) rats were crushed. Half of the injured rats received daily electrical stimulation of the gastrocnemius muscle, and the others did not. Tests for detecting paired box protein 7 (Pax7), myogenic differentiation antigen (MyoD), embryonic myosin heavy chain (eMyHC), and force production were performed 2, 4, and 6 weeks after injury. Results: More Pax7⁺/MyoD⁺ nuclei in stimulated muscles were observed than in non‐stimulated muscles. eMyHC expression was elevated in stimulated muscles and correlated positively with enhanced force production. Conclusions: Increased satellite cell differentiation is correlated with preserved muscle function in response to electrical stimulation after nerve injury. Muscle Nerve 51: 400–411, 2015     

Invasive fibroblasts: Fundamental difference between sporadic inclusion body myositis and polymyositis

Introduction: Sporadic inclusion body myositis (sIBM) is a progressive disease that leads to extensive muscle weakness. The aim of this study was to determine whether the number and distribution of fibroblasts differ in sIBM when compared with polymyositis. Methods: Immunofluorescence double labeling was performed on 35 biopsies with antibodies directed against perlecan and CD90, procollagen I, CD34, and CD105. In addition, nonserial ultrathin sections were studied from 3 biopsies of each condition. Results: Fibroblasts expressing CD90 and CD34 accumulated in the endomysial compartment in polymyositis and sIBM. In addition, cells expressing CD90 were found beneath the basal lamina in both conditions. At the ultrastructural level in polymyositis, fibroblasts invaded the myofiber, with focal destruction of the basement membrane. In sIBM, by contrast, invasive fibroblasts were ensheathed by the intact myofiber basement membrane. Conclusions: The impact of intruding fibroblasts on satellite cells remains to be established. Muscle Nerve 49 : 175–180, 2014     

Behavior of fascicles and the myotendinous junction of human medial gastrocnemius following eccentric strength training

This study is the first in which measurements of thickness, fascicle angle and length, and tendon elongation were combined to examine the impact of eccentric strength training on both muscle architecture and tendinous structures. Eighteen healthy male subjects were divided into an eccentric strength training group (n = 10) and a control group (n = 8). The training program consisted of 18 sessions of eccentric exercises over a 7‐week period. All subjects were tested at baseline and after the last training session. Using ultrasound imaging, the fascicle angle and length and thickness of the medial gastrocnemius (MG) were analyzed at rest (i.e., θ_p, Fl_p, and t_p, respectively), at 50% of maximal voluntary contraction (MVC) (i.e., θ₅₀, Fl₅₀, and t₅₀, respectively), and during MVC (i.e., θ₁₀₀, Fl₁₀₀, and t₁₀₀, respectively). Tendon elongation (TE) was measured by tracking the proximal displacement of the myotendinous junction of the MG during ramp isometric contraction. During ramp isometric contraction, the slope of the load–deformation relationship of the gastrocnemius tendon above 50% MVC was defined as an index of stiffness. After training, muscle thickness and fascicle angle increased significantly at rest and during contraction, whereas fascicle length increased at rest and did not change during contraction. Furthermore, the stiffness of the gastrocnemius tendon increased significantly. The results suggest that the behavior of muscle architecture and tendon mechanical properties are affected differently by strength training. Muscle Nerve, 2009     

Axonal excitability during ischemia in MELAS

Introduction: We investigated muscle activation strategy and performance of knee extensor and flexor muscles in children and adults with generalized joint hypermobility (GJH) and compared them with controls. Methods: Muscle activation, torque steadiness, electromechanical delay, and muscle strength were evaluated in 39 children and 36 adults during isometric knee extension and flexion. Subjects performed isometric maximum contractions, submaximal contractions at 25% maximum voluntary contraction (MVC), and explosive contractions. Results: Agonist activation was reduced, and coactivation ratio was greater in GJH during knee flexion compared with controls. Torque steadiness was impaired in adults with GJH during knee flexion. No effect of GJH was found on muscle strength or electromechanical delay. Correlation analysis revealed an association between GJH severity and function in adults. Conclusions: The results indicate that muscle activation strategy and quality of force control were significantly affected in adults with GJH during knee flexion, whereas only muscle activation strategy was affected in children with GJH. Muscle Nerve 48:762–769, 2013     

Myonuclear domains in muscle adaptation and disease.

Adult skeletal muscle fibers are among the few cell types that are truly multinucleated. Recently, evidence has accumulated supporting a role for the modulation of myonuclear number during muscle remodeling in response to injury, adaptation, and disease. These studies have demonstrated that muscle hypertrophy is associated with, and is dependent on, the addition of newly formed myonuclei via the fusion of myogenic cells to the adult myofiber, whereas muscle atrophy and disease appear to be associated with the loss of myonuclei, possibly through apoptotic-like mechanisms. Moreover, these studies also have demonstrated that myonuclear domain size, i. e., the amount of cytoplasm per myonucleus, is unchanged following the acute phase of hypertrophy but is reduced following atrophy. Together these data demonstrate that modulation of myonuclear number or myonuclear domain size (or both) is a mechanism contributing to the remodeling of adult skeletal muscle in response to alterations in the level of normal neuromuscular activity.     

Satellite cell pool expansion is affected by skeletal muscle characteristics

Introduction: We investigated changes in satellite cell (SC) pool size after an acute bout of strenuous exercise and evaluated the influence of baseline SC count and fiber type. Methods: Participants completed a downhill running (DHR) intervention (5 × 8 min, 2‐min rest; 80% VO _2max; ?10% gradient). Muscle biopsies were taken 7 days before VO _2max and 7–9 days after the DHR intervention. Delayed‐onset muscle soreness (DOMS) and creatine kinase activity (CK) were measured on days 1, 2, 7, and 9 post‐DHR. SCs were identified by Pax7 and laminin staining. Relative distribution of MHC isoforms was determined by electrophoresis. Results: DOMS and CK peaked on day 1 post‐DHR (P < 0.01). The SC pool increased (26%) after DHR (P = 0.005). SCs/total myonuclei after recovery correlated with baseline SCs (r = 0.979, P = 0.003) and VO _2max (r = 0.956, P = 0.011), whereas change in SC pool (Pax7⁺ cells/total myonuclei: recovery minus baseline) tended to correlate with percent MHC II (r = 0.848; P = 0.06). Conclusion: Interindividual physiological characteristics affect SC pool expansion after a single bout of DHR and are influenced by VO _2max. Muscle Nerve, 2013     

Inhibition of microRNA-92a increases blood vessels and satellite cells in skeletal muscle but does not improve duchenne muscular dystrophy–related phenotype in mdx mice

Introduction: The vasculature and blood flow in muscle are perturbed in Duchenne muscular dystrophy (DMD) and its mdx mouse model. MicroRNA-92a (miR-92a) is enriched in endothelial cells, especially during ischemic injury. Methods: Because antagonizing miR-92a was shown to result in increased proliferation and migration of endothelial cells and recovery from ischemia, we assessed the effects of Antagomir-92a in vitro in muscle stem cell culture and in vivo in mdx mice. Results: miR-92a was found to be highly expressed in muscle endothelial cells and satellite cells. Treatment with Antagomir-92a increased capillary density and tissue perfusion, which was accompanied by an increase in satellite cells. However, Antagomir-92a–treated mdx mice showed no histological improvement and had worse muscle function. Antagomir-92a suppressed myogenic differentiation in satellite cell culture. Discussion: AntagomiR-92a improves the vasculature but not the muscle in mdx mice, possibly due to its side effects on satellite cell differentiation. Muscle Nerve 59:594–594, 2019     

Regenerative capacity and the number of satellite cells in soleus muscles of normal and mdx mice

Satellite cells are potential myogenic cells that participate in repair and growth of muscle fibres. In this investigation, the change in the number of satellite cells following severe muscle damage was monitored in soleus muscle of age-matched mdx and C57Bl/10 mice. Satellite cells were identified immunohistochemically in the light microscope by their association with a recently described marker protein, M-cadherin, and their location between the muscle fibre's sarcolemma and the surrounding basal lamina. In cross-sections of untreated soleus muscle of C57Bl/10 mice at 11-14. 5 months of age, nuclei of M-cadherin positive satellite cells on average amounted to 3.4% of the total number of myonuclei. Surprisingly, significantly higher numbers of satellite cell nuclei, both in absolute numbers (mean 24+/-11 versus 40+/-11 satellite cells per section) and relative to the total number of myonuclei (5. 3%), were found in similarly aged animals in which severe muscle damage had been inflicted 3-6 months before. Cross-sectional area, muscle tissue area and myonuclei counts had recovered to control values. In untreated muscles of age-matched mdx mice satellite cell counts were not different (2.7% of myonuclei) from C57Bl/10 mice. However, regeneration showed marked deficits, as there was a loss of about 36% total cross-sectional area, about 48% total muscle fibre area and about 43% myonuclei per section compared to the untreated mdx muscles. Furthermore, the absolute number of satellite cells decreased from 20+/-11 to 12+/-8 per section. The relative number of satellite cell nuclei remained comparable to, but did not exceed, the undamaged muscles. The poor recovery of muscle and the missing post-regeneration rise in satellite cell numbers may indicate the reproductive limits of the satellite pool.     

Impaired regeneration in LGMD2A supported by increased PAX7‐positive satellite cell content and muscle‐specific microrna dysregulation

Introduction: Recent in vitro studies suggest that CAPN3 deficiency leads initially to accelerated myofiber formation followed by depletion of satellite cells (SC). In normal muscle, up‐regulation of miR‐1 and miR‐206 facilitates transition from proliferating SCs to differentiating myogenic progenitors. Methods: We examined the histopathological stages, Pax7 SC content, and muscle‐specific microRNA expression in biopsy specimens from well‐characterized LGMD 2A patients to gain insight into disease pathogenesis. Results: Three distinct stages of pathological changes were identified that represented the continuum of the dystrophic process from prominent inflammation with necrosis and regeneration to prominent fibrosis, which correlated with age and disease duration. Pax7‐positive SCs were highest in the fibrotic group and correlated with down‐regulation of miR‐1, miR‐133a, and miR‐206. Conclusions: These observations, and other published reports, are consistent with microRNA dysregulation leading to inability of Pax7‐positive SCs to transit from proliferation to differentiation. This results in impaired regeneration and fibrosis. Muscle Nerve 47: 731–739, 2013     

Power spectral changes of the superimposed M wave during isometric voluntary contractions of increasing strength

Introduction: We examined the power spectral changes of the compound muscle action potential (M wave) evoked during isometric contractions of increasing strength. Methods: Surface electromyography (sEMG) of the vastus lateralis and medialis was recorded from 20 volunteers who performed 4‐s step‐wise isometric contractions of different intensities. A maximal M wave was elicited by a single stimulus to the femoral nerve and superimposed on the voluntary contractions. The spectral characteristics (Fmean and Fmedian) of sEMG and M‐wave signals were calculated. Results: M‐wave spectral indicators increased systematically with contraction intensity up to 60% MVC and then leveled off at higher forces. Over the 10–60% MVC range, the increase in spectral indicators was 3 times higher for M waves (36%) than for sEMG (12%). Conclusions: The consistent increase in M‐wave spectral characteristics with force is due to the fact that the number of motor units recruited by the superimposed supramaximal stimulus is approximately stable. Muscle Nerve 51: 580–591, 2015     

Motor unit control strategies of endurance‐ versus resistance‐trained individuals

Introduction: We examined motor unit (MU) control properties of resistance‐trained (RT) and endurance‐trained (ET) individuals. Methods: Five RT (age 25 ± 4 years) and 5 ET (age 19 ± 1 years) subjects participated. Surface electromyography (EMG) data were recorded from the vastus lateralis during isometric trapezoid muscle actions at 40% and 70% of maximal voluntary contraction. Decomposition and wavelet transform techniques were applied to the EMG signals to extract the firing events of single MUs and EMG intensity across the frequency spectrum. Results: There were significant differences between RT and ET for the mean MU firing rate and derecruitment threshold versus recruitment threshold relationships and EMG intensity at various wavelet bands during the linearly increasing, steady force, and linearly decreasing segments of the trapezoid contraction. Conclusions: MU behavior is altered as a function of training status and is likely the result of differences in the physical properties of the MU. Muscle Nerve 52 : 832–843, 2015     

Immunohistochemical analysis of the distribution of MyoD1 in muscle biopsies of primary myopathies and neurogenic atrophy

The expression of the myogenic determination gene MyoD1 plays a primary role in the commitment of primitive mesenchymal cells to a striated muscle lineage and is down-regulated during later stages of differentiation. To determine the potential role of this gene in myopathic conditions, we examined its expression by means of immunohistochemical analysis, using a series of muscle biopsies from 14 patients with a variety of primary myopathies and neurogenic disorders. Utilizing the avidin-biotin-complex technique, cryostat sections were stained with monoclonal antibody 5.8 A, which we have previously described as having a high level of specificity for tumors with rhabdomyoblastic differentiation. Of special interest was the observation in 4 of 8 cases of neurogenic atrophy of varying levels of cytoplasmic positivity of muscle fibers, appearing to correlate with their degree of atrophy, in addition to weak nuclear staining. Muscle biopsies from 2 patients with Duchenne's muscular dystrophy and 2 patients with autoimmune inflammatory myopathies demonstrated various levels of nuclear positivity in scattered foci that appeared to correlate with areas of regeneration. A biopsy from a single case of neurogenic atrophy secondary to infantile spinal muscular atrophy (Werdnig-Hoffmann's disease) demonstrated diffuse but relatively weak staining of myofiber nuclei, in contrast to sections of normal striated muscle and muscle biopsies from patients with unexplained myoglobinuria, which exhibited only minimal amounts of staining. These data are compatible with observations that MyoD1 expression is related to electrical activity and muscle regeneration.Supported by Cancer Center Support (CORE) grant CA21765 and grant CA23099 from the National Cancer Institute, and by the American Lebanese-Syrian Associated Charities (ALSAC) Presented at the Annual Meeting of the United States and Canadian Academy of Pathology, New Orleans, LA 1993 (Mod Pathol 6: 120A, 1993)     

Age‐associated changes in muscle activity during isometric contraction

Introduction: We investigated the effect of age on the complexity of muscle activity and the variance in the force of isometric contraction. Methods: Surface electromyography (sEMG) from biceps brachii muscle and force of contraction were recorded from 96 subjects (20–70 years of age) during isometric contractions. Results: There was a reduction in the complexity of sEMG associated with aging. The relationship of age and complexity was approximated using a bilinear fit, with the average knee point at 45 years. There was an age‐associated increase in the coefficient of variation (CoV) of the force of muscle contraction, and this increase was correlated with the decrease in complexity of sEMG (r² = 0.76). Conclusions: There was an age‐associated increase in CoV and also a reduction in the complexity of sEMG. The correlation between these 2 factors can be explained based on the age‐associated increase in motor unit density. Muscle Nerve 47: 545–549, 2013     

Intersession reliability of the interpolated twitch technique applied during isometric,concentric, and eccentric actions of the human knee extensor muscles

Introduction: Although it has been shown that voluntary activation (%VA) of the knee extensors during isometric contractions can be reliably assessed with the interpolated twitch technique, little is known about the reliability of %VA during concentric and eccentric muscle actions. Therefore, relative and absolute intersession reliability of quadriceps muscle's %VA during different contraction modes was determined. Methods: After a familiarization session, 21 participants (17 males, 25 ± 2 yrs) completed two testing sessions. Paired supramaximal electrical stimuli were administered to the femoral nerve during isometric, concentric, eccentric MVCs, and at rest to assess %VA (stimuli were applied at 70° knee flexion). Results and discussion: Data indicate that %VA of the knee extensors can be reliably measured during isometric [intraclass correlation coefficient (ICC) = 0.89, coefficient of variation (CV) = 4.1%], concentric (ICC = 0.87, CV = 6.6%), and eccentric muscle actions (ICC = 0.86, CV = 7.0%). Muscle Nerve 56 : 324–327, 2017     

Knee extensor fatigue resistance of young and older men and women performing sustained and brief intermittent isometric contractions

Introduction: Susceptibility to muscle fatigue during aging could depend on muscle activation patterns. Methods: Young (mean age, 22 years) and older (mean age 70 years) men and women completed two fatigue tests of knee extensor muscles using voluntary and electrically stimulated contractions. Results: Older subjects displayed a shift to the left of the torque‐frequency relationship and held a sustained voluntary isometric contraction at 50% maximal strength for significantly longer than young (P < 0.001). Young and old showed similar fatigue during electrically induced, intermittent isometric contractions (1‐s on, 1‐s off for 2 min), but women fatigued less than men (P = 0.001). Stronger muscles fatigued more quickly, and slower contractile properties were associated with longer sustained contractions. Conclusions: The slowing and weakness of older muscle was associated with superior fatigue resistance during sustained isometric contractions. Young and old showed similar fatigue following a series of brief, intermittent contractions, but women fatigued less than men. Muscle Nerve 50 : 393–400, 2014