Skeletal Muscle Force Research Articles

JAK inhibition with tofacitinib rapidly increases contractile force in human skeletal muscle.

Reduction in muscle contractile force associated with many clinical conditions incurs serious morbidity and increased mortality. Here, we report the first evidence that JAK inhibition impacts contractile force in normal human muscle. Muscle biopsies were taken from patients who were randomized to receive tofacitinib (n = 16) or placebo (n = 17) for 48 h. Single-fiber contractile force and molecular studies were carried out. The contractile force of individual diaphragm myofibers pooled from the tofacitinib group (n = 248 fibers) was significantly higher than those from the placebo group (n = 238 fibers), with a 15.7% greater mean maximum specific force (P = 0.0016). Tofacitinib treatment similarly increased fiber force in the serratus anterior muscle. The increased force was associated with reduced muscle protein oxidation and FoxO-ubiquitination-proteasome signaling, and increased levels of smooth muscle MYLK. Inhibition of MYLK attenuated the tofacitinib-dependent increase in fiber force. These data demonstrate that tofacitinib increases the contractile force of skeletal muscle and offers several underlying mechanisms. Inhibition of the JAK-STAT pathway is thus a potential new therapy for the muscle dysfunction that occurs in many clinical conditions.

The mitochondrial-targeted peptide therapeutic elamipretide improves cardiac and skeletal muscle function during aging without detectable changes in tissue epigenetic or transcriptomic age.

Aging-related decreases in cardiac and skeletal muscle function are strongly associated with various comorbidities. Elamipretide (ELAM), a novel mitochondrial-targeted peptide, has demonstrated broad therapeutic efficacy in ameliorating disease conditions associated with mitochondrial dysfunction across both clinical and pre-clinical models. ELAM is proposed to restore mitochondrial bioenergetic function by stabilizing inner membrane structure and increasing oxidative phosphorylation coupling and efficiency. Although ELAM treatment effectively attenuates physiological declines in multiple tissues in rodent aging models, it remains unclear whether these functional improvements correlate with favorable changes in molecular biomarkers of aging. Herein, we investigated the impact of 8-week ELAM treatment on pre- and post-measures of C57BL/6J mice frailty, skeletal muscle, and cardiac muscle function, coupled with post-treatment assessments of biological age and affected molecular pathways. We found that health status, as measured by frailty index, cardiac strain, diastolic function, and skeletal muscle force are significantly diminished with age, with skeletal muscle force changing in a sex-dependent manner. Conversely, ELAM mitigated frailty accumulation and was able to partially reverse these declines, as evidenced by treatment-induced increases in cardiac strain and muscle fatigue resistance. Despite these improvements, we did not detect statistically significant changes in gene expression or DNA methylation profiles indicative of molecular reorganization or reduced biological age in most ELAM-treated groups. However, pathway analyses revealed that ELAM treatment showed pro-longevity shifts in gene expression such as upregulation of genes involved in fatty acid metabolism, mitochondrial translation and oxidative phosphorylation, and downregulation of inflammation. Together, these results indicate that ELAM treatment is effective at mitigating signs of sarcopenia and heart failure in an aging mouse model, but that these functional improvements occur independently of detectable changes in epigenetic and transcriptomic age. Thus, some age-related changes in function may be uncoupled from changes in molecular biological age.

Molecular determinants of skeletal muscle force loss in response to 5days of dry immersion in human.

Astronauts in Earth's orbit experience microgravity, resulting in a decline of skeletal muscle mass and function. On Earth, models simulating microgravity have shown that the extent of the loss in muscle force is greater than the loss in muscle mass. The reasons behind this disproportionate loss of muscle force are still poorly understood. In the present study, we hypothesize that alongside the loss in skeletal muscle mass, modifications in the expression profile of genes encoding critical determinants of resting membrane potential, excitation-contraction coupling and Ca2+ handling contribute to the decline in skeletal muscle force. Healthy male volunteers (n=18) participated in a 5-day dry immersion (DI) study, an Earth-based model of simulated microgravity. Muscle force measurement and MRI analysis of the cross-sectional area of thigh muscles were performed before and after DI. Biopsies of the vastus lateralis skeletal muscle performed before and after DI were used for the determination Ca2+ properties of isolated muscle fibres, molecular and biochemical analyses. The extent of the decline in force, measured as maximal voluntary contraction of knee extensors (-11.1%, P<0.01) was higher than the decline in muscle mass (-2.5%, P<0.01). The decline in muscle mass was molecularly supported by a significant repression of the anabolic IGF-1/Akt/mTOR pathway (-19.9% and -40.9% in 4E-BP1 and RPS6 phosphorylation, respectively), a transcriptional downregulation of the autophagy-lysosome pathway and a downregulation in the mRNA levels of myofibrillar protein slow isoforms. At the single fibre level, biochemical and tension-pCa curve analyses showed that the loss in force was independent of fibre type (-11% and -12.3% in slow and fast fibres, respectively) and Ca2+ activation properties. Finally, we showed a significant remodelling in the expression of critical players of resting membrane potential (aquaporin 4: -24.9%, ATP1A2: +50.4%), excitation-contraction coupling (CHRNA1: +75.1%, CACNA2D1: -23.5%, JPH2: -24.2%, TRDN: -15.6%, S100A1: +27.2%), and Ca2+ handling (ATP2A2: -32.5%, CASQ1: -15%, ORAI1: -36.2%, ATP2B1: -19.1%). These findings provide evidence that a deregulation in the expression profile of critical molecular determinants of resting membrane potential, excitation-contraction coupling, and Ca2+ handling could be involved in the loss of muscle force induced by DI. They also provide the paradigm for the understanding of muscle force loss during prolonged bed rest periods as those encountered in intensive care unit.

Impact of Eccentric Exercise Interventions with Small and Large Ranges of Motion on Rat Skeletal Muscle Tissue and Muscle Force Production.

Eccentric training induces greater hypertrophy while causing more muscle damage than concentric training. This study examined the effects of small-range eccentric contractions (SR-ECCs) and large-range eccentric contractions (LR-ECCs) on muscle morphology, contractility, and damage in rats. Thirty male Fischer 344 rats were divided into five groups: small-range ECC single-bout (SR-ECCSB, n = 4), large-range ECC single-bout (LR-ECCSB, n = 4), SR-ECC intervention (SR-ECCIntv, n = 7), LR-ECC intervention (LR-ECCIntv, n = 8), and control (Cont, n = 7). These groups underwent transcutaneous electrical stimulation involving 80 ECCs twice a week for four weeks. The results indicated that the LR-ECCSB group had more Evans blue dye-positive fibers than other groups. The SR-ECCIntv group showed no increase in the mean myofiber cross-sectional area. However, Pax7+ and Ki67+ cells significantly increased in both ECCIntv groups compared to the Cont group, and the connective tissue area was significantly greater in the LR-ECCIntv than in others. Muscle force was lower in both ECCIntv groups compared to the Cont group. These findings suggest that SR-ECC intervention may induce a smaller increase in the number of fibers with a large myofiber cross-sectional area and satellite cell proliferation with less muscle damage and myofibrosis compared to LR-ECCs.

Integrated Wearable System for Monitoring Skeletal Muscle Force of Lower Extremities.

Continuous monitoring of lower extremity muscles is necessary, as the muscles support many human daily activities, such as maintaining balance, standing, walking, running, and jumping. However, conventional electromyography and physiological cross-sectional area methods inherently encounter obstacles when acquiring precise and real-time data pertaining to human bodies, with a notable lack of consideration for user comfort. Benefitting from the fast development of various fabric-based sensors, this paper addresses these current issues by designing an integrated smart compression stocking system, which includes compression garments, fabric-embedded capacitive pressure sensors, an edge control unit, a user mobile application, and cloud backend. The pipeline architecture design and component selection are discussed in detail to illustrate a comprehensive user-centered STIMES design. Twelve healthy young individuals were recruited for clinical experiments to perform maximum voluntary isometric ankle plantarflexion contractions. All data were simultaneously collected through the integrated smart compression stocking system and a muscle force measurement system (Humac NORM, software version HUMAC2015). The obtained correlation coefficients above 0.92 indicated high linear relationships between the muscle torque and the proposed system readout. Two-way ANOVA analysis further stressed that different ankle angles (p = 0.055) had more important effects on the results than different subjects (p = 0.290). Hence, the integrated smart compression stocking system can be used to monitor the muscle force of the lower extremities in isometric mode.

The effect of muscle ultrastructure on the force, displacement and work capacity of skeletal muscle.

Skeletal muscle powers animal movement through interactions between the contractile proteins, actin and myosin. Structural variation contributes greatly to the variation in mechanical performance observed across muscles. In vertebrates, gross structural variation occurs in the form of changes in the muscle cross-sectional area : fibre length ratio. This results in a trade-off between force and displacement capacity, leaving work capacity unaltered. Consequently, the maximum work per unit volume-the work density-is considered constant. Invertebrate muscle also varies in muscle ultrastructure, i.e. actin and myosin filament lengths. Increasing actin and myosin filament lengths increases force capacity, but the effect on muscle fibre displacement, and thus work, capacity is unclear. We use a sliding-filament muscle model to predict the effect of actin and myosin filament lengths on these mechanical parameters for both idealized sarcomeres with fixed actin : myosin length ratios, and for real sarcomeres with known filament lengths. Increasing actin and myosin filament lengths increases stress without reducing strain capacity. A muscle with longer actin and myosin filaments can generate larger force over the same displacement and has a higher work density, so seemingly bypassing an established trade-off. However, real sarcomeres deviate from the idealized length ratio suggesting unidentified constraints or selective pressures.

GDF5 as a rejuvenating treatment for age-related neuromuscular failure.

Sarcopenia involves a progressive loss of skeletal muscle force, quality and mass during ageing, which results in increased inability and death; however, no cure has been established thus far. Growth differentiation factor 5 (GDF5) has been described to modulate muscle mass maintenance in various contexts. For our proof of concept, we overexpressed GDF5 by AAV vector injection in tibialis anterior muscle of adult aged (20 months) mice and performed molecular and functional analysis of skeletal muscle. We analysed human vastus lateralis muscle biopsies from adult young (21-42 years) and aged (77-80 years) donors, quantifying the molecular markers modified by GDF5 overexpression in mouse muscle. We validated the major effects of GDF5 overexpression using human immortalized myotubes and Schwann cells. We established a preclinical study by treating chronically (for 4 months) aged mice using recombinant GDF5 protein (rGDF5) in systemic administration and evaluated the long-term effect of this treatment on muscle mass and function. Here, we demonstrated that GDF5 overexpression in the old tibialis anterior muscle promoted an increase of 16.5% of muscle weight (P = 0.0471) associated with a higher percentage of 5000-6000 µm2 large fibres (P = 0.0211), without the induction of muscle regeneration. Muscle mass gain was associated with an amelioration of 26.8% of rate of force generation (P = 0.0330) and better neuromuscular connectivity (P = 0.0098). Moreover, GDF5 overexpression preserved neuromuscular junction morphology (38.5% of nerve terminal area increase, P < 0.0001) and stimulated the expression of reinnervation-related genes, in particular markers of Schwann cells (fold-change 3.19 for S100b gene expression, P = 0.0101). To characterize the molecular events induced by GDF5 overexpression during ageing, we performed a genome-wide transcriptomic analysis of treated muscles and showed that this factor leads to a 'rejuvenating' transcriptomic signature in aged mice, as 42% of the transcripts dysregulated by ageing reverted to youthful expression levels upon GDF5 overexpression (P < 0.05). Towards a preclinical approach, we performed a long-term systemic treatment using rGDF5 and showed its effectiveness in counteracting age-related muscle wasting, improving muscle function (17.8% of absolute maximal force increase, P = 0.0079), ensuring neuromuscular connectivity and preventing neuromuscular junction degeneration (7.96% of AchR area increase, P = 0.0125). In addition, in human muscle biopsies, we found the same age-related alterations than those observed in mice and improved by GDF5 and reproduced its major effects on human cells, suggesting this treatment as efficient in humans. Overall, these data provide a foundation to examine the curative potential of GDF5 drug in clinical trials for sarcopenia and, eventually, other neuromuscular diseases.

Mechanism of post-tetanic depression of slow muscle fibres.

A brief tetanic stimulation has a very different effect on the subsequent isometric twitch force of fast and slow skeletal muscles. Fast muscle responds with an enhanced twitch force which doubles that of the pre-tetanic value, whereas slow muscle depresses the post-tetanic twitch by about 20%. Twitch potentiation of fast muscle has long been known to be due to myosin light chain 2 phosphorylation. It is proposed that post-tetanic twitch depression in slow muscle is due to the dephosphorylation of the slow isoform of the thick filament protein, myosin-binding protein-C, by Ca2+/calmodulin-activated phosphatase calcineurin, whilst its phosphorylation underlies the force enhancement due to β-adrenergic stimulation in slow and fast muscle.

Fermented Yak-Kong using Bifidobacterium animalis derived from Korean infant intestine effectively relieves muscle atrophy in an aging mouse model.

Yak-Kong (YK) is a small black soybean widely cultivated in Korea. It is considered to have excellent health functionality, as it has been reported to have better antioxidant efficacy than conventional black or yellow soybeans. Since YK has been described as good for the muscle health of the elderly in old oriental medicine books, this study sought to investigate the effect of fermented YK with Bifidobacterium animalis subsp. lactis LDTM 8102 (FYK) on muscle atrophy. In C2C12 mouse myoblasts, FYK elevated the expression of MyoD, total MHC, phosphorylated AKT, and PGC1α. In addition, two kinds of in vivo studies were conducted using both an induced and normal aging mouse model. The behavioral test results showed that in the induced aging mouse model, FYK intake alleviated age-related muscle weakness and loss of exercise performance. In addition, FYK alleviated muscle mass decrease and improved the expression of biomarkers including total MHC, myf6, phosphorylated AKT, PGC1α, and Tfam, which are related to myoblast differentiation, muscle protein synthesis, and mitochondrial generation in the muscle. In the normal aging model, FYK consumption did not increase muscle mass, but did upregulate the expression levels of biomarkers related to myoblast differentiation, muscle hypertrophy, and muscle function. Furthermore, it mitigated age-related declines in skeletal muscle force production and functional limitation by enhancing exercise performance and grip strength. Taken together, the results suggest that FYK has the potential to be a new functional food material that can alleviate the loss of muscle mass and strength caused by aging and prevent sarcopenia.

Decoding the forces that shape muscle stem cell function.

Skeletal muscle is a force-producing organ composed of muscle tissues, connective tissues, blood vessels, and nerves, all working in synergy to enable movement and provide support to the body. While robust biomechanical descriptions of skeletal muscle force production at the body or tissue level exist, little is known about force application on microstructures within the muscles, such as cells. Among various cell types, skeletal muscle stem cells reside in the muscle tissue environment and play a crucial role in driving the self-repair process when muscle damage occurs. Early evidence indicates that the fate and function of skeletal muscle stem cells are controlled by both biophysical and biochemical factors in their microenvironments, but much remains to accomplish in quantitatively describing the biophysical muscle stem cell microenvironment. This book chapter aims to review current knowledge on the influence of biophysical stresses and landscape properties on muscle stem cells in heath, aging, and diseases.

A low-cost 2-D sarcomere model to demonstrate titin-related mechanisms for force production.

Given the recently proposed three-filament theory of muscle contraction, we present a low-cost physical sarcomere model aimed at illustrating the role of titin in the production of active force in skeletal muscle. With inexpensive materials, it is possible to illustrate actin-myosin cross-bridge interactions between the thick and thin filaments and demonstrate the two different mechanisms by which titin is thought to contribute to active and passive muscle force. Specifically, the model illustrates how titin, a molecule with springlike properties, may increase its stiffness by binding free calcium upon muscle activation and reducing its extensible length by attaching itself to actin, resulting in the greater force-generating capacity after an active than a passive elongation that has been observed experimentally. The model is simple to build and manipulate, and demonstration to high school students was shown to result in positive perception and improved understanding of the otherwise complex titin-related mechanisms of force production in skeletal and cardiac muscles.NEW & NOTEWORTHY Our physical sarcomere model illustrates not only the classic view of muscle contraction, the sliding filament and cross-bridge theories, but also the newly discovered role of titin in force regulation, called the three-filament theory. The model allows for easy visualization of the role of titin in muscle contraction and aids in explaining complex muscle properties that are not captured by the traditional cross-bridge theory.

Skeletal muscle-specific inducible AMPKα1/α2 knockout mice develop muscle weakness, glycogen depletion, and fibrosis that persists during disuse atrophy.

The 5' adenosine monophosphate-activated protein kinase (AMPK) is an important skeletal muscle regulator implicated as a possible therapeutic target to ameliorate the local undesired deconditioning of disuse atrophy. However, the muscle-specific role of AMPK in regulating muscle function, fibrosis, and transcriptional reprogramming during physical disuse is unknown. The purpose of this study was to determine how the absence of both catalytic subunits of AMPK in skeletal muscle influences muscle force production, collagen deposition, and the transcriptional landscape. We generated skeletal muscle-specific tamoxifen-inducible AMPKα1/α2 knockout (AMPKα-/-) mice that underwent 14 days of hindlimb unloading (HU) or remained ambulatory for 14 days (AMB). We found that AMPKα-/- during ambulatory conditions altered body weight and myofiber size, decreased muscle function, depleted glycogen stores and TBC1 domain family member 1 (TBC1D1) phosphorylation, increased collagen deposition, and altered transcriptional pathways. Primarily, pathways related to cellular senescence and mitochondrial biogenesis and function were influenced by the absence of AMPKα. The effects of AMPKα-/- persisted, but were not worsened, following hindlimb unloading. Together, we report that AMPKα is necessary to maintain skeletal muscle quality.NEW & NOTEWORTHY We determined that skeletal muscle-specific AMPKα knockout (KO) mice display functional, fibrotic, and transcriptional alterations before and during muscle disuse atrophy. We also observed that AMPKα KO drives muscle fibrosis and pathways related to cellular senescence that continues during the hindlimb unloading period.

The potassium-glycogen interaction on force and excitability in mouse skeletal muscle: implications for fatigue.

A reduced muscle glycogen content and potassium (K+ ) disturbances across muscle membranes occur concomitantly during repeated intense exercise and together may contribute to skeletal muscle fatigue. Therefore, we examined whether raised extracellular K+ concentration ([K+ ]o ) (4 to 11 mM) interacts with lowered glycogen to reduce force production. Isometric contractions were evoked in isolated mouse soleus muscles (37°C) using direct supramaximal field stimulation. (1) Glycogen declined markedly in non-fatigued muscle with >2h exposure in glucose-free physiological saline compared with control solutions (11 mM glucose), i.e. to <45% control. (2) Severe glycogen depletion was associated with increased 5'-AMP-activated protein kinase activity, indicative of metabolic stress. (3) The decline of peak tetanic force at 11mM [K+ ]o was exacerbated from 67% initial at normal glycogen to 22% initial at lowered glycogen. This was due to a higher percentage of inexcitable fibres (71% vs. 43%), yet without greater sarcolemmal depolarisation or smaller amplitude action potentials. (4) Returning glucose while at 11mM [K+ ]o increased both glycogen and force. (5) Exposure to 4mM [K+ ]o glucose-free solutions (15 min) did not increase fatiguability during repeated tetani; however, after recovery there was a greater force decline at 11mM [K+ ]o at lower than normal glycogen. (6) An important exponential relationship was established between relative peak tetanic force at 11mM [K+ ]o and muscle glycogen content. These findings provide direct evidence of a synergistic interaction between raised [K+ ]o and lowered muscle glycogen as the latter shifts the peak tetanic force-resting EM relationship towards more negative resting EM due to lowered sarcolemmal excitability, which hence may contribute to muscle fatigue. KEY POINTS: Diminished muscle glycogen levels and raised extracellular potassium concentrations ([K+ ]o ) occur simultaneously during intense exercise and together may contribute to muscle fatigue. Prolonged exposure of isolated non-fatigued soleus muscles of mice to glucose-free physiological saline solutions markedly lowered muscle glycogen levels, as does fatigue then recovery in glucose-free solutions. For both approaches, the subsequent decline of maximal force at 11mM[K+ ]o , which mimics interstitial [K+ ] levels during intense exercise, was exacerbated at lowered compared with normal glycogen. This was mainly due to many more muscle fibres becoming inexcitable. We established an important relationship that provides evidence of a synergistic interaction between raised [K+ ]o and lowered glycogen content to reduce force production. This paper indicates that partially lowered muscle glycogen (and/or metabolic stress) together with elevated interstitial [K+ ] interactively lowers muscle force, and hence may diminish performance especially during repeated high-intensity exercise.

Abstract 18258: Leucine Supplementation Improves Myocardial Function in a HFpEF Rat Model

Introduction: We previously documented that leucine supplementation protects from skeletal muscle atrophy by increasing skeletal muscle mass and force during hind limb immobilization. One of the proposed underlying mechanisms was the suppression of HDAC4 protein expression and increased nuclei accretion. Hypothesis: The present study aimed to examine whether the protective effects of leucine supplementation are also observed in the myocardium in an animal model of heart failure with preserved ejection fraction (HFpEF) and to assess the role of HDAC4 modulation. Methods: Female ZSF1 lean (control, con) and obese (HFpEF) rats were randomized at an age of 20 weeks to one of three groups: lean rats receiving standard chow (con), obese rats receiving standard chow (HFpEF), and obese rats receiving leucine-supplemented chow (3% w/w) for 12 weeks (HFpEF/leu). All animals underwent echocardiography at baseline and after 6 and 12 weeks. Prior to organ harvest left ventricular (LV) hemodynamics were measured invasively, blood samples were taken, cardiac mitochondrial function was assessed and LV tissue was collected for molecular and histological analyses. Results: Assessment of diastolic function revealed a 20% reduction of E/é in the HFpEF/leu group compared to untreated HFpEF. While no alterations of left ventricular hemodynamics were observed between the groups, stiffness factor β was significantly reduced in leucine-treated rats. Left ventricular mRNA levels of Collagen type I a1 and matrix metalloproteinase 2 were reduced by 25% and 22%, respectively. Assessment of mitochondrial oxidative phosphorylation revealed a significant improvement of complex-I- and complex-II mediated respiration in leucine-treated rats compared to HFpEF. Finally, HDAC4 protein expression levels were reduced by 40% in the HFpEF/leu group compared to HFpEF. Conclusions: This study presents beneficial effects of leucine supplementation on cardiac function, remodeling, and metabolic parameters in an HFpEF rat model. These benefits may be attributed to suppressed HDAC4 expression.

Automatic extraction and measurement of ultrasonic muscle morphological parameters based on multi-stage fusion and segmentation

BackgroundEstimating skeletal muscle force output and structure requires measurement of morphological parameters including muscle thickness, pennation angle, and fascicle length. The identification of aponeurosis and muscle fascicles from medical images is required to measure these parameters accurately. MethodsThis paper introduces a multi-stage fusion and segmentation model (named MSF-Net), to precisely extract muscle aponeurosis and fascicles from ultrasound images. The segmentation process is divided into three stages of feature fusion modules. A prior feature fusion module (PFFM) is designed in the first stage to fuse prior features, thus enabling the network to focus on the region of interest and eliminate image noise. The second stage involves the addition of multi-scale feature fusion module (MS-FFM) for effective fusion of elemental information gathered from different scales. This process enables the precise extraction of muscle fascicles of varied sizes. Finally, the high-low-level feature fusion attention module (H-LFFAM) is created in the third stage to selectively reinforce features containing useful information. ResultsOur proposed MSF-Net outperforms other methods and achieves the highest evaluation metrics. In addition, MSF-Net can obtain similar results to manual measurements by clinical experts. The mean deviation of muscle thickness and fascicle length was 0.18 mm and 1.71 mm, and the mean deviation of pennation angle was 0.31°. ConclusionsMSF-Net can accurately extract muscle morphological parameters, which enables medical experts to evaluate muscle morphology and function, and guide rehabilitation training. Therefore, MSF-Net provides a complementary imaging tool for clinical assessment of muscle structure and function.

INVESTIGATION OF THE LOADING AT THE KNEE JOINT COMPLEX USING AN EMG-BASED CONSTITUTIVE LAW FOR SKELETAL MUSCLE FORCE

This study investigated the predictive ability of the skeletal muscle force model presented by Knodel et al. [Knodel NB, Lawson LB, Nauman EA, “An emg-based constitutive law for force generation in skeletal muscle-part i: Model development,” J Biomech Eng (in press), doi: 10.1115/1.4053568] on the knee joint. It has previously been validated on the ankle joint [Knodel NB, Calvert LB, Bywater EA, Lamia JP, Patel SN, Nauman EA, “An emg-based constitutive law for force generation in skeletal muscle-part ii: Model validation on the ankle joint complex,” Submitted for Publication] and this paper aimed to identify how well it, and the solution process, performed on a more complex articulation. The knee joint’s surrounding musculoskeletal tissue loading was also identified. Ten subjects (five male and five female) performed six exercises targeting the muscles that cross the knee joint. Motion capture, electromyography, and force plate data was collected during the exercises for use in the analysis program written in MATLAB and magnetic resonance images were used to observe subject-specific ligament and tendon data at the knee articulation. OpenSim [Delp, SL, Anderson FC, Arnold AS, Loan P, Habib A, John CT, Guendelman E, Thelen DG, “Opensim: Open-source software to create and analyze dynamic simulations of movement,” IEEE Trans Biomed Eng 54(11):1940–1950, 2007, doi: 10.1109/TBME.2007.901024] was used for scaling a generic lower extremity anatomical model of each subject. Five of the six exercises were used to calculate each muscle’s constant, [Formula: see text] [Knodel NB, Lawson LB, Nauman EA, “An emg-based constitutive law for force generation in skeletal muscle-part i: Model development,” J Biomech Eng (in press), doi: 10.1115/1.4053568; Knodel NB, Calvert LB, Bywater EA, Lamia JP, Patel SN, Nauman EA, “An emg-based constitutive law for force generation in skeletal muscle-part ii: Model validation on the ankle joint complex,” Submitted for Publication], and the sixth was used as a testing set to identify the model’s predictive ability. Average percent errors ranged from 9.4% to 26.5% and the average across all subjects was 20.6%. The solution process produced physiologically relevant muscle forces and the surrounding tissue loading behaved as expected between the various exercises without approaching respective tensile strength values.

Mimicking the Tendon Microenvironment to Enhance Skeletal Muscle Adhesion and Longevity in a Functional Microcantilever Platform.

Microcantilever platforms are functional models for studying skeletal muscle force dynamics in vitro. However, the contractile force generated by the myotubes can cause them to detach from the cantilevers, especially during long-term experiments, thus impeding the chronic investigations of skeletal muscles for drug efficacy and toxicity. To improve the integration of myotubes with microcantilevers, we drew inspiration from the elastomeric proteins, elastin and resilin, that are present in the animal and insect worlds, respectively. The spring action of these proteins plays a critical role in force dampening in vivo. In animals, elastin is present in the collagenous matrix of the tendon which is the attachment point of muscles to bones. The tendon microenvironment consists of elastin, collagen, and an aqueous jelly-like mass of proteoglycans. In an attempt to mimic this tendon microenvironment, elastin, collagen, heparan sulfate proteoglycan, and hyaluronic acid were deposited on a positively charged silane substrate. This enabled the long-term survival of mechanically active myotubes on glass and silicon microcantilevers for over 28 days. The skeletal muscle cultures were derived from both primary and induced pluripotent stem cell (iPSC)-derived human skeletal muscles. Both types of myoblasts formed myotubes which survived for five weeks. Primary skeletal muscles and iPSC-derived skeletal muscles also showed a similar trend in fatigue index values. Upon integration with the microcantilever system, the primary muscle and iPSC-derived myotubes were tested successively over a one month period, thus paving the way for long-term chronic experiments on these systems for both drug efficacy and toxicity studies.

N-acetylneuraminate pyruvate lyase controls sialylation of muscle glycoproteins essential for muscle regeneration and function.

Deleterious variants in N-acetylneuraminate pyruvate lyase (NPL) cause skeletal myopathy and cardiac edema in humans and zebrafish, but its physiological role remains unknown. We report generation of mouse models of the disease: NplR63C, carrying the human p.Arg63Cys variant, and Npldel116 with a 116-bp exonic deletion. In both strains, NPL deficiency causes drastic increase in free sialic acid levels, reduction of skeletal muscle force and endurance, slower healing and smaller size of newly formed myofibers after cardiotoxin-induced muscle injury, increased glycolysis, partially impaired mitochondrial function, and aberrant sialylation of dystroglycan and mitochondrial LRP130 protein. NPL-catalyzed degradation of sialic acid in the muscle increases after fasting and injury and in human patient and mouse models with genetic muscle dystrophy, demonstrating that NPL is essential for muscle function and regeneration and serves as a general marker of muscle damage. Oral administration of N-acetylmannosamine rescues skeletal myopathy, as well as mitochondrial and structural abnormalities in NplR63C mice, suggesting a potential treatment for human patients.

An electrical stimulation intervention protocol to prevent disuse atrophy and muscle strength decline: an experimental study in rat

BackgroundSkeletal muscle is negatively impacted by conditions such as spaceflight or prolonged bed rest, resulting in a dramatic decline in muscle mass, maximum contractile force, and muscular endurance. Electrical stimulation (ES) is an essential tool in neurophysiotherapy and an effective means of preventing skeletal muscle atrophy and dysfunction. Historically, ES treatment protocols have used either low or high frequency electrical stimulation (LFES/HFES). However, our study tests the use of a combination of different frequencies in a single electrical stimulation intervention in order to determine a more effective protocol for improving both skeletal muscle strength and endurance.MethodsAn adult male SD rat model of muscle atrophy was established through 4 weeks of tail suspension (TS). To investigate the effects of different frequency combinations, the experimental animals were treated with low (20 Hz) or high (100 Hz) frequency before TS for 6 weeks, and during TS for 4weeks. The maximum contraction force and fatigue resistance of skeletal muscle were then assessed before the animals were sacrificed. The muscle mass, fiber cross-sectional area (CSA), fiber type and related protein expression were examined and analyzed to gain insights into the mechanisms by which the ES intervention protocol used in this study regulates muscle strength and endurance.ResultsAfter 4 weeks of unloading, the soleus muscle mass and fiber CSA decreased by 39% and 58% respectively, while the number of glycolytic muscle fibers increased by 21%. The gastrocnemius muscle fibers showed a 51% decrease in CSA, with a 44% decrease in single contractility and a 39% decrease in fatigue resistance. The number of glycolytic muscle fibers in the gastrocnemius also increased by 29%. However, the application of HFES either prior to or during unloading showed an improvement in muscle mass, fiber CSA, and oxidative muscle fibers. In the pre-unloading group, the soleus muscle mass increased by 62%, while the number of oxidative muscle fibers increased by 18%. In the during unloading group, the soleus muscle mass increased by 29% and the number of oxidative muscle fibers increased by 15%. In the gastrocnemius, the pre-unloading group showed a 38% increase in single contractile force and a 19% increase in fatigue resistance, while in the during unloading group, a 21% increase in single contractile force and a 29% increase in fatigue resistance was observed, along with a 37% and 26% increase in the number of oxidative muscle fibers, respectively. The combination of HFES before unloading and LFES during unloading resulted in a significant elevation of the soleus mass by 49% and CSA by 90%, with a 40% increase in the number of oxidative muscle fibers in the gastrocnemius. This combination also resulted in a 66% increase in single contractility and a 38% increase in fatigue resistance.ConclusionOur results indicated that using HFES before unloading can reduce the harmful effects of muscle unloading on the soleus and gastrocnemius muscles. Furthermore, we found that combining HFES before unloading with LFES during unloading was more effective in preventing muscle atrophy in the soleus and preserving the contractile function of the gastrocnemius muscle.

Sex differences in isolated skeletal muscle force production: a spectrum of responses

Whether sex differences in skeletal muscle microvascular responses differ in response to muscle contraction is currently unknown. To determine vascular responses to muscle contraction in vivo we use intravital microscopy using thin skeletal muscle models of mixed fibre types (i.e. hamster retractor muscle). However, to effectively quantify this relationship in vivo, the initial muscle contraction stimulus must be constant across the sexes and thus, we must first determine whether there are sex differences in skeletal muscle force generation. We hypothesized that sexual dimorphisms in force production would not be apparent in muscles of any fibre type composition. To test this hypothesis, we used skeletal muscles with unique fibre type characteristics and oxidative capacities. Mouse soleus (SOL: primarily type 1 &gt;75%), extensor digitorum longus (EDL: primarily type IIB ~60%), and costal diaphragm (DIA: primarily type IIA ~55%) muscles were isolated from adult male and female CD-1 mice. The retractor muscle (RET: primarily type IIX ~58%) was also isolated from adult male and female Syrian hamsters. Muscles were electrically stimulated in vitro and force production was quantified at optimal length during a force-frequency test (1-120Hz) and twitch (1Hz) contractions. In mice, females generated greater force during the force-frequency test in SOL (10-120Hz) and EDL (30-120Hz) and generated greater twitch forces compared to males (SOL: female 34.7 ± 2.4 mN/mm 2 , n=21 vs male 27.8 ± 2.0 mN/mm 2 , n=17; EDL: female 41.2 ± 2.2 mN/mm 2 , n=35 vs male 32.5 ± 2.1 mN/mm 2 , n=29). Females generated greater force during the force frequency test in DIA at 30Hz only and no significant sex differences were observed in DIA twitch force (female 0.24 ± 0.02 g/mg, n=19 vs male 0.20 ± 0.02 g/mg, n=21). In hamster RET no significant sex differences in force were observed during the force frequency test or during the twitch contraction (female 0.13 ± 0.01 g/mg, n=19 vs male 0.14 ± 0.02 g/mg, n=20). Collectively, these findings demonstrate that sex differences in force production were observed in muscles primarily comprised of slow oxidative (type 1) and fast glycolytic (type IIB) fibres but not in muscles primarily comprised of intermediate fibres (type IIA and type IIX). These data show that on a spectrum of muscle fibres from slow oxidative to fast glycolytic, sex differences are most apparent on the extremes of the spectrum, whereas sexual dimorphisms in the middle of the spectrum are not apparent. This work is supported by NSERC. NSERC This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.