Vastus Lateralis Muscle Fibers Research Articles

Melatonin induces fiber switching by improvement of mitochondrial oxidative capacity and function via NRF2/RCAN/MEF2 in the vastus lateralis muscle from both sex Zücker diabetic fatty rats.

The positive role of melatonin in obesity control and skeletal muscle (SKM) preservation is well known. We recently showed that melatonin improves vastus lateralis muscle (VL) fiber oxidative phenotype. However, fiber type characterization, mitochondrial function, and molecular mechanisms that underlie VL fiber switching by melatonin are still undefined. Our study aims to investigate whether melatonin induces fiber switching by NRF2/RCAN/MEF2 pathway activation and mitochondrial oxidative metabolism modulation in the VL of both sex Zücker diabetic fatty (ZDF) rats. 5-weeks-old male and female ZDF rats (N=16) and their age-matched lean littermates (ZL) were subdivided into two subgroups: control (C) and orally treated with melatonin (M) (10 mg/kg/day) for 12 weeks. Interestingly, melatonin increased oxidative fibers amounts (Types I and IIa) counteracting the decreased levels found in the VL of obese-diabetic rats, and upregulated NRF2, calcineurin and MEF2 expression. Melatonin also restored the mitochondrial oxidative capacity increasing the respiratory control ratio (RCR) in both sex and phenotype rats through the reduction of the proton leak component of respiration (state 4). Melatonin also improved the VL mitochondrial phosphorylation coefficient and modulated the total oxygen consumption by enhancing complex I, III and IV activity, and fatty acid oxidation (FAO) in both sex obese-diabetic rats, decreasing in male and increasing in female the complex II oxygen consumption. These findings suggest that melatonin treatment induces fiber switching in SKM improving mitochondrial functionality by NRF2/RCAN/MEF2 pathway activation.

Expression of intramuscular extracellular matrix proteins in vastus lateralis muscle fibres between atrophic and non-atrophic COPD.

Extracellular matrix (ECM) proteins are the major constituents of the muscle cell micro-environment, imparting instructive signalling, steering cell behaviour and controlling muscle regeneration. ECM remodelling is among the most affected signalling pathways in COPD and aged muscle. As a fraction of COPD patients present muscle atrophy, we questioned whether ECM composition would be altered in patients with peripheral muscle wasting (atrophic COPD) compared to those without muscle wasting (non-atrophic COPD). A set of ECM molecules with known impact on myogenesis were quantified in vastus lateralis muscle biopsies from 29 COPD patients (forced expiratory volume in 1 s 55±12% predicted) using ELISA and real-time PCR. COPD patients were grouped to atrophic or non-atrophic based on fat-free mass index (<17 or ≥17 kg·m-2). Atrophic COPD patients presented a lower average vastus lateralis muscle fibre cross-sectional area (3872±258 μm2) compared to non-atrophic COPD (4509±198 μm2). Gene expression of ECM molecules was found significantly lower in atrophic COPD compared to non-atrophic COPD for collagen type I alpha 1 chain (COL1A1), fibronectin (FN1), tenascin C (TNC) and biglycan (BGN). In terms of protein levels, there were no significant differences between the two COPD cohorts for any of the ECM molecules tested. Although atrophic COPD presented decreased contractile muscle tissue, the differences in ECM mRNA expression between atrophic and non-atrophic COPD were not translated at the protein level, potentially indicating an accumulation of long-lived ECM proteins and dysregulated proteostasis, as is typically observed during deconditioning and ageing.

Sex Differences in Quadriceps Atrophy After Anterior Cruciate Ligament Tear.

Female athletes lag behind their male counterparts in recovery from anterior cruciate ligament (ACL) injury. Quadriceps muscle size and strength are crucial factors for regaining function after ACL injury, but little is known about how these metrics vary due to biological sex. Female patients have reduced vastus lateralis fiber cross-sectional area (CSA) and lower quadriceps strength after ACL injury than male patients. Cross-sectional study. Level 4. A total of 60 participants with recent ACL tear were evaluated for vastus lateralis muscle fiber CSA, isometric quadriceps peak torque, and quadriceps rate of torque development. Linear mixed models were fit to determine differences across sex and limb for each variable of interest. The female group averaged almost 20% atrophy between limbs (P < 0.01), while the male group averaged just under 4% (P = 0.05). Strength deficits between limbs were comparable between female and male groups. Immediately after ACL injury, female patients have greater between-limb differences in muscle fiber CSA but between-limb strength deficits comparable with those of male patients. These results indicate that the underpinnings of strength loss differ based on biological sex, and thus individual patients could benefit from a sex-specific treatment approach to ACL injury.

Immunohistochemical analysis of the quadriceps femoris muscle before and after total knee arthroplasty

BackgroundThe number of total knee arthroplasties (TKA) has increased steadily with the aging of the population. This surgical procedure is recognized for its success in pain relief and restoration of knee function. However, decreased quadriceps femoris (QF) muscle strength after TKA is frequently observed but with unknown etiology. Evidence suggests that the location of the operative incision (i.e., surgical access) can influence QF muscle structure and function. The present study aimed to assess the fiber type composition, structure and assembly of the QF's vastus medialis (VM) and vastus lateralis (VL) muscles before and after TKA. MethodsMuscle biopsies (VM and VL muscles) were collected from patients previously submitted to TKA via the medial parapatellar route and undergoing TKA revision (main group, n = 9) and patients with osteoarthrosis (OA) who were due to undergo TKA (control group: n = 18). The biopsied muscle tissue was prepared, stored, and then sectioned in a cryostat at −25 °C. The tissue sections were evaluated using routine staining techniques in pathological anatomy and histochemistry. ResultsThe normal mosaic pattern of the medial and lateral knee muscles was observed in the main and control groups, with no evidence of peripheral nerve damage. Notably, 88.9 % of the patients exhibited mild to severe VL atrophy, while only 11.1 % of patients in the control group presented this feature (P < 0.001). ConclusionsThe medial parapatellar incision for TKA surgical access does not generate definitive morphological changes in the VM and VL muscle fibers but may contribute to VL atrophy.

Development of skeletal muscle atrophy and intermuscular adipose tissue in patients with early breast cancer treated with chemotherapy.

Chemotherapy is a common therapy to treat patients with breast cancer but also leads to skeletal muscle deconditioning. Skeletal muscle deconditioning is multifactorial and intermuscular adipose tissue (IMAT) accumulation is closely linked to muscle dysfunction. To date, there is no clinical study available investigating IMAT development through a longitudinal protocol and the underlying mechanisms remain unknown. Our study was dedicated to investigating IMAT content in patients with early breast cancer who were treated with chemotherapy and exploring the subsequent cellular mechanisms involved in its development. We included 13 women undergoing chemotherapy. Muscle biopsies and ultrasonography assessment were performed before and after chemotherapy completion. Histological and Western blotting analyses were conducted. We found a substantial increase in protein levels of three mature adipocyte markers (perilipin, +901%; adiponectin, +135%; FABP4, +321%; P < 0.05). These results were supported by an increase in oil red O-positive staining (+358%; P < 0.05). A substantial increase in PDGFRα protein levels was observed (+476%; P < 0.05) highlighting an increase in fibro-adipogenic progenitors (FAPs) content. The cross-sectional area of the vastus lateralis muscle fibers substantially decreased (-21%; P < 0.01), and muscle architecture was altered, as shown by a decrease in fascicle length (-15%; P < 0.05) and a decreasing trend in muscle thickness (-8%; P = 0.08). We demonstrated both IMAT development and muscle atrophy in patients with breast cancer who were treated with chemotherapy. FAPs, critical stem cells inducing both IMAT development and skeletal muscle atrophy, also increased, suggesting that FAPs likely play a critical role in the skeletal muscle deconditioning observed in patients with breast cancer who were treated with chemotherapy.

Active behavior of skeletal muscle and modeling sarcopenia with a mixture active strain approach based constitutive model

Different continuum hyperelastic constitutive models for the contraction of the skeletal muscle exist in literature. In this work, we compare the models developed by Ehret et al (2011) and Riccobelli and Ambrosi (2019). We follow the mixture active strain approach of the latter but employ the strain energy density functions proposed by the former for modeling the contraction. The model excellently recovers the experimental stress–stretch data of a passive and activated rat tibialis anterior muscle. The model is further improved by introducing a decay deformation tensor into the multiplicative decomposition of the deformation gradient, which helps model the geriatric syndrome called sarcopenia. This improved model accurately reproduces the experimental stress–stretch data of passive and activated human type II vastus lateralis muscle fibers of both younger and older adults and suggests that changes in all muscle parameters are responsible for explaining sarcopenia.

Weak Association Between Vastus Lateralis Muscle Fiber Composition and Fascicle Length in Young Untrained Females.

The aim of the study was to investigate the relationships between vastus lateralis muscle fiber length and fiber type composition in individuals with minimal exposure to systematic resistance/power training. In sixty female physical education students (age: 21.03 ± 2.1 years, body weight: 59.8 ± 9.7 kg, body height: 166.2 ± 6.5 cm), with no experience in systematic training, lean body mass, VL muscle architecture and fiber composition type, countermovement jumping (CMJ) performance, and isometric leg press rate of force development were evaluated. Data were analyzed for all participants, as well as two equally numbered groups assigned according to their maximum countermovement jumping power (High-Power or Low-Power group). Significant but low correlations were found between type II muscle fiber percentage and fascicle length (N = 60, p < 0.05). Significant correlations were found between type IIa and IIx muscle fiber percentage cross-sectional area (%CSA) and fascicle length (N = 60; r = 0.321, and r = 0.378; respectively, p < 0.05). These correlations were higher for the High-Power group (r = 0.499, and r = 0.522; respectively, p < 0.05), and lower, and nonsignificant, for the Low-Power group. The best predictor of strength/power performance was the lean body mass of the lower extremities (r = 0.389–0.645, p < 0.05). These results suggest that in females with minimal exposure to systematic training, fascicle length may be weakly linked with type II fiber areas, only in females with high-power profiles.

Effects of NMES pulse width and intensity on muscle mechanical output and oxygen extraction in able-bodied and paraplegic individuals.

Neuromuscular Electrical Stimulation (NMES) is commonly used in neuromuscular rehabilitation protocols, and its parameters selection substantially affects the characteristics of muscle activation. Here, we investigated the effects of short pulse width (200µs) and higher intensity (short-high) NMES or long pulse width (1000µs) and lower intensity (long-low) NMES on muscle mechanical output and fractional oxygen extraction. Muscle contractions were elicited with 100Hz stimulation frequency, and the initial torque output was matched by adjusting stimulation intensity. Fourteen able-bodied and six spinal cord-injured (SCI) individuals participated in the study. The NMES protocol (75 isometric contractions, 1-s on-3-s off) targeting the knee extensors was performed with long-low or short-high NMES applied over the midline between anterior superior iliac spine and patella protrusion in two different days. Muscle work was estimated by torque-time integral, contractile properties by rate of torque development and half-relaxation time, and vastus lateralis fractional oxygen extraction was assessed by Near-Infrared Spectroscopy (NIRS). Torque-time integral elicited by the two NMES paradigms was similar throughout the stimulation protocol, with differences ranging between 1.4% (p = 0.877; able-bodied, mid-part of the protocol) and 9.9% (p = 0.147; SCI, mid-part of the protocol). Contractile properties were also comparable in the two NMES paradigms. However, long-low NMES resulted in higher fractional oxygen extraction in able-bodied (+ 36%; p = 0.006). Long-low and short-high NMES recruited quadriceps femoris motor units that demonstrated similar contractile and fatigability properties. However, long-low NMES conceivably resulted in the preferential recruitment of vastus lateralis muscle fibers as detected by NIRS.

Association of muscle fiber composition with health and exercise-related traits in athletes and untrained subjects.

Skeletal muscle is a heterogenous and metabolically active tissue, the composition of which is associated with multiple traits. The aim of the study was to determine whether there are additional health and exercise-related traits associated with muscle fiber composition in athletes and non-athletes. This study recruited 164 Russian participants (51 endurance and 48 power athletes; 65 controls). Vastus lateralis muscle fiber composition was assessed by immunohistochemistry. Slow-twitch muscle fiber percentage (STMF%) was significantly greater in endurance than power athletes and non-athletes, and in non-athlete females than males. STMF% was positively associated with athletes’ training frequency, non-athletes’ and endurance athletes’ age, endurance athletes’ competition level and chest depth, and power athletes’ training age. STMF% was negatively associated with diastolic blood pressure in power athletes and with systolic blood pressure and reaction time in non-athletes. In all participants, STMF% was positively associated with age, tolerance to long distance exercise, chest depth and fracture incidence, and negatively with systolic blood pressure and resting heart rate. Age, sex and training frequency explained 10.6% and 13.2% of the variance in STMF% in endurance and power athletes, respectively. This is one of the most comprehensive studies involving athletes and untrained subjects and provides novel information concerning associations of increased STMF percentage with lower resting heart rate, better tolerance to long distances, faster reaction time and larger chest depth. On the other hand, the increased percentage of fast-twitch muscle fibers was associated with rare fracture incidence.

Human skeletal muscle fiber type percentage and area after reduced muscle use: A systematic review and meta-analysis.

The main objective of this systematic review was to examine the effect of reduced muscle activity on the relative number of type 1 muscle fibers (%) in the human vastus lateralis muscle. Other objectives were changes in type 2A and 2X percentages and muscle fiber cross-sectional area. We conducted systematic literature searches in eight databases and included studies assessing type 1 fiber percentage visualized by ATPase or immunohistochemical staining before and after a period (≥14days) of reduced muscle activity. The reduced muscle activity models were detraining, leg unloading, and bed rest. Forty-two studies comprising 451 participants were included. Effect sizes were calculated as the mean difference between baseline and follow-up and Generic Inverse Variance tests with random-effects models were used for the weighted summary effect size. Overall, the mean type 1 muscle fiber percentage was significantly reduced after interventions (-1.94%-points, 95% CI [-3.37, -0.51], P=.008), with no significant differences between intervention models (P=.86). Meta-regression showed no effect of study duration on type 1 fiber percentage (P=.98). Conversely, the overall type 2X fiber percentage increased after reduced muscle activity (P<.001). The CSA of the muscle fiber types decreased after the study period (all P-values<0.001) with greater reductions in type 2 than type 1 fibers (P<.001). The result of this meta-analysis display that the type 1 muscle fiber percentage decrease as a result of reduced muscle activity, although the effect size is relatively small.

The influence of functional electrical stimulation on the properties of vastus lateralis fibres following total knee arthroplasty

The influence of functional electrical muscle stimulation (FES) on selected properties of vastus lateralis muscle fibres was studied in patients recovering from total knee arthroplasty for osteoarthritis. Prior to surgery, on the average, muscle biopsies from the vastus lateralis could be characterized as having a predominance of Type I fibres which were significantly larger in cross-sectional area than the Type II fibres in the same sample. Following surgery, muscle biopsies from a group of patients (n = 7) which received continuous passive motion and no FES, exhibited a marked increase in the proportion of Type II fibres along with a general atrophy of both the Type I and Type II fibres. Patients receiving passive motion and FES (n = 9) also showed an increase in the relative percentage of Type II fibres. Post-operatively, however, there was no significant reduction in fibre area in the stimulated muscles. These data suggest that FES was effective in attenuating the muscle atrophy associated with total knee arthroplasty but had no influence on those metabolic properties which were related to muscle fibre type classification criteria.

Effects of high-intensity intermittent exercise on the contractile properties of human type I and type II skeletal muscle fibers.

In vitro studies have shown that alterations in redox state can cause a range of opposing effects on the properties of the contractile apparatus in skeletal muscle fibers. To test whether and how redox changes occurring in vivo affect the contractile properties, vastus lateralis muscle fibers from seven healthy young adults were examined at rest (PRE) and following (POST) high-intensity intermittent cycling exercise. Individual mechanically skinned muscle fibers were exposed to heavily buffered solutions at progressively higher free [Ca2+] to determine their force-Ca2+ relationship. Following acute exercise, Ca2+ sensitivity was significantly decreased in type I fibers (by 0.06 pCa unit) but not in type II fibers (0.01 pCa unit). Specific force decreased after the exercise in type II fibers (-18%) but was unchanged in type I fibers. Treatment with the reducing agent dithiothreitol (DTT) caused a small decrease in Ca2+-sensitivity in type II fibers at PRE (by ∼0.014 pCa units) and a significantly larger decrease at POST (∼0.035 pCa units), indicating that the exercise had increased S-glutathionylation of fast troponin I. DTT treatment also increased specific force (by ∼4%), but only at POST. In contrast, DTT treatment had no effect on either parameter in type I fibers at either PRE or POST. In type I fibers, the decreased Ca2+ sensitivity was not due to reversible oxidative changes and may have contributed to a decrease in power production during vigorous exercises. In type II fibers, exercise-induced redox changes help counter the decline in Ca2+-sensitivity while causing a small decline in maximum force.NEW & NOTEWORTHY This study identified important cellular changes occurring in human skeletal muscle fibers following high-intensity intermittent exercise: 1) a decrease in contractile apparatus Ca2+ sensitivity in type I but not type II fibers, 2) a decrease in specific force only in type II muscle fibers, and 3) a redox-dependent increase in Ca2+ sensitivity occurring only in type II fibers, which would help maintain muscle performance by countering the normal metabolite-induced decline in Ca2+ sensitivity.

Muscle Oxygen Supply and Use in Type 1 Diabetes, From Ambient Air to the Mitochondrial Respiratory Chain: Is There a Limiting Step?

Long before clinical complications of type 1 diabetes (T1D) develop, oxygen supply and use can be altered during activities of daily life. We examined in patients with uncomplicated T1D all steps of the oxygen pathway, from the lungs to the mitochondria, using an integrative ex vivo (muscle biopsies) and in vivo (during exercise) approach. We compared 16 adults with T1D with 16 strictly matched healthy control subjects. We assessed lung diffusion capacity for carbon monoxide and nitric oxide, exercise-induced changes in arterial O2 content (SaO2, PaO2, hemoglobin), muscle blood volume, and O2 extraction (via near-infrared spectroscopy). We analyzed blood samples for metabolic and hormonal vasoactive moieties and factors that are able to shift the O2-hemoglobin dissociation curve. Mitochondrial oxidative capacities were assessed in permeabilized vastus lateralis muscle fibers. Lung diffusion capacity and arterial O2 transport were normal in patients with T1D. However, those patients displayed blunted exercise-induced increases in muscle blood volume, despite higher serum insulin, and in O2 extraction, despite higher erythrocyte 2,3-diphosphoglycerate. Although complex I- and complex II-supported mitochondrial respirations were unaltered, complex IV capacity (relative to complex I capacity) was impaired in patients with T1D, and this was even more apparent in those with long-standing diabetes and high HbA1c. [Formula: see text]O2max was lower in patients with T1D than in the control subjects. Early defects in microvascular delivery of blood to skeletal muscle and in complex IV capacity in the mitochondrial respiratory chain may negatively impact aerobic fitness. These findings are clinically relevant considering the main role of skeletal muscle oxidation in whole-body glucose disposal.

Effect of Concurrent Power Training and High-Intensity Interval Cycling on Muscle Morphology and Performance.

Spiliopoulou, P, Zaras, N, Methenitis, S, Papadimas, G, Papadopoulos, C, Bogdanis, GC, and Terzis, G. Effect of concurrent power training and high-intensity interval cycling on muscle morphology and performance. J Strength Cond Res 35(9): 2464-2471, 2021-The aim of the study was to examine the effect of performing high-intensity interval cycling on muscle morphology and performance immediately after power training (PT). Twenty healthy female physical education students were assigned into 2 training groups. One group performed PT, and the other group performed the same PT followed by high-intensity interval aerobic training on a cycle ergometer (PTC). Training was performed 3 days per week for 6 weeks. Countermovement jump (CMJ) height and CMJ power, half-squat maximal strength (1 repetition maximum), maximum aerobic power, vastus lateralis muscle fiber composition, and cross-sectional area (CSA) were evaluated before and after the intervention. Countermovement jump height increased after PT (10.1 ± 6.6%, p = 0.002) but not after PTC (-5.1 ± 10.5%, p = 0.099), with significant difference between groups (p = 0.001). Countermovement jump power increased after PT (4.5 ± 4.9%, p = 0.021) but not after PTC (-2.4 ± 6.4, p = 0.278), with significant difference between groups (p = 0.017). One repetition maximum increased similarly in both groups. Muscle fiber composition was not altered after either PT or PTC. Vastus lateralis muscle fiber CSA increased significantly and similarly after both PT (I: 16.9 ± 16.2%, p = 0.035, ΙΙΑ: 12.7 ± 10.9%, p = 0.008,ΙΙΧ: 15.5 ± 17.1%, p = 0.021) and PTC (Ι: 18.0 ± 23.7%, p = 0.033,ΙΙΑ: 18.2 ± 11.4%, p = 0.001,ΙΙΧ: 25.5 ± 19.6%, p = 0.003). These results suggest that the addition of high-intensity interval cycling to PT inhibits the anticipated increase in jumping performance induced by PT per se. This inhibition is not explained by changes in muscle fiber type composition or vastus lateralis muscle fiber CSA adaptations.

No Difference between Spray Dried Milk and Native Whey Supplementation with Strength Training.

ABSTRACTBackgroundA rapid digestibility and high leucine content are considered important for maximal stimulation of muscle protein synthesis. Consequently, with these properties, native whey may hold greater anabolic potential than milk, when supplemented in combination with strength training. Our aim was to compare the effects of supplementation with milk or native whey, during a 12-wk strength training period, on gains in muscle mass and strength in young adults.MethodsIn this double-blinded, randomized, controlled study a total of 40 untrained young men and women received two daily servings of either milk or native whey containing 20 g of protein, during a 12-wk strength training intervention. Muscle strength, lean mass, thigh muscle cross-sectional area, m. vastus lateralis thickness and muscle fiber cross-sectional area were assessed before and after the training period. In addition, the acute phosphorylation of the anabolic kinases p70S6K, 4E-BP1 and eEF-2 in response to a standardized workout and supplementation was investigated before and after the 12-wk training period.ResultsMuscle mass and strength increased, by all measures applied (5%–16%, P < 0.001), with no differences between groups (P > 0.25). p70S6K phosphorylation increased (~1000%, P < 0.02) 2 h after exercise in the untrained and trained state, but no differences in anabolic signaling were observed between supplements (P > 0.40). No correlation between these acute measures and changes in muscle mass or strength were observed.ConclusionSupplementation with milk or native whey during a 12-wk strength training period did not differentially affect muscle mass and strength in young untrained individuals.

Epigenetic and transcriptomic signatures of human slow‐ and fast‐twitch muscle fibers across the lifespan

The clinical condition of sarcopenia, a hallmark of aging, is characterized by a loss of muscle mass and function, which is more pronounced in fast‐twitch muscle fibers. Exercise can partially mitigate these effects, but little is known about how aging and exercise may impact the control of muscle gene expression through DNA methylation. Therefore, we examined epigenetic and transcriptome signatures of human slow‐twitch (MHC I) and fast‐twitch (MHC IIa) muscle cells (i.e. fibers) in lifelong endurance exercisers (LLE; n=8, 74±1 y, 180±3 cm, 77±2 kg, VO2 max 2.9±0.1 l/min, Quadriceps CSA 68.1±7 cm2), age‐matched healthy non‐exercisers (OH; n=9, 75±1 y, 177±2 cm, 88±3 kg, VO2 max 2.0±0.2 l/min, Quadriceps CSA 55.5±9 cm2), and a young exercising cohort (YE; n=8, 25±1 y, 182±2 cm, 75±3 kg, VO2max 4.1±0.2 l/min, Quadriceps CSA 79.3±9 cm2). For each subject, 96 individual vastus lateralis muscle fibers were separated from a muscle bundle (~5–6 mm length) under a light microscope using fine tweezers. The fiber type (MHC) was determined using SDS‐PAGE before fiber‐type specific DNA and RNA extractions of pooled single muscle fibers (~30 fibers). DNA methylation was assessed using Reduced Representation Bisulfite Sequencing on 32 ng of DNA. Gene expression was examined with RNA‐Sequencing using 9 ng of RNA. Differentially methylated regions (DMRs, methylation difference ≥35%, p&lt;0.05) and differentially expressed transcripts (fold‐change ≥1.5, p&lt;0.01) between fiber types were identified in each subject cohort and overlaid to identify common genes between YE, OH, and LLE. Ingenuity Pathway Analysis software and the gene ontology database were utilized to interpret the biology of the common genes. We identified 183 differentially methylated genes and 141 differentially expressed genes between fiber types, which were common in all three groups (YE, OH, and LLE). The molecular and biological processes of these genes, such as ATP‐dependent microfilament and microtubule motor activities or glycolysis, were consistent with the fiber‐type functional and structural phenotypic differences. We interrogated the 183 differentially methylated genes with the 141 differentially expressed genes, and identified 12 genes with both gene methylation and expression differences between fiber types across all three groups. Seven of them were myosin and troponin genes. These data provide the first fiber‐type specific gene methylation and expression signatures of healthy human skeletal muscle across the lifespan. Future analysis of the epigenetic and transcriptomic signatures specific to YE, LLE, and OH could add knowledge and further our understanding on slow‐ and fast‐twitch fiber biology with aging and exercise. Furthermore, fiber‐type specific epigenetic and transcriptomic characterization of healthy human skeletal muscles will contribute to a better understanding of age‐ and disease‐related muscle dysfunctions.Support or Funding InformationNIH grant R01 AG‐038576 and Ball State University Academic Excellence AwardThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Age-related reduction in single muscle fiber calcium sensitivity is associated with decreased muscle power in men and women

Age-related declines in human skeletal muscle performance may be caused, in part, by decreased responsivity of muscle fibers to calcium (Ca2+). This study examined the contractile properties of single vastus lateralis muscle fibers with various myosin heavy chain (MHC) isoforms (I, I/IIA, IIA and IIAX) across a range of Ca2+ concentrations in 11 young (24.1±1.1years) and 10 older (68.8±0.8years) men and women. The normalized pCa-force curve shifted rightward with age, leading to decreased activation threshold (pCa10) and/or Ca2+ sensitivity (pCa50) for all MHC isoforms examined. In older adults, the slope of the pCa-force curve was unchanged in MHC I-containing fibers (I, I/IIA), but was steeper in MHC II-containing fibers (IIA, IIAX), indicating greater cooperativity compared to young adults. At sub-maximal [Ca2+], specific force was reduced in MHC I-containing fibers, but was minimally decreased in MHC IIA fibers as older adults produced greater specific forces at high [Ca2+] in these fibers. Lessor pCa50 in MHC I fibers independently predicted reduced isokinetic knee extensor power across a range of contractile velocities, suggesting that the Ca2+ response of slow-twitch fibers contributes to whole muscle dysfunction. Our findings show that aging attenuates Ca2+ responsiveness across fiber types and that these cellular alterations may lead to age-related reductions in whole muscle power output.

Effect of androgen deprivation therapy on the contractile properties of type I and type II skeletal muscle fibres in men with non‐metastatic prostate cancer

The contractile properties of vastus lateralis muscle fibres were examined in prostate cancer (PrCa) patients undergoing androgen deprivation therapy (ADT) and in age- and activity-matched healthy male subjects (Control). Mechanically-skinned muscle fibres were exposed to a sequence of heavily Ca2+ -buffered solutions at progressively higher free [Ca2+ ] to determine their force-Ca2+ relationship. Ca2+ -sensitivity was decreased in both type I and type II muscle fibres of ADT subjects relative to Controls (by -0.05 and -0.04 pCa units, respectively, P<.02), and specific force was around 13% lower in typeI fibres of ADT subjects than in Controls (P=.02), whereas there was no significant difference in type II fibres. Treatment with the reducing agent dithiothreitol slightly increased specific force in type I and type II fibres of ADT subjects (by ~2%-3%, P<.05) but not in Controls. Pure type IIx fibres were found frequently in muscle from ADT subjects but not in Controls, and the overall percentage of myosin heavy chain IIx in muscle samples was 2.5 times higher in ADT subjects (P<.01). The findings suggest that testosterone suppression can negatively impact the contractile properties by (i) reducing Ca2+ -sensitivity in both type I and type II fibres and (ii) reducing maximum specific force in type I fibres.

Effects of pennation angle, electrodes orientation and knee angle on surface electromyography of vastus lateralis during isometric contractions

The effects of pennation angle (PA), electrode orientation (EO) and knee angle (KA) on the amplitude of electromyography signal (EMG) of vastus lateralis muscle (VL) were investigated. Seventeen participants (8 men and 9 women; 26.5 ± 5.1 years; 64.4 ± 12.3 kg; 1.66 ± 0.09 m) accomplished isometric tests at four contraction intensities (COI): 25, 50, 75 and 100% of maximum voluntary contraction (MVC), with KA at 90° and 120°. PA was measured at rest and during contractions. One pair of electrodes was aligned with PA of relaxed VL fibers (pair 1) and another (pair 2) was 20° internally rotated to pair 1. The EMG root mean square (RMS) was calculated and later corrected (RMSC) using equations including PA and 20° angle. PA increased with COI (p 0.05). The PA can be used to improve RMS estimation and further expressed as %MVC maybe promisor in EMG–force related issues. Association between EMG and PA appears to be dependent on their relation to COI.

Ca(2+) leakage out of the sarcoplasmic reticulum is increased in type I skeletal muscle fibres in aged humans.

The amount of Ca(2+) stored in the sarcoplasmic reticulum (SR) of muscle fibres is decreased in aged individuals, and an important question is whether this results from increased Ca(2+) leakage out through the Ca(2+) release channels (ryanodine receptors; RyRs). The present study examined the effects of blocking the RyRs with Mg(2+), or applying a strong reducing treatment, on net Ca(2+) accumulation by the SR in skinned muscle fibres from Old (∼70 years) and Young (∼24 years) adults. Raising cytoplasmic [Mg(2+)] and reducing treatment increased net SR Ca(2+) accumulation in type I fibres of Old subjects relative to that in Young. The densities of RyRs and dihydropyridine receptors were not significantly changed in the muscle of Old subjects. These findings indicate that oxidative modification of the RyRs causes increased Ca(2+) leakage from the SR in muscle fibres in Old subjects, which probably deleteriously affects normal muscle function both directly and indirectly. The present study examined whether the lower Ca(2+) storage levels in the sarcoplasmic reticulum (SR) in vastus lateralis muscle fibres in Old (70 ± 4 years) relative to Young (24 ± 4 years) human subjects is the result of increased leakage of Ca(2+) out of the SR through the Ca(2+) release channels/ryanodine receptors (RyRs) and due to oxidative modification of the RyRs. SR Ca(2+) accumulation in mechanically skinned muscle fibres was examined in the presence of 1, 3 or 10 mm cytoplasmic Mg(2+) because raising [Mg(2+)] strongly inhibits Ca(2+) efflux through the RyRs. In type I fibres of Old subjects, SR Ca(2+) accumulation in the presence of 1 mm Mg(2+) approached saturation at shorter loading times than in Young subjects, consistent with Ca(2+) leakage limiting net uptake, and raising [Mg(2+)] to 10 mm in such fibres increased maximal SR Ca(2+) accumulation. No significant differences were seen in type II fibres. Treatment with dithiothreitol (10 mm for 5 min), a strong reducing agent, also increased maximal SR Ca(2+) accumulation at 1 mm Mg(2+) in type I fibres of Old subjects but not in other fibres. The densities of dihydropyridine receptors and RyRs were not significantly different in muscles of Old relative to Young subjects. These findings indicate that Ca(2+) leakage from the SR is increased in type I fibres in Old subjects by reversible oxidative modification of the RyRs; this increased SR Ca(2+) leak is expected to have both direct and indirect deleterious effects on Ca(2+) movements and muscle function.