Sol Muscles Research Articles

Reliability of In Vivo Creatine-Weighted Chemical Exchange Saturation Transfer (CrCEST) MRI in Calf Skeletal Muscle of Healthy Volunteers at 3 T.

Skeletal muscle mitochondrial oxidative phosphorylation (mtOXPHOS) is important for ATP generation and its dysfunction leads to exercise intolerance. Phosphorus magnetic resonance spectroscopy (31P-MRS) is a useful, noninvasive technique for mtOXPHOS assessment but has limitations. Creatine-weighted chemical exchange saturation transfer (CrCEST) MRI is a potential alternative to assess muscle bioenergetics. To evaluate the interscan repeatability, intra- and interobserver reproducibility of CrCEST during mild plantar flexion exercise. Retrospective. Twenty healthy volunteers (age 37.6 ± 12.4 years, 11 females). 3 T/CEST imaging using gradient echo readout. τCrCEST (postexercise Cr recovery time) was assessed in two scans for each participant, following mild plantar flexion exercises targeting the medial gastrocnemius (MG), lateral gastrocnemius (LG), and soleus (Sol) muscles. Three observers measured τCrCEST for interobserver reproducibility. Three readings by one observer were used to measure intraobserver reproducibility. Two scans were used for within-participant interscan repeatability. Paired t tests, intraclass correlation coefficient (ICC), and Pearson correlation were conducted. Bland-Altman plots were used to analyze the interobserver variability. A P-value of 0.05 was considered statistically significant. There was excellent intra- (ICC ) and interobserver (ICC ) reproducibility, with moderate interscan repeatability for τCrCEST in LG and MG (ICC ) and poor-to-moderate interscan repeatability in Sol (ICC ). Excellent interobserver reproducibility was confirmed by Bland-Altman plots (fixed bias P-value ). CrCEST MRI shows promise in assessing muscle bioenergetics by evaluating τCrCEST during mild plantar flexion exercise with reasonable reliability, particularly in LG and MG. 4 TECHNICAL EFFICACY: Stage 1.

Alterations in sphingolipid synthesis during training, detraining, and retraining may alter myogenesis in skeletal muscle

The benefits of exercise on metabolic health and its effectiveness at reducing morbidity risk are well established. Exercise training enhances skeletal muscle metabolism and function, but whether such adaptations persist through detraining and how they are potentiated with retraining are not well understood. Recent evidence has highlighted the role sphingolipids play on skeletal muscle size and function, but how exercise affects sphingolipid metabolism is not yet known. We hypothesize that alterations in skeletal muscle sphingolipid metabolism with exercise training are necessary to enhance muscle mass with further exercise retraining. Here, mice were SEDentary (SED; static cages) or exercise-TRAINed (TRAIN; voluntary wheel running) and chow-fed. We performed body composition analysis (using EchoMRI) and skeletal muscle collection [tibialis anterior ( TA), extensor digitorum longus ( EDL), gastrocnemius ( Gas), Soleus ( Sol), plantaris ( Plant), and levator ani ( LA)] following training (4wks), detraining (8wk), and retraining (12wk) in three independent cohorts. Sphingolipid and myogenic gene expression was measured in TA muscle by qPCR. Exercise training concomitantly increased lean and decreased fat percentage ( p<0.05), while increasing Sol muscle mass in TRAIN mice ( p<0.05), compared to SED. This was accompanied by higher sphingolipid metabolism ( Acer1, p<0.05; and Cers1, p=0.058) and myogenic gene expression ( MyoD, Myf5; p<0.05). Following detraining, TRAIN mice had an overall increase in lean-to-fat mass ( p<0.05) while maintaining all individual muscle masses ( p<0.05), compared to SED. This was accompanied by a reduction in gene expression of the sphingolipid receptor S1pr1 and myogenic factor Myf5 ( p<0.05), but no differences in sphingolipid metabolism ( Acer1, Cers1). Lastly, retraining increased lean and decreased fat percentage ( p<0.05) and enhanced the masses of TA, Gas, Sol, and Plant muscles in TRAIN mice ( p<0.05), compared to SED. This was accompanied by a reduced expression of rate limiting sphingolipid synthesis genes Sptlc1 and Sptlc2 and the kinase Sphk1 ( p<0.05), but no differences in Cers1 or myogenic gene expression in TRAIN mice, compared to SED. Overall, our data shows that exercise training and retraining potentiate muscle mass accretion and dynamically regulates sphingolipid metabolism adaptations. These data suggest a potential role for sphingolipids to modulate long-lasting improvements to skeletal muscle. Funding was provided by the University of Illinois Urbana-Champaign. This is the full abstract presented at the American Physiology Summit 2024 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.

Investigation on Transient Receptor Potential Vanilloid 1 (TRPv1) Activation in Skeletal Muscle: Pilot Studies

Transient Receptor Potential Vanilloid 1 (TRPv1) is a Ca2+ ion channel predominately studied in neuronal cells and has been found to have sex-differential expression in mouse arterial smooth muscle. More recently, TRPv1 expression has also been observed in skeletal muscle. Among other stimuli, osmotic stress has shown to activate TRPv1 in different tissues (e.g., retinal ganglion and supraoptic nuclear cells). However, the effect of hyperglycemia- and dehydration-induced osmotic stress and the sex-differential expression on TRPv1 in skeletal muscle is not known. PURPOSES: To examine the effect of sex ( Pilot 1), hyperglycemia ( Pilot 2), and dehydration ( Pilot 3) on TRPv1 in skeletal muscle. HYPOTHESES: We hypothesized that TRPv1 would be different between sexes ( Pilot 1), would be higher in hyperglycemic rats compared to healthy controls ( Pilot 2), and would be greater in dehydrated than hydrated state in men ( Pilot 3). METHODS: To accomplish these purposes, we analyzed a subset of samples from three previously completed projects. Pilot 1: Vastus lateralis (VL) muscles were collected from untrained men ( n=3) and women ( n=3) at baseline. Pilot 2: Rats fed a high fat diet and injected with streptozotocin (35 mg/kg) (HFD; n=3; non-fasting blood glucose (NFBG) ≥300mg/dL) were compared to rats fed a normal diet (CON; n=3; NFBG <300mg/dL). Soleus (Sol) muscle was collected after 7 weeks of feeding. Pilot 3: Lastly, VL muscles were collected from trained men (n=5) either dehydrated (DEHY; 296 ± 6 plasma mmol/kg) or hydrated (EUHY; 283 ± 5 plasma mmol/kg) for 24 hours. Rat Sol and human VL samples were analyzed for TRPv1 and p-TRPv1ser502 protein content. TRPv1 activation was calculated as p-TRPv1ser502/ TRPv1/ total protein. Student’s T-tests and effect sizes (ES; small [≤0.49], medium [0.5-0.79], or large [≥0.8] effect) were used to detect differences. Results: Pilot 1: A significant ( p<0.05) difference was observed for TRPv1 with greater expression in men than women (43%, p=0.03, d=2.757). A medium ES was observed for TRPv1 activation (31%, p=0.42, d=0.728) being greater in men compared to women. Pilot 2: No significant difference was observed for TRPv1 and TRPv1 activation between CON and HFD rats. A medium ES was observed for TRPv1 expression (-9%, p=0.31, d=0.441) being lower in HFD compared to CON. A large effect size was observed for TRPv1 activation (30%, p=0.18, d=1.689) being greater in HFD compared to CON. Pilot 3: No significant difference was observed for TRPv1 and TRPv1 activation between EUHY and DEHY. A large ES was observed for TRPv1 (34%, p=0.15, d=0.995) and small ES was observed for TRPv1 activation (2%, p=0.80, d=0.164) being greater in DEHY compared to EUHY. CONCLUSION: These data are the first to illustrate the potential difference in TRPv1 in skeletal muscle. Future research is needed to explore the impact of differential TPRv1 activation on downstream Ca2+ signaling between sexes, dehydration, and hyperglycemic models. This is the full abstract presented at the American Physiology Summit 2024 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.

Sucrose-Enriched and Carbohydrate-Free High-Fat Diets Distinctly Affect Substrate Metabolism in Oxidative and Glycolytic Muscles of Rats.

Skeletal muscle substrate preference for fuel is largely influenced by dietary macronutrient availability. The abundance of dietary carbohydrates promotes the utilization of glucose as a substrate for energy production, whereas an abundant dietary fat supply elevates rates of fatty acid (FA) oxidation. The objective of this study was to determine whether an obesogenic, high-fat, sucrose-enriched (HFS) diet or a carbohydrate-free ketogenic diet (KD) exert distinct effects on fat, glucose, and ketone metabolism in oxidative and glycolytic skeletal muscles. Male Wistar rats were fed either a HFS diet or a KD for 16 weeks. Subsequently, the soleus (Sol), extensor digitorum longus (EDL), and epitrochlearis (Epit) muscles were extracted to measure palmitate oxidation, insulin-stimulated glucose metabolism, and markers of mitochondrial biogenesis, ketolytic capacity, and cataplerotic and anaplerotic machinery. Sol, EDL, and Epit muscles from KD-fed rats preserved their ability to elevate glycogen synthesis and lactate production in response to insulin, whereas all muscles from rats fed with the HFS diet displayed blunted responses to insulin. The maintenance of metabolic flexibility with the KD was accompanied by muscle-fiber-type-specific adaptive responses. This was characterized by the Sol muscle in KD-fed rats enhancing mitochondrial biogenesis and ketolytic capacity without elevating its rates of FA oxidation in comparison with that in HFS feeding. Conversely, in the Epit muscle, rates of FA oxidation were increased, whereas the ketolytic capacity was markedly reduced by the KD in comparison with that by HFS feeding. In the EDL muscle, the KD also increased rates of FA oxidation, although it did so without altering its ketolytic capacity when compared to HFS feeding. In conclusion, even though obesogenic and ketogenic diets have elevated contents of fat and alter whole-body substrate partitioning, these two dietary interventions are associated with opposite outcomes with respect to skeletal muscle metabolic flexibility.

Contractile Work of the Soleus and Biarticular Mechanisms of the Gastrocnemii Muscles Increase the Net Ankle Mechanical Work at High Walking Speeds.

Increasing walking speed is accompanied by an increase of the mechanical power and work performed at the ankle joint despite the decrease of the intrinsic muscle force potential of the soleus (Sol) and gastrocnemius medialis (GM) muscles. In the present study, we measured Achilles tendon (AT) elongation and, based on an experimentally determined AT force-elongation relationship, quantified AT force at four walking speeds (slow 0.7 m.s-1, preferred 1.4 m.s-1, transition 2.0 m.s-1, and maximum 2.6 ± 0.3 m.s-1). Further, we investigated the mechanical power and work of the AT force at the ankle joint and, separately, the mechanical power and work of the monoarticular Sol at the ankle joint and the biarticular gastrocnemii at the ankle and knee joints. We found a 21% decrease in maximum AT force at the two higher speeds compared to the preferred; however, the net work of the AT force at the ankle joint (ATF work) increased as a function of walking speed. An earlier plantar flexion accompanied by an increased electromyographic activity of the Sol and GM muscles and a knee-to-ankle joint energy transfer via the biarticular gastrocnemii increased the net ATF mechanical work by 1.7 and 2.4-fold in the transition and maximum walking speed, respectively. Our findings provide first-time evidence for a different mechanistic participation of the monoarticular Sol muscle (i.e., increased contractile net work carried out) and the biarticular gastrocnemii (i.e., increased contribution of biarticular mechanisms) to the speed-related increase of net ATF work.

Insulin-resistant female rat skeletal muscles display diacylglycerol-mediated protein kinase C activation and inflammation without ceramide accumulation.

This study investigated the role of diacylglycerol (DAG)-mediated protein kinase C (PKC) activation, ceramide accumulation and inflammation in insulin-resistant female oxidative and glycolytic skeletal muscles induced by an obesogenic high-fat sucrose-enriched (HFS) diet. The HFS diet impaired insulin-stimulated AKTThr308 phosphorylation and glycogen synthesis, whereas rates of fatty acid oxidation and basal lactate production were significantly elevated in soleus (Sol), extensor digitorum longus (EDL) and epitrochlearis (Epit) muscles. Insulin resistance was accompanied by increases in triacylglycerol (TAG) and DAG contents in Sol and EDL, whereas in Epit muscles only TAG content and markers of inflammation were associated with HFS diet-induced insulin resistance. Analysis of membrane-bound/cytoplasmic PKC fractions revealed that the HFS diet promoted activation/translocation of PKCδ and θ isoforms in Sol, EDL and Epit muscles. However, none of these muscles displayed alterations in ceramide content in response to HFS feeding. This could be explained by a significant increase in Dgat2 mRNA expression in Sol, EDL and Epit muscles, which likely diverted most of the intramyocellular acyl-CoAs toward TAG synthesis instead of ceramides. Overall, this study helps elucidate the molecular mechanisms underlying insulin resistance caused by diet-induced obesity in female skeletal muscles with distinct fibre type compositions. KEY POINTS: Feeding female Wistar rats a high-fat sucrose-enriched diet (HFS) led to diacylglycerol (DAG)-induced PKC activation and insulin resistance in oxidative and glycolytic skeletal muscles. HFS diet-induced toll-like receptor 4 (Tlr4) expression did not lead to increased ceramide content in female skeletal muscles. In highly glycolytic female muscles, elevated TAG content and markers of inflammation underlay HFS diet-induced insulin resistance. The HFS diet suppressed glucose oxidation and increased lactate production in oxidative and glycolytic female muscles. Increased Dgat2 mRNA expression likely diverted most of the intramyocellular acyl-CoAs toward TAG synthesis and prevented ceramide formation in skeletal muscles of HFS-fed female rats.

Phosphoproteomic analysis identifies differentially expressed phosphorylation sites that affect muscle fiber type in pigs.

Skeletal muscle of livestock is composed of both fast- and slow-twitch muscle fibers, which are key factors in their meat quality. However, the role of protein phosphorylation in muscle fiber type is not completely understood. Here, a fast-twitch (biceps femoris, BF) and slow-twitch (soleus, SOL) muscle tissue sample was collected from three male offspring of Duroc and Meishan pigs. We demonstrate that the meat quality of SOL muscle is significantly better than that of BF muscle. We further used phosphoproteomic profiling of BF and SOL muscles to identify differences between these muscle types. A total of 2,327 phosphorylation sites from 770 phosphoproteins were identified. Among these sites, 287 differentially expressed phosphorylation sites (DEPSs) were identified between BF and SOL. GO and KEGG enrichment analysis of proteins containing DEPSs showed that these phosphorylated proteins were enriched in the glycolytic process GO term and the AMPK signaling pathway. A protein-protein interaction (PPI) analysis reveals that these phosphorylated proteins interact with each other to regulate the transformation of muscle fiber type. These analyses reveal that protein phosphorylation modifications are involved in porcine skeletal muscle fiber type transformation. This study provides new insights into the molecular mechanisms by which protein phosphorylation regulates muscle fiber type transformation and meat quality in pigs.

BDNF Spinal Overexpression after Spinal Cord Injury Partially Protects Soleus Neuromuscular Junction from Disintegration, Increasing VAChT and AChE Transcripts in Soleus but Not Tibialis Anterior Motoneurons.

After spinal cord transection (SCT) the interaction between motoneurons (MNs) and muscle is impaired, due to reorganization of the spinal network after a loss of supraspinal inputs. Rats subjected to SCT, treated with intraspinal injection of a AAV-BDNF (brain-derived neurotrophic factor) construct, partially regained the ability to walk. The central effects of this treatment have been identified, but its impact at the neuromuscular junction (NMJ) has not been characterized. Here, we compared the ability of NMJ pre- and postsynaptic machinery in the ankle extensor (Sol) and flexor (TA) muscles to respond to intraspinal AAV-BDNF after SCT. The gene expression of cholinergic molecules (VAChT, ChAT, AChE, nAChR, mAChR) was investigated in tracer-identified, microdissected MN perikarya, and in muscle fibers with the use of qPCR. In the NMJs, a distribution of VAChT, nAChR and Schwann cells was studied by immunofluorescence, and of synaptic vesicles and membrane active zones by electron microscopy. We showed partial protection of the Sol NMJs from disintegration, and upregulation of the VAChT and AChE transcripts in the Sol, but not the TA MNs after spinal enrichment with BDNF. We propose that the observed discrepancy in response to BDNF treatment is an effect of difference in the TrkB expression setting BDNF responsiveness, and of BDNF demands in Sol and TA muscles.

Muscle Atrophy Evaluation via Radiomics Analysis Using Ultrasound Images: A Cohort Data Study.

Existing methods for muscle atrophy evaluation based on muscle size measures from ultrasound images are inadequate in precision. Radiomics has been widely used in various medical studies, but its validity for the evaluation of muscle atrophy has not been fully explored. This study presents a radiomics analysis for muscle atrophy evaluation using ultrasound images. The hindlimb unloading rat model was developed to simulate weightlessness muscle atrophy and ultrasound images of the hind limbs were acquired for both the hindlimb unloaded (HU) and control groups during a 21-day HU period. A total of 368 radiomics features were extracted and the stable and informative features were selected through a two-stage feature selection procedure. The feature change trajectory of the stable features was analyzed using the hierarchical clustering method. Finally, an adaptive longitudinal feature selection and grading network, ALNet, was developed to evaluate muscle atrophy. The clustering trajectories of ultrasound image features showed similar trends to the changes in muscle atrophy at the molecular level. The best grading accuracy achieved by the ALNet was 79.5% for the Soleus (Sol) muscle and 82.6% for the Gastrocnemius (Gas) muscle. The test-retest is essential in performing radiomics analysis on ultrasound images. The longitudinal feature selection is important for muscle atrophy grading. The ultrasound image features of the Gas muscle have better discrimination ability than that of the Sol muscle. This study proves for the first time the capability of ultrasound image features for muscle atrophy evaluation.

Skeletal muscle contraction kinetics and AMPK responses are modulated by the adenine nucleotide degrading enzyme AMPD1.

AMP deaminase 1 (AMPD1; AMP → IMP + NH3) deficiency in skeletal muscle results in an inordinate accumulation of AMP during strenuous exercise, with some but not all studies reporting premature fatigue and reduced work capacity. To further explore these inconsistencies, we investigated the extent to which AMPD1 deficiency impacts skeletal muscle contractile function of different muscles and the [AMP]/AMPK responses to different intensities of fatiguing contractions. To reduce AMPD1 protein, we electroporated either an inhibitory AMPD1-specific miRNA encoding plasmid or a control plasmid, into contralateral EDL and SOL muscles of C57BL/6J mice (n = 48 males, 24 females). After 10 days, isolated muscles were assessed for isometric twitch, tetanic, and repeated fatiguing contraction characteristics using one of four (None, LOW, MOD, and HIGH) duty cycles. AMPD1 knockdown (∼35%) had no effect on twitch force or twitch contraction/relaxation kinetics. However, during maximal tetanic contractions, AMPD1 knockdown impaired both time-to-peak tension (TPT) and half-relaxation time (½ RT) in EDL, but not SOL muscle. In addition, AMPD1 knockdown in EDL exaggerated the AMP response to contractions at LOW (+100%) and MOD (+54%) duty cycles, but not at HIGH duty cycle. This accumulation of AMP was accompanied by increased AMPK phosphorylation (Thr-172; LOW +25%, MOD +34%) and downstream substrate phosphorylation (LOW +15%, MOD +17%). These responses to AMPD1 knockdown were not different between males and females. Our findings demonstrate that AMPD1 plays a role in maintaining skeletal muscle contractile function and regulating the energetic responses associated with repeated contractions in a muscle- but not sex-specific manner.NEW & NOTEWORTHY AMP deaminase 1 (AMPD1) deficiency has been associated with premature muscle fatigue and reduced work capacity, but this finding has been inconsistent. Herein, we report that although AMPD1 knockdown in mouse skeletal muscle does not change maximal isometric force, it negatively impacts muscle function by slowing contraction and relaxation kinetics in EDL muscle but not SOL muscle. Furthermore, AMPD1 knockdown differentially affects the [AMP]/AMPK responses to fatiguing contractions in an intensity-dependent manner in EDL muscle.

Triceps surae muscle force potential and force demand shift with altering stride frequency in running.

While it is well recognized that the preferred stride frequency (PSF) in running closely corresponds to the metabolically optimal frequency, the underlying mechanisms are still unclear. Changes in joint kinematics when altering stride frequency will affect the muscle-tendon unit lengths and potentially the efficiency of muscles crossing these joints. Here, we investigated how fascicle kinematics and forces of the triceps surae muscle, a highly energy consuming muscle, are affected when running at different stride frequencies. Twelve runners ran on a force measuring treadmill, adopting five different frequencies (PSF; PSF ± 8%; PSF ± 15%), while we measured joint kinematics, whole-body energy expenditure, triceps surae muscle activity, and soleus (SOL; N=10) and gastrocnemius medialis (GM; N=12) fascicle kinematics. In addition, we used dynamic optimization to estimate SOL and GM muscle forces. We found that SOL and GM mean muscle fascicle length during stance followed an inverted U-relationship with the longest fascicle lengths occurring at PSF. Fascicle lengths were shortest at frequencies lower than PSF. In addition, average SOL force was greater at PSF-15% compared with PSF. Overall, our results suggest that reduced SOL and GM muscle fascicle lengths, associated with reduced muscle force potential, together with greater SOL force demand, contribute to the increased whole-body energy expenditure when running at lower than PSF. At higher stride frequencies, triceps surae muscle kinematics and force production were less affected suggesting that increased energy expenditure is rather related to higher cost of leg swing and greater cost of force production.

Expression profiles of protein markers regulating the mitochondrial lifecycle in skeletal muscle of acute spinal cord transected rats

Spinal cord transection (ST) inactivates the neuromuscular complex and triggers progressive muscular atrophy of the affected skeletal muscles. Disruption in neural input also leads to a reduction in mitochondrial volume. The mitochondria lifecycle can be generally divided into processes of biogenesis, expression of OXPHOS proteins, fusion, fission, and mito/autophagy; specifically, the balance between biogenesis vs. mitophagy dictates the rate of formation and destruction, respectively. Here, we examined the expression profiles of the markers associated with the mitochondrial lifecycle up to one month following ST in phenotypically slow (soleus; SOL) and mixed (plantaris; PLT) skeletal muscle. We hypothesized that ST would induce a reduction in mitochondrial DNA (mtDNA) copy number, decrease expression in markers associated with biogenesis, and increased proteins that regulate mito/autophagy. Adult female Sprague Dawley rats were randomly divided into control (CON; n=6), 1 day (1dST; n=5), 8 day (8dST; n=8), and 28 day post‐ST (28dST; n=8). Compared to CON relative SOL and PLT muscle masses (absolute mass / body mass) were 98, 65, and 62% and 85, 60, and 73% at 1, 8 and 28d post‐ST, respectively (p<0.05). Compared to CON, no significant changes were observed in mtDNA copy number in ST muscles following amplification using standard end‐point PCR protocols for mtDNA (ND1, COX1, or ATP6) and nuclear (β2M) genes (p>0.05). Total protein was isolated from SOL and PLT, separated using standard SDS‐PAGE protocol, and probed for markers associated with various stages of the mitochondrial lifecycle using standard western blot protocols. Upstream markers known to influence mitochondrial lifecycle signaling (Rev‐Erbα, AMPK) showed a varied temporal and muscle‐specific response: in 1dST SOL muscles, Rev‐Erbα increased 74% from CON then decreased 43% at 28dST, whereas 8dST PLT muscle exhibited a 44% increase (p<0.05). In 8dST SOL and PLT muscles, the pAMPK:AMPK ratio was elevated 366 and 85% from CON, respectively (p<0.05). Markers for mitochondrial biogenesis (PGC‐1α, NRF1, NRF2) in SOL and PLT muscles were largely unchanged, although a ~50% decrease in Tfam protein expression was observed at each ST time point compared to CON (p<0.05). Mitofusion2, a marker for mitochondrial fusion, was unchanged (p>0.05). However, for markers detecting mitochondrial fission (MFF, Drp1, Fis1), we observed a ~60% increase in MFF expression in 1dST SOL and PLT muscles (p<0.05). As expected markers for mitophagy (Parkin, PINK, p62, LC3 I/II) were significantly elevated: Parkin increased ~70% at 28dST (p<0.05), whereas LC3 I/II increased 65% and 112% at 1dST in in SOL and PLT muscles, respectively (p<0.05), and remained significantly elevated in PLT muscles at 8dST (27%) and 28d (37%) (p<0.05). Taken together, these data suggest that mitochondrial volume and expression of markers regulating mitochondrial biogenesis and fusion are not influenced by acute ST despite a decrease in muscle mass. However, selective markers for fission and mitophagy are significantly elevated in both SOL and PLT muscles, suggesting that mitochondrial destruction mechanism are activated acutely following ST injury. Lastly, selective proteins of the OXPHOS protein complex significantly increased in 1dST SOL and PLT muscles suggesting possible activity‐dependent regulation of mitochondrial gene expression.

Contribution of sensory feedback to soleus muscle activity during voluntary contraction in humans

Sensory feedback through spinal interneurons contributes to plantar flexor muscle activity during walking, but it is unknown whether this is also the case during nonlocomotor movements. Here, we explored the effect of temporary reduction of sensory feedback to ankle plantar flexors during voluntary contraction in sitting subjects. Thirteen healthy adults (mean age 32 yr) were seated with the right leg attached to a foot plate which could be moved in dorsi- or plantarflexion direction by a computer-controlled motor. EMG was recorded from the tibialis anterior (TA) and soleus (Sol) muscles. During static plantar flexion, while the plantar flexors were slowly stretched, a sudden plantar flexion caused a decline in Sol EMG at the same latency as the stretch reflex. This decline in EMG activity was still observed when transmission from dorsiflexors was blocked. It disappeared when transmission from ankle plantar flexors was also blocked. The same quick plantarflexion failed to produce a decline in EMG activity at the latency of the stretch reflex in the absence of slow stretch of the plantar flexors. Instead, a decline in EMG activity was observed 15-20 ms later. This decline disappeared following block of transmission from antagonists, suggesting that reciprocal inhibition was involved. These findings show that unload of ankle plantar flexors does not cause a similar drop in Sol EMG during voluntary contraction as during walking. This implies that sensory feedback through spinal interneurons only contributes little to the neural drive to plantar flexor muscles during human voluntary contraction in sitting subjects.NEW & NOTEWORTHY Sensory feedback through spinal reflex pathways makes only a minor contribution to neural drive to muscles during voluntary ankle plantar flexion. This differs distinctly from observations during walking and suggests that the neural drive to ankle plantar flexors during voluntary contraction do not rely on sensory feedback through similar spinal interneuronal networks as during walking. In line with animal studies this suggests that the integration of sensory feedback in CNS is task specific.

How fiber dynamics of plantarflexor and dorsiflexor muscles based on EMG-driven approach can explain the metabolic cost at different gait speeds.

The aim of this study was to investigate the fiber dynamics of plantarflexor and dorsiflexor muscles and their association with the net metabolic rate (NCw). Metabolic, kinematic, kinetic, and electromyography measurements were made on seven young subjects while they walked on a force-plate instrumented treadmill at 1.00, 1.20, 1.40, 1.60, and 1.8m/s for 1:30 min. The net metabolic rate was computed, and a one degree-of freedom EMG-driven approach was used to extract the force generation ability (Fability), and active force-length (fAL) and force-velocity (fV) multiplier of each muscle. A one-way (speeds) repeated measures ANOVA was performed for each muscle and a multiple linear regression model was used to explain NCw. Fability was significantly affected by gait speed for the GasMed and the SOL muscles. The decrease of Fability for the SOL and the GasMed was accompanied by a decrease in the force-velocity multiplier. The peak muscle force for the SOL increased for the lowest speed compared to the higher speed, and for the TibAnt increased at high speed compared to low speed. In addition, Fability fAL, and fV of the SOL predicted over 58% of NCw and FMax of the TibAnt accounts for 39.9% of the variance in NCw. The increase of NCw with gait speed over the preferred walking speed can be partially explained by the decreasing capacity of the SOL muscle to produce muscle force and more specifically by the force-velocity relationship and an increase in muscle force for the TibAnt.

Automated segmentation of the gastrocnemius and soleus in shank ultrasound images through deep residual neural network

Segmentation of the shank gastrocnemius (Gas) and soleus (Sol) muscles in ultrasound (US) images allows to extract the muscle features, which are important for the early diagnosis of muscle atrophy. The automatic segmentation of the muscles is a challenging task, and deep learning (DL) provides a solution to this problem, which can effectively extract representative features from the muscle regions and background of the images. In this study, we propose ResTU-net, an automatic segmentation method based on improved U-net network, to segment the Gas and Sol muscles in shank US images. This network uses the deep residual neural network (Resnet) as the sublayer unit of each layer of a U-net, and can effectively combine the features of each layer with those of the next layer to meet the challenges of poor US image quality and low contrast. In addition, dilated convolution is used instead of the pooling layer in the network to prevent information loss during training. Experiments were performed on 3350 shank US images from 23 Sprague Dawley (SD) rats, among them, 2650 shank US images were used for network training and 700 for network validation. Compared with state-of-the-art networks, the experimental results show that the method can achieved the best segmentation capability results and a mean Dice similarity coefficient (DSC) of the Gas and Sol muscles of 94.82% and 90.72%, respectively. This work indicates that the proposed fully automatic segmentation method may be accurately and efficiently applied to Gas and Sol muscles segmentation in shank US images.

Evaluation of muscle-specific and metabolism regulating microRNAs in a chronic swimming rat model

Making benefit from the epigenetic effects of environmental factors such as physical activity may result in a considerable improvement in the prevention of chronic civilization diseases. In our chronic swimming rat model, the expression levels of such microRNAs were characterized, that are involved in skeletal muscle differentiation, hypertrophy and fine-tuning of metabolism, which processes are influenced by chronic endurance training, contributing to the metabolic adaptation of skeletal muscle during physical activity. After chronic swimming, the level of miR-128a increased significantly in EDL muscles, which may influence metabolic adaptation and stress response as well. In SOL, the expression level of miR-15b and miR-451 decreased significantly after chronic swimming, which changes are opposite to their previously described increment in insulin resistant skeletal muscle. MiR-451 also targets PGC-1α mRNA, whiches expression level significantly increased in SOL muscles, resulting in enhanced biogenesis and oxidative capacity of mitochondria. In summary, the microRNA expression changes that were observed during our experiments suggest that chronic swim training contributes to a beneficial metabolic profile of skeletal muscle.

Variation in muscle and neuromuscular junction morphology between atrophy-resistant and atrophy-prone muscles supports failed re-innervation in aging muscle atrophy.

In advanced age, there is an accelerated decline in skeletal muscle mass that appears to be secondary to repeated cycles of denervation-reinnervation and eventually, failed reinnervation. However, whether variation in reinnervation capacity explains why some muscles are less vulnerable to age-related atrophy has not been addressed. In this study we examined changes in neuromuscular junction (NMJ) morphology, fiber cross-sectional area (CSA) and fiber type, accumulation of severely atrophied myofibers, and expression of a marker of denervation in four muscles that exhibit differences in the degree of age-related atrophy and which span the extremes of fiber type composition in 8 mo old (8M) and 34 mo old (34M) male Fischer 344 Brown Norway F1 hybrid rats. Aging muscle atrophy was most pronounced in the fast twitch gastrocnemius (Gas; 25%) and similar between extensor digitorum longus (EDL) and slow-twitch soleus (Sol) muscle (14-15%), whereas the slow-twitch adductor longus (AL) increased in mass by 21% between 8M and 34M (P<0.05 for all). Only the Sol exhibited significant alterations in fiber type with aging, and there was a decrease in fiber CSA in the Gas, EDL, and Sol (P<0.05) with aging that was not seen in the AL. Muscles that atrophied had an increased fraction of severely atrophic myofibers (P<0.05), but this was not observed in the AL. The Gas and EDL both demonstrated a similar degree of age-related remodeling of pre- and post-synaptic NMJ components. On the other hand, pre- and post-synaptic morphology underwent greater changes with aging in the AL, and many of these same morphological variables were already greater in the Sol vs AL at 8M, suggesting the Sol had already undergone substantial remodeling and may be nearing its adaptive limits. Consistent with this idea, analysis of NMJ morphology in Sol from 3M rats exhibited similar values as 8M AL, and the Sol demonstrated greater expression of the denervation marker neural cell adhesion molecule (NCAM) compared to the AL at 34M. Collectively, our results are consistent with NMJ remodeling capacity being finite with aging and that maintained remodeling potential confers atrophy protection in aging skeletal muscle by reducing the degree of persistent denervation.

Distinct mechanisms involving diacylglycerol, ceramides, and inflammation underlie insulin resistance in oxidative and glycolytic muscles from high fat-fed rats

This study investigated whether oxidative and glycolytic rat skeletal muscles respond differently to a high-fat (HF) sucrose-enriched diet with respect to diacylglycerol (DAG) and ceramides accumulation, protein kinase C (PKC) activation, glucose metabolism, and the expression of inflammatory genes. HF diet (8 weeks) suppressed insulin-stimulated glycogen synthesis and glucose oxidation in soleus (Sol), extensor digitorum longus (EDL) and epitrochlearis (Epit) muscles. However, DAG and ceramides levels increased in Sol and EDL, but not in Epit muscles of HF-fed rats. Additionally, membrane-bound PKC-delta and PKC-theta increased in Sol and EDL, whereas in Epit muscles both PKC isoforms were reduced by HF diet. In Epit muscles, HF diet also increased the expression of tumor necrosis factor-α (TNF-α) receptors (CD40 and FAS), toll-like receptor 4 (TLR4), and nuclear factor kappa light polypeptide gene enhancer in B cells (NF-kB), whereas in Sol and EDL muscles the expression of these inflammatory genes remained unchanged upon HF feeding. In conclusion, HF diet caused DAG and ceramides accumulation, PKC activation, and the induction of inflammatory pathways in a fiber type-specific manner. These findings help explain why oxidative and glycolytic muscles similarly develop insulin resistance, despite major differences in their metabolic characteristics and responsiveness to dietary lipid abundance.

Effects of microgravity exposure and fructo-oligosaccharide ingestion on the proteome of soleus and extensor digitorum longus muscles in developing mice

Short-chain fatty acids produced by the gut bacterial fermentation of non-digestible carbohydrates, e.g., fructo-oligosaccharide (FOS), contribute to the maintenance of skeletal muscle mass and oxidative metabolic capacity. We evaluated the effect of FOS ingestion on protein expression of soleus (Sol) and extensor digitorum longus muscles in mice exposed to microgravity (μ-g). Twelve 9-week-old male C57BL/6J mice were raised individually on the International Space Station under μ-g or artificial 1-g and fed a diet with or without FOS (n = 3/group). Regardless of FOS ingestion, the absolute wet weights of both muscles tended to decrease, and the fiber phenotype in Sol muscles shifted toward fast-twitch type following μ-g exposure. However, FOS ingestion tended to mitigate the μ-g-exposure-related decrease in oxidative metabolism and enhance glutathione redox detoxification in Sol muscles. These results indicate that FOS ingestion mildly suppresses metabolic changes and oxidative stress in antigravity Sol muscles during spaceflight.

BCAAs and Di-Alanine supplementation in the prevention of skeletal muscle atrophy: preclinical evaluation in a murine model of hind limb unloading

Skeletal muscle atrophy occurs in response to various pathophysiological stimuli, including disuse, aging, and neuromuscular disorders, mainly due to an imbalance of anabolic/catabolic signaling. Branched Chain Amino Acids (BCAAs: leucine, isoleucine, valine) supplements can be beneficial for counteracting muscle atrophy, in virtue of their reported anabolic properties. Here, we carried out a proof-of-concept study to assess the in vivo/ex vivo effects of a 4-week treatment with BCAAs on disuse-induced atrophy, in a murine model of hind limb unloading (HU). BCAAs were formulated in drinking water, alone, or plus two equivalents of L-Alanine (2 ALA) or the dipeptide L-Alanyl-L-Alanine (Di-ALA), to boost BCAAs bioavailability. HU mice were characterized by reduction of body mass, decrease of soleus – SOL – muscle mass and total protein, alteration of postural muscles architecture and fiber size, dysregulation of atrophy-related genes (Atrogin-1, MuRF-1, mTOR, Mstn). In parallel, we provided new robust readouts in the HU murine model, such as impaired in vivo isometric torque and ex vivo SOL muscle contractility and elasticity, as well as altered immune response. An acute pharmacokinetic study confirmed that L-ALA, also as dipeptide, enhanced plasma exposure of BCAAs. Globally, the most sensitive parameters to BCAAs action were muscle atrophy and myofiber cross-sectional area, muscle force and compliance to stress, protein synthesis via mTOR and innate immunity, with the new BCAAs + Di-ALA formulation being the most effective treatment. Our results support the working hypothesis and highlight the importance of developing innovative formulations to optimize BCAAs biodistribution.