Tibialis Anterior Research Articles

Muscle Activation Frequency Relationship with Cost of Transport at Increasing Walking Speed in Preliminary Study Reveals Interplay of Both Active and Passive Dynamics

Metabolic cost plays a critical role in gait biomechanics, particularly in rehabilitation. Several factors influence metabolic cost during walking. Therefore, this study aimed to examine the relationship between metabolic cost and muscle activity, focusing on the frequency of EMG signals during walking. We recruited nine participants (five male and four female, age range 20–48 years) who walked for four minutes at six different speeds (ranging from 1.8 to 5.9 mph). EMG data were collected from the dominant lower leg muscles, specifically the lateral gastrocnemius (GAS-L) and the anterior tibialis (AT). Oxygen respiration was measured using open-circuit spirometry. Energy expenditure was estimated as the cost of transport (COT). The EMG data were analyzed using frequency domain features, such as the area under the curve of power spectral density (PSD-AUC) and the maximal distance between two points before and after the peak of the power spectral density curve (MDPSD). The results indicated that PSD-AUC is a better measure than MDPSD for understanding the relationship between activation frequency and COT. PSD-AUC demonstrated an increasing curvilinear trend (R2 = 0.93 and 0.77, second order polynomial fit), but the AT displayed higher variability. MDPSD exhibited more nonlinearity (R2 = 0.17–28, second order polynomial fit), but MDPSD demonstrated statistically significant differences (p < 0.05, t-test independent) in frequency between the GAS-L (64–237 Hz) and AT (114–287 Hz) during slow walking. Additionally, the relationship between COT and PSD-AUC revealed a U-shaped curve, suggesting that high COT is a function of both active and passive dynamics during walking. These findings will be valuable in rehabilitating individuals who suffer from gait-related disorders, especially those related to muscle inefficiency.

Lifespan Effects of Muscle-specific Androgen Receptor Overexpression on Body Composition of Male and Female Rats

Androgenic actions of gonadal testosterone are thought to be a major mechanism promoting sex differences in body composition across the lifespan. However, this inference is based largely on studies of androgen receptor (AR) function in late adolescent or emerging adult rodents. Here we assess body composition and quantify AR expression in skeletal muscle of rats at defined ages, comparing wild-type (WT) to transgenic HSAAR rats which overexpress AR in skeletal muscle. Male and female HSAAR and WT Sprague Dawley rats (N=288) underwent DEXA scanning and tissue collection at post-natal day (PND) 1, 10, 21, 42, 70, 183, 243, and 365. As expected, sex differences in body composition and muscle mass largely onset with puberty (PND-21), with no associated changes to skeletal muscle AR protein. In adulthood, HSAAR increased tibialis anterior (TA) and EDL mass in males, and reduced the expected gain in gonadal fat mass in both sexes. In WT rats, AR protein was reduced in soleus, but not TA, throughout life. Nonetheless, soleus AR protein expression was greater in males than females at all ages of sexual development, yet only at PND-70 in TA. Overall, despite muscle AR overexpression effects, results are inconsistent with major sex differences in body composition during sexual development being driven by changes in muscle AR, but rather suggest that changes in ligand promote sexual differentiation of body composition during pubertal timing. Nonetheless, increased skeletal muscle AR in adulthood can be suffcient to increase muscle mass in males, and reduce adipose in both sexes. This work was supported by an NSERC Grant awarded to D.A.M. (RGPIN 312458). 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.

Heat therapy preconditioning improves the skeletal muscle contractile and mitochondrial function in a mouse model of access-related hand dysfunction

Introduction: Hemodialysis via arteriovenous fistula (AVF) is a life-line treatment for patients with end-stage kidney disease. However, hemodialysis access-related hand dysfunction (ARHD) including ischemia, muscle weakness and alterations in mitochondrial function is prevalent following AVF placement. We tested the hypothesis that two-weeks of daily heat therapy prior to AVF surgery would improve hindlimb function in a mouse model of ARHD. Methods: Adult male C57BL/6J mice (N=9-11/group) were fed a 0.2% adenine diet to induce chronic kidney disease. Following two weeks of the initial adenine diet, animals were randomly allocated to heat therapy (HT) or control (CON) groups. Animals assigned to HT were exposed to environmental heat at 40°C in a climate chamber for 30 min/day, six days/week, for two weeks while the control mice were exposed to room temperature (~22°C). The AVF was created using the left common iliac artery and vein 24-48 hours after the last treatment session. Laser Doppler perfusion to the ventral paw was assessed post-operatively (POD) and AVF patency was verified using a high-frequency duplex ultrasound system three days prior to euthanasia. Following 14 days of recovery, in situ tibialis anterior (TA) muscle contractility and gastrocnemius muscle mitochondrial function were assessed. RNA sequencing analysis of the hindlimb skeletal muscle (N=4/group) was performed. Results: Compared to baseline, the level of blood urea nitrogen was similarly elevated ( P<0.05) in both groups following four weeks of adenine diet. In both groups, blood perfusion to the paw was decreased by ~50% immediately after AVF creation and progressively recovered to ~80% at POD13 without a treatment effect ( P=0.4467). No differences in body weight and muscle mass in the TA, extensor digitorum longus, and soleus muscles were observed (all P>0.05). Notably, however, the absolute ( P=0.0250) and specific force ( P=0.0101) frequency relationship of the TA muscle were significantly improved in the animals exposed to HT. In addition, compared with control animals, HT mice had significant improvement in oxidative phosphorylation (OXPHOS) in both pyruvate ( P=0.0145) and octanoylcarnitine ( P=0.0196) fueled conditions. RNA sequencing results revealed that genes involved in nervous system development and metabolism were significantly different between HT and CON groups. Conclusion: Daily application of heat therapy for two weeks prior to the AVF surgery has beneficial effects on both skeletal muscle contractile and mitochondrial function suggesting this pre-habilitation strategy may mitigate the risk of ARHD in patients undergoing AVF surgery. Funding Support: National Institutes of Health grant R01HL148597 and American Heart Association grant POST903198. 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.

Comparing changes to the skeletal muscle transcriptome of young and aged mice to young and aged humans in response to acute eccentric contractions

Resistance exercise alters several molecular events in skeletal muscle that change muscle size and function, including changes to the transcriptome. The signals altering the muscle transcriptome in response to resistance exercise remain ill defined. Defining those regulatory signals in muscle requires non-human models where manipulations can assess cause and effect. The electrically induced, unilateral eccentric contraction model of resistance exercise in rodents is widely used to mimic resistance exercise. However, the transcriptional changes that occur in response to eccentric contractions in the rodent model and the continuity of the rodent model to resistance exercise in humans are not well known. The purpose of this study was to define the transcriptional changes in skeletal muscle that occur in response to acute eccentric contractions in young and aged mice and define their continuity to transcriptional changes in the muscle of young and aged humans. Four-month-old and 24-month-old male C57BL/6 mice were subjected to a bout of electrically induced, unilateral eccentric contractions of the tibialis anterior (TA) muscle. RNA was isolated from the TA and subjected to RNA Sequencing. Lists of differentially expressed genes (DEGs) from the TA of young and aged mice were compared to RNA Sequencing datasets generated from the vastus lateralis muscle of young and aged humans following acute eccentric contractions. L andscape I n S ilico deletion A nalysis (Lisa) was used to predict the magnitude of transcription factor regulation of the DEGs. DEGs for mice or humans were categorized into age specific DEGs (specific to young or aged) or DEGs shared by both ages. The number of DEGs for mice or humans were distributed equally to each category, showing an age specific distribution that was conserved in both mice and humans. However, the DEGs in young and aged mice had very little overlap to the DEGs in young and aged humans. Only 113 DEGs were common across all ages and species. Interestingly, the relationship of transcription factors predicted to regulate DEGs between young and aged mice or young and aged humans was very strong. However, the relationship of transcription factors predicted to regulate DEGs of young mice and young humans or aged mice and aged humans was rather weak. In all, these data highlight the consistencies and differences in the muscle transcriptomic response to eccentric contractions. This work was supported in part by NIH R03AG073445 (BSG). 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.

Epigallocatechin gallate protects against fat and muscle atrophy in B16BL6 melanoma-bearing mice on a high-fat diet

AimsEpidemiological evidence indicates that there is a substantial association between body mass index (BMI) and at least ten forms of cancer, including melanoma, and BMI imbalance contributes to the poor survival rate of cancer patients before and after therapy. Nevertheless, few pharmacological studies on models of obesity and cancer have been reported. In this study, we administered epigallocatechin gallate (EGCG) to B16BL6 tumor-bearing mice that received a high-fat diet (HFD) to examine its impact. MethodsB16BL6 tumor-bearing mice were fed a HFD. Body weight and food intake were documented every week. We conducted a Western blot analysis to examine the protein levels in the tumor, gastrocnemius (GAS), and tibialis anterior (TA) muscles, as well as the inguinal and epididymal white adipose tissues (iWAT and eWAT). Key findingsEGCG has been shown to have anti-cancer effects equivalent to those of cisplatin, a chemotherapy drug. Furthermore, EGCG protected against the loss of epidydimal white adipose tissue by regulating protein levels of lipolysis factors of adipose triglyceride lipase and hormone-sensitive lipase as well as WAT browning factors of uncoupling protein 1, as opposed to cisplatin. EGCG was shown to reduce the protein levels of muscular atrophy factors of muscle RING-finger protein-1, whereas cisplatin did not contribute to rescuing the atrophy of TA and GAS muscles. ConclusionTaken together, our findings indicate that EGCG has a preventive effect against cachexia symptoms and has anti-cancer effects similar to those of cisplatin in tumor-bearing mice fed a high-fat diet.

In vivo dynamics of hydrogen peroxide and regulation of antioxidant gene expression in mouse skeletal muscle under cooling stimulus

Introduction: The dynamics of reactive oxygen species (ROS) in the cytosol and mitochondria function as signaling factors that regulates diverse cellular adaptations. Hydrogen peroxide (H2O2) is considered one of the most influential ROS in regulating physiological function; being a ubiquitous molecule with a long half-life and high diffusing capacity compared to other ROS (e.g., superoxide and hydroxyl radicals). Temperature changes within skeletal muscle myocytes constitute an environmental factor that modulates the expression of antioxidant-related genes. Although muscle temperature reduction by icing is a typical physical therapy modality, the relationship between cooling stimuli and in vivo H2O2 dynamics remains unclear. The purpose of this investigation was to elucidate the H2O2 dynamics in the cytosol and mitochondria induced by cooling stimuli and to determine the relationship between these dynamics and the expression of antioxidant-related genes. Specifically, we tested the hypothesis that cooling would reduce [H2O2] in intracellular compartments (cytosol and mitochondria) and suppress antioxidant-related gene expression levels. Methods: Experiment 1. in vivo H2O2 dynamics observation: Cytosolic and mitochondrial H2O2-sensitive fluorescent proteins (HyPer7, n = 6 and MLS-HyPer7, n = 9) were expressed by electroporation in the anterior tibialis muscle (TA) of male C57/BL6 mice (9-14 week of age). The TA was exposed under anesthesia, and cytosolic and mitochondrial [H2O2] were measured when the muscle temperature was reduced from 35 °C to 0 °C using in vivo bioimaging techniques. Experiment 2. Mice (9-12 week of age) were divided into an icing (ICE) group that received cooling stimulation (muscle temperature reduced from 35 °C to 13 °C and back to 35°C, total 6 sets for 60 min, n = 6) and a control (CONT) group (muscle temperature 35 °C for 60 min, n = 6). At 3 hours after temperature intervention, the TA was removed and mRNA levels were measured by Real-time PCR. Results: Experiment 1. In vivo [H2O2] in each compartment were significantly lower in the cytosol (over the range 28-6°C) and mitochondria (over the range 29-1°C) compared to 35 °C conditions. A decreasing phase of [H2O2] in both compartments was observed from 35 °C to around 13 °C, but, subsequently increasing [H2O2] were observed in the lower temperature range (i.e., 12 to 0 °C). Experiment 2: The ICE group significantly increased mRNA levels of antioxidant enzyme-related genes Nfe2l2 and HSF1 compared to CONT (1.2 ± 0.1-fold and 1.1 ± 0.1-fold, respectively, P < 0.01). On the other hand, mRNA levels of mitochondria-related genes such as PGC1α and COX4 were unchanged. Conclusions: This investigation is the first to reveal the H2O2 and related gene expression impact of cooling stimuli in an in vivo model. Cooling mouse skeletal muscle produced temperature-dependent decreases in [H2O2] that reversed with more severe cooling. In both cytosolic and mitochondrial compartments, the lowest point for [H2O2] was found to be 13°C. Repeated cooling cycles (between 35 and 13 °C) with their attendant decreasing [H2O2] elevated antioxidant-related mRNA levels. These results support that icing therapy may improve the antioxidant capacity of muscle despite decreasing muscle ROS levels. This study was supported in part by a Grant-in-Aid for Japan Society for the Promotion of Science (JSPS) for Grant-in-Aid for JSPS Fellows (no. JP23KJ0967) and JSPS KAKENHI Grant (No. JP20H04074). 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.

An obesogenic diet alters muscle function in mdx mice

Duchenne muscular dystrophy (DMD) is a progressive muscle degenerative disease that occurs in 1 in 5000 boys born worldwide. DMD is caused by the absence of a functional dystrophin protein, a major structural protein within muscle. Boys with DMD commonly develop obesity and insulin resistance (IR) compounded by limited mobility and extended treatment with glucocorticoids; however, the extent to which obesity and/or IR affect dystrophic pathology are unknown. We hypothesized that a high fat/high sucrose diet (HF/HSD) would further antagonize disease-related losses in muscle function and that these losses would be partially offset by prednisolone (Pred). To test this hypothesis, male C57 and mdx mice were fed a control diet (CD) or HF/HSD for 19 weeks, with or without Pred supplementation, followed by in vitro muscle function tests. Insulin resistance was confirmed via glucose and insulin tolerance tests. The absolute and relative weight of gastrocnemius, soleus, extensor digitorum longus (EDL), and tibialis anterior muscles were increased in mdx groups compared to C57 (p<0.01) but decreased in mdx groups treated with Pred compared to untreated mdx groups (p<0.01). Muscle mass was similar between dietary treatment groups in C57 and mdx mice. Specific tension in the diaphragm was decreased by disease (p<0.01) but was not altered by diet or Pred treatment. Diaphragms from mdx mice were less fatigue resistant than from C57 mice (p<0.01). Within mdx diaphragms, the addition of Pred and/or HF/HSD decreased fatigue resistance compared to mice maintained on CD. The C57 CD/Pred group was less fatigable than other C57 groups (p<0.01). In the soleus, tetanic force was increased in mdx compared to C57 mice (p<0.01) but was decreased by the HF/HSD in mdx groups (p=0.05). Soleus specific tension was similar between C57 and mdx groups but was decreased in mdx HF/HSD groups compared to CD (p<0.01). In the soleus, mdx groups were less resistant to fatigue than C57 groups (p<0.01); however, the HF/HSD and/or Pred treatment did not further promote fatigue in C57 or mdx mice. In EDL, tetanic force was increased in mdx groups compared to C57 groups (p<0.01), and in both C57 and mdx mice, Pred-treated groups had decreased force compared to untreated groups (p<0.01), but force was not impacted by diet. Specific tension was decreased in mdx groups compared to C57 (p<0.01). Within mdx groups, specific tension of Pred-treated groups was increased compared to untreated groups (p<0.01). The EDL was more resistant to contraction-induced injury in C57 compared to mdx mice. Within mdx mice, the HF/HSD and Pred-treated groups were more resistant to eccentric injury than mdx CD/no Pred mice (p<0.01). These data suggest that a HF/HSD and/or Pred can alter parameters of muscle function. This work is supported by Muscular Dystrophy Association grant #962344. 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.

Repair of Sciatic Nerve Defect in Rats With Acellular Nerve Allograft Carrying Vascular Endothelial Cells.

Acellular nerve allografts (ANAs) were developed to replace the autologous nerve grafts (ANGs) to fill the peripheral nerve defects. Poor vascularization relative to ANGs has been a limitation of application of ANAs. A total of 60 female Sprague-Dawley rats were assigned 3 groups. The rats in A group received ANGs, the rats in B group received ANAs, and the rats in C group were transplanted with ANA carrying endothelial cells (ANA + ECs). In the 1st, 2nd, 4th, and 12th postoperative weeks, 5 rats were selected from each group for evaluating sciatic function index (SFI), electrophysiology, maximum tetanic force recovery rate, tibialis anterior muscle weights recovery rate, and microvessel density. In the 12th postoperative week, the nerves were harvested and stained with toluidine blue and observed under an electron microscope to compare nerve fibers, myelin width, and G-ratio. All the rats survived. In the first and second postoperative weeks, more microvessels were found in the ANA + EC group. In the 12th postoperative week, the nerve fibers were more numerous, and G-ratio was smaller in the C group compared with the B group. The compound muscle action potential and maximum tetanic force recovery rate in the tibialis anterior muscle in the C group were better than those in the B group in the 12th postoperative week. The A group showed better performances in electrophysiology, maximum tetanic force, muscle wet weight, and nerve regeneration. ANA + ECs can promote early angiogenesis, promoting nerve regeneration and neurological function recovery.

Maintenance of muscle fiber-type memory in a human-to-mouse muscle xenograft model

INTRODUCTION. Muscle tissue can regenerate through muscle stem cells known as muscle satellite cells (MuSCs). MIME is a technique developed in our laboratory, through which, donor muscle tissue with MuSCs can be implanted into a host muscle to promote donor-cell-derived myogenesis. OBJECTIVE. We studied whether or not MuSCs maintain memory of the type of muscle fibers (fast or slow) with which they were associated. We hypothesized that MuSCs do not maintain memory of the fibers with which they are associated and form slow or fast muscle fibers based on the host environment. METHODS. We tested our hypothesis by implanting donor muscle, which contained mostly slow fibers, into host muscle, which contained mostly fast fibers. Implantation of donor tissue was performed by MIME. Following MIME, we injected a myotoxin (1.2% barium chloride, BaCl2) into the host muscle, which induced concerted degeneration followed by MuSC-dependent regeneration. Specifically, we studied mouse tibialis anterior muscles, which were implanted with segments of human donor tibialis anterior muscle. The human TA is comprised mostly of slow fibers (~60%; i.e., slow myosin heavy chain positive [sMyHC+]). The mouse TA is comprised 100% of fast fibers (i.e., fast myosin heavy chain positive [fMyHC+] fibers). This experimental paradigm is appropriate to ascertain whether or not satellite cells retain memory of the muscle fibers with which they were originally associated. We collected host muscles after 12 weeks of regeneration to ascertain if there was an increase in the percentage of slow muscle fibers. We used antibodies specific to sMyHC and fMyHC to stain frozen TA muscle sections. Host mice were immunodeficient and ubiquitously expressed enhanced green fluorescent protein (NSG-EGFP, The Jackson Laboratory Strain #021937). Since host mouse muscle fibers exhibited green fluorescence, but donor human muscle fibers did not, host and donor muscle fibers could be easily differentiated under fluorescence optics. RESULTS. Quantitative data confirmed that the cluster of human muscle fibers, which regenerated in the host mouse TA muscle had more slow (~50%) than fast (~30%) fibers (T-test, p<0.01, N = 4 TA muscles from 4 different host mice). CONCLUSION. Contrary to our hypothesis, our data suggest that MuSCs do maintain memory of the type of muscle fibers with which they were originally associated. MuSCs give rise to fast or slow muscle fibers based on their donor origin when they are implanted along with their original donor muscle tissue. This work was supported by NIH R03HD091648, Pitch Award from the Alliance for Regenerative Rehabilitation Research and Training (AR3T) from NIH P2CHD086843, and a subcontract from NIH R01AR079884-01 (Peter L. Jones PI) to JAR. We thank the Wayne State University DPT Program for supporting student research and providing research training through coursework and research experiences. 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.

Muscle regeneration is not impaired by ablation of UCP-1-expressing cells

INTRODUCTION: Recently, cells expressing uncoupling protein 1 (UCP-1) have been proposed to impact muscle physiology. First, transplant of brown adipose tissue (which has high expression of UCP-1) enhanced regeneration in the injured mouse rotator cuff. Second, isolated UCP-1+ muscle fibroadipogenic progenitors (FAPs) treated to enhance UCP-1 expression improved injured mouse rotator cuff muscle pathology. However, it remains unknown whether endogenous UCP-1-expressing cells contribute to muscle regeneration. Therefore, the objective of this study was to assess whether mice that lack UCP-1-expressing cells demonstrate impaired muscle regeneration following toxin administration. METHODS: Experiments were performed using tibialis anterior (TA) and 5th toe extensor digitorum longus (EDL) muscle of wild type C57BL/6J (WT; n=22), and uncoupling protein 1 diphtheria Toxin A (UCP1-DTA; n=18) mice, which lack all UCP-1 expressing cells. All procedures were performed in accordance with the National Institute of Health’s Guide for the Use and Care of Laboratory Animals and were approved by Washington University School of Medicine IACUC. Muscle injury was induced using intramuscular injection of 50%v/v solution of glycerol. Regeneration was assessed at “early” (7 days post-injury (dpi)), “intermediate” (14 dpi) and “late” (21 dpi) stages. Outcome measures were cell population dynamics by flow cytometry, ex-vivo physiology testing and histological scoring as appropriate for each timepoint. RESULTS: Our preliminary data in early regeneration show that UCP1-DTA mice have significantly greater percentage of FAPs compared to WT mice (29.84 ± 1.24% higher cell population, p<0.0134), however no differences in muscle satellite (stem) cells between genotypes. Physiological and histological outcomes at intermediate and late-stage regeneration find a baseline deficit in muscle mass (23.69 ± 2.3% smaller, p<0.0001), type 2B fiber cross-sectional area (32.56 ± 4.8% smaller, p<0.0001) and normalized muscle active force generation (16.2 ± 3.3% lower force, p<0.0154) in UCP1-DTA compared with WT. However, mass and contractile forces recover to a similar extent between the genotypes following glycerol injection suggesting the phenotypic drivers of the reduced mass and contractile function do not affect muscle regeneration. CONCLUSIONS: Taken together, the data from this study point to a baseline growth deficit in mice lacking UCP-1-expressing cells. However, in two separate timepoints of regeneration (14 and 21dpi), we see that UCP1-DTA mice have similar recovery of mass and normalized contractile force to baseline values suggesting regeneration can proceed normally without UCP-1 expressing cells. However, further data collection will be needed at early-stage regeneration to help discern whether mice lacking UCP-1-expressing cells have initial changes in cell population dynamics given the increased FAP population at 7dpi. ACKNOWLEDGMENTS: This work was supported by R01AR075773 and R21AR071582 to GAM. 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.

Loss of FAM210A in Satellite Cell Drives Mitochondrial Dysfunction and Muscle Weakness in Experimental Peripheral Artery Disease

Introduction: Peripheral artery disease (PAD) affects approximately 200 million people globally. Chronic limb-threatening ischemia (CLTI) represents the most advanced clinical phenotype of PAD. Mitochondrial metabolism defects are apparent in the satellite cells of CLTI patients, and the muscle regeneration process demands increased energy, with satellite cell mitochondrial metabolism and dynamics playing a crucial role. However, our current understanding of the role of satellite cells and muscle regeneration in PAD pathology is notably inadequate. Recent studies have shown that the absence of FAM210A, a newly identified mitochondrial protein, can trigger morphological and energetic abnormalities in mitochondria, resulting in muscle function deterioration and promoting muscle wasting. Given the impaired mitochondrial function in satellite cells in CLTI patients and the heightened energy demand regulated by mitochondrial dynamics during muscle regeneration in satellite cell, it is therefore essential to exploring the role of FAM210A in satellite cells in CLTI limb pathology. The objective of this study was to investigate the impact of the loss of FAM210A in satellite cells in mice with experimental PAD. Methods: Male and female conditional FAM210A satellite cell knock out (FAM210ASCKO) and FAM210A floxed (FAM210Afl/fl) mice (N=3-5) were used for this project. Femoral artery ligation surgery was performed to induce PAD. Tamoxifen was administered for 5 days intraperitoneally one week prior to the surgery. Mice were euthanized twenty-eight days after surgery. Outcome measures included limb perfusion assessment, muscle weight, muscle force production, and mitochondrial respiration. Results: FAM210ASCKO mice had significantly decreased muscle mass of the tibialis anterior and extensor digitorum longus ( p<0.0001 and p=0.0004 respectively). More importantly, maximal muscle force with in-situ contraction in tibialis anterior was significantly lower in FAM210ASCKO mice compared to FAM210Afl/fl mice (FAM210Afl/fl mice: 1085.16 ± 331.08 mN; FAM210ASCKO mice: 430.77 ± 467.91 mN; p=0.0139). Furthermore, analysis of mitochondrial oxidative phosphorylation (OXPHOS) revealed that FAM210ASCKO mice had a significant reduction compared to FAM210Afl/fl mice (48%) in skeletal muscle mitochondrial bioenergetic function at various levels of energy demand mimicking from resting to maximal exercise when carbohydrate was used to energize mitochondria ( p=0.034). Importantly, the analysis of mitochondrial membrane potential consistently reveals a substantial reduction in FAM210ASCKO mice compared to FAM210Afl/fl mice, which can partially explain the diminished mitochondrial bioenergetic function observed in ischemic FAM210ASCKO mice. Conclusion: The results of this study indicate that the loss of FAM210A in satellite cells exacerbates ischemic muscle pathology evidenced by muscle weakness and mitochondrial failure in FAM210ASCKO mice with PAD. Future work is needed to elucidate the mechanisms by which FAM210A regulate ischemic satellite cell function and the therapeutic potential of preventing ischemia-induced loss of FAM210A in PAD. This study was supported by National Institutes of Health (NIH) grant R01-HL149704 (to Dr Ryan). Mr Thome was supported by NIH grant F31-DK128920. 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.

Surface electromyography analysis of ankle flexor and extensor activity under different standing stability levels.

Background: To observe the activation strategies of the ankle muscles using surface electromyography (sEMG) during single-leg standing (SLS) and both-leg standing (BLS) on flat ground (FG), soft mat (SM), and BOSU ball (BB) surfaces. Methods: Thirty healthy young adults participated in the study. The muscle activities of the tibialis anterior (TA) and gastrocnemius medial (GM) were measured on the three surfaces during SLS and BLS. Electromyographic evaluations of the TA and GM were recorded during maximum voluntary isometric contractions (MVIC). Muscle activation was evaluated using MVIC%, and muscle co-contraction was evaluated using the co-contraction index (CI). Results: A statistically significant increase was observed in the MVIC% of the TA, GM, and CI on the three surfaces during SLS compared to BLS, except for the comparison of CI on BB between SLS and BLS (t = -1.35, p = 0.19). The MVIC% of the TA and GM during SLS and BLS on BB was significantly increased in comparison with FG and SM. The CI during BLS on BB increased compared to FG (t = 3.19, p < 0.01) and SM (t = 4.64, p < 0.01). The CI during BLS on SM (t = -1.46, p = 0.15) decreased when compared to FG but without statistical significance. Conclusions: SLS and unstable surfaces can induce greater muscle activation, and SLS can have a greater influence on ankle muscles.

Captopril treatment does not affect inflammatory and autophagy signaling in skeletal muscle of Zucker diabetic fatty rat

Skeletal muscle plays a key role in the development of Type 2 Diabetes Mellitus (T2DM) as its large capacity to sequester glucose and contribute to the regulation of energy metabolism is compromised. Previous studies indicate that in skeletal muscle, T2DM drives inflammatory signaling and impairs autophagy. Captopril (Cap), an angiotensin-converting enzyme (ACE) inhibitor, is a common treatment for hypertension in T2DM patients; however, the impact of Cap on inflammatory signaling and autophagy in insulin resistant skeletal muscle remains ill-defined. We hypothesized that inflammatory signaling would be elevated and degradation of autophagosomes would be impaired in obese Zucker diabetic fatty (ZDF) rats, which are hyperphagic as they lack the leptin receptor. We further hypothesized that daily Cap treatment would attenuate these disturbances. At 14 weeks of age, the absolute weights of gastrocnemius (p&lt;0.0001), tibialis anterior (p&lt;0.0001), soleus (p=0.002), and extensor digitorum longus (EDL, p&lt;0.0001) muscles were decreased by 23-34% in ZDF compared to lean rats but were not altered by 8 weeks of Cap treatment (0.5g/kg/day). Immunoblotting revealed that in soleus and EDL, inflammatory signaling proteins TLR4, phosphorylated (p)IKK α/β (ser176/180), pNFkB p65 (Ser 536), IkBα, TNFα, and AP1 were similar amongst all groups, whereas MyD88 was elevated in ZDF soleus (p=0.03) but was not corrected by Cap. The autophagy protein, LC3A/B I was increased 32% (p=0.01) in EDL from ZDF rats compared to Con; however, PI3K III, pBeclin1 (ser 93), pULK1 (ser555), LAMP2, LC3A/B II, and the LC3A/B II/I ratio were similar amongst groups in the soleus and EDL and were not altered by Cap. Compared to control, P62, a key autophagy adapter protein, was similar in soleus and decreased 27% in EDL (p&lt;0.01) from ZDF rats and was not altered by Cap. Counter to our expectations, although muscle mass was lower in ZDF groups, obesity and insulin resistance in young ZDF rats were not suffcient to drive inflammatory signaling or disrupt autophagy in glycolytic or oxidative skeletal muscle, and these processes appeared to be resistant to Cap treatment in ZDF rats. USDA-NIFA awarded to MJR (2021-67017-33937). 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.

Novel Method to Assess Treatment Effcacy in Preclinical Peripheral Arterial Disease Studies

Background: Peripheral arterial disease (PAD), a condition caused by atherosclerotic narrowing or occlusion of blood vessels in the lower extremities, affects ~200 million adults globally and lacks effective treatment options to improve limb function. Cilostazol is the only approved pharmacological treatment option for PAD patients, but its effect on walking performance is modest and the drug is not widely used clinically. Over the past several decades, numerous therapies (drug, gene, and cell-based) have shown promise in the preclinical setting but failed to achieve primary outcomes in clinical trials. While the failure of therapeutic translation is complex, most preclinical studies have employed perfusion recovery as a primary outcome variable whereas most clinical trials in PAD patients employ the six-minute walk test. Although perfusion is an important contributor to limb function and walking performance, assessing preclinical therapeutics solely via perfusion recovery may be a flawed approach. Objective: To address the need for a more comprehensive method to assess the effcacy of preclinical PAD therapeutics, we aimed to develop a protocol to assess hindlimb function that incorporates assessment of muscle power and perfusion across a six-minute testing period. Methods: To assess muscle contractile performance, the tibialis anterior or plantarflexor muscles were carefully exposed and the distal tendon mounted to the force-length controller (Aurora Scientific) and the muscles were stimulated via the sciatic nerve in anesthetized C57BL6 mice. After obtaining isometric forces, a six-minute functional test was performed using repeated isotonic (shortening) contractions against a submaximal load (30% or 45% of the reference force). Muscle fatigue was measured as a change in power across the testing period. Throughout the test, laser Doppler flowmetry was used to measure muscle blood flow. Hindlimb ischemia was used as an experimental model of PAD. Results: In healthy mice without PAD, the test produced the expected dose-dependent relationship between fatigue and workload with contractions performed at 30% of the reference force having less fatigue compared to 45% of the reference force ( P=0.0051), without a significant difference in muscle blood flow between the conditions. Compared the mice that received a sham surgery, those subjected to hindlimb ischemia displayed significantly lower maximal isometric forces (1514 ± 118 mN vs. 880 ± 85 mN, P=0.0001) and power at 35% maximal force (280 ± 12 vs. 105 ± 46 watts/kg, P=0.0002). Whereas sham mice lost ~25% power across the six-minute test, HLI mice displayed a ~70% loss of power. Regarding muscle blood flow, sham mice increased blood flow ~250% whereas HLI mice were only able to increase blood flow to ~105% ( P&lt;0.001). Conclusions: This newly developed protocol successfully combines isotonic muscle function and limb perfusion assessments in the hindlimb of mice and can detect differences between healthy and PAD mice. Future work is needed to fully evaluate the accuracy and sensitivity of this method to select therapeutics in preclinical PAD models. This study was supported by National Institutes of Health (NIH) grant R01-HL149704 (T.E.R.). 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.

Tibiofemoral bone configuration is not associated with hamstring muscle strength in male and female patients with ACL reconstruction.

Previous evidence indicated that the tibiofemoral bone configuration might elevate the risk of an anterior cruciate ligament (ACL) injury. Furthermore, a low hamstring-to-quadriceps muscle ratio predisposes especially females to unfavourable knee kinematics. The primary objective of the present study was to investigate sex-specific associations between tibiofemoral bone geometry and isokinetic knee flexion torque in patients with primary ACL injury followed by ACL reconstruction. N = 100 patients (72 = male, 28 = female, age = 31.3 ± 10.2, body mass index = 25.3 ± 3.6) with primary ACL rupture with isokinetic knee flexion torque assessments before and 6 months after ACL reconstruction surgery were analysed. Magnetic resonance imaging scans were analysed for medial posterior tibial slope (MPTS) and lateral posterior tibial slope, notch width index (NWI) and lateral femoral condyle index (LFCI). Additionally, isokinetic knee flexion torque (60°/s) and hamstring-quadriceps ratios were evaluated. Subsequently, functional parameters were correlated with imaging data for gender subgroups. The findings showed that presurgical isokinetic knee flexion torque was not associated with any marker of femoral or tibial bone geometry. Further, while significant differences were observed between female (0.883 ± 0.31 Nm/kg) and male (1.18 ± 0.35 Nm/kg) patients regarding preoperative normalized knee flexion torque (p < 0.001), no significant sex differences were found for percentage increases in normalized knee flexion torque from presurgery to postsurgery. Generally, female patients demonstrated significantly higher MPTS magnitudes (p < 0.05) and lower LFCI values (p < 0.05) compared to men. The present results demonstrated no association between tibial or femoral bone geometry and muscle strength of the hamstrings in patients with ACL reconstruction, indicating an important mismatch of muscular compensation to deviations in bone geometry. There were no sex-specific differences in tibiofemoral bone parameters. Level III.

POSTURAL CONTROL IMPAIRED IN MYOARTHROKINETIC AND NEURO-SENSORY DISORDERS

Interdependent subsystems involved in dynamic posture and locomotion are represented by osteoarticular, neuromuscular, sensory systems. Postural control is maintained by somato-sensory, visual vestibular feedback, integrated at the level of the locomotor system and the central nervous system. Postural stability is disturbed in mio-arthro-kinetic disorders. In proprioceptive loss, the automatic postural responses are diminished in amplitude and delayed in time; stretch reflexes in soleus and medial gastrocnemius are severely diminished; balance-correcting responses in tibialis anterior are diminished and delayed; backward motion of the trunk is reduced; movement strategies and synergies are disturbed. In peripheral vestibular loss, there are increased trunk responses to disruptive stimuli, with hypermetric trunk muscle responses and hypometric knee responses, but unchanged synergies; pathological changes to the modulation depth are produced; the amplitude of balance correcting responses in ankle muscles is severely reduced. Visual input serves to decrease the stiffness of the musculoskeletal system; it can be accomplished by decreasing the level of muscular activity across the joints of the lower limbs (in eyes-open subjects) or by reducing the gains of the other postural feedback mechanisms, the proprioceptive or vestibular systems (in eyes-closed subjects). Keywords: postural control, proprioceptive insufficiency, vestibular impairment.

Spectral characterization of human leg EMG signals from an open access dataset for the development of computational models.

Large-scale neuromusculoskeletal models have been used for predicting mechanisms underlying neuromuscular functions in humans. Simulations of such models provide several types of signals of practical interest, such as surface electromyographic signals (EMG), which are compared with experimental data for interpretations of neurophysiological phenomena under study. Specifically, realistic characterization of spectral properties of simulated EMG signals is important for achieving powerful inferences, whereas considerations should be taken for myoelectric signals of different muscles. In this study, we characterized spectral properties of surface interference pattern EMG signals and motor unit action potentials (MUAP) acquired from three plantar flexor muscles: Soleus (SO), Medial Gastrocnemius (MG), and Lateral Gastrocnemius (LG); and one dorsiflexor muscle: Tibialis Anterior (TA). Surface EMG signals were acquired from 20 participants using the same convention for electrode placement. Specifically, interference pattern EMG signals were obtained during isometric constant force contractions at 5%, 10% and 20% of maximum voluntary contraction (MVC), whereas surface MUAPs were decomposed from surface EMG signals obtained at low contraction forces. We compared the spectrum median frequency (MDF) estimated from interference pattern EMG signals across muscles and contraction intensities. Additionally, we compared MDF and durations of MUAPs between muscles. Our results showed that MDF of interference pattern EMG signals acquired from TA were higher compared to SO, MG, and LG for all contraction intensities i.e., 5%, 10%, and 20% MVC. Consistently, MUAPs acquired from TA also had higher MDF values and shorter durations compared to the other leg muscles. We provide herein a dataset with the surface MUAPs waveforms and interference pattern EMG signals obtained for this study, which should be useful for implementing and validating the simulation of myoelectrical signals of leg muscles. Importantly, these results indicate that spectral properties of myoelectrical signals should be considered for improving EMG modeling in large-scale neuromusculoskeletal models.

UBE2C promotes myoblast differentiation and skeletal muscle regeneration through the Akt signaling pathway.

Ubiquitin-conjugation enzyme E2C (UBE2C) is a crucial component of the ubiquitin-proteasome system that is involved in numerous cancers. In this study, we find that UBE2C expression is significantly increased in mouse embryos, a critical stage during skeletal muscle development. We further investigate the function of UBE2C in myogenesis. Knockdown of UBE2C inhibits C2C12 cell differentiation and decreases the expressions of MyoG and MyHC, while overexpression of UBE2C promotes C2C12 cell differentiation. Additionally, knockdown of UBE2C, specifically in the tibialis anterior muscle (TA), severely impedes muscle regeneration in vivo. Mechanistically, we show that UBE2C knockdown reduces the level of phosphorylated protein kinase B (p-Akt) and promotes the degradation of Akt. These findings suggest that UBE2C plays a critical role in myoblast differentiation and muscle regeneration and that UBE2C regulates myogenesis through the Akt signaling pathway.

A comprehensive dataset on biomechanics and motor control during human walking with discrete mechanical perturbations.

Humans have a remarkable capability to maintain balance while walking. There is, however, a lack of publicly available research data on reactive responses to destabilizing perturbations during gait. Here, we share a comprehensive dataset collected from 10 participants who experienced random perturbations while walking on an instrumented treadmill. Each participant performed six 5-min walking trials at a rate of 1.2 m/s, during which rapid belt speed perturbations could occur during the participant's stance phase. Each gait cycle had a 17% probability of being perturbed. The perturbations consisted of an increase of belt speed by 0.75 m/s, delivered with equal probability at 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 80% of the stance phase. Data were recorded using motion capture with 25 markers, eight inertial measurement units (IMUs), and electromyography (EMG) from the tibialis anterior (TA), soleus (SOL), lateral gastrocnemius (LG), rectus femoris (RF), vastus lateralis (VL), vastus medialis (VM), biceps femoris (BF), and gluteus maximus (GM). The full protocol is described in detail. We provide marker trajectories, force plate data, EMG data, and belt speed information for all trials and participants. IMU data is provided for most participants. This data can be useful for identifying neural feedback control in human gait, biologically inspired control systems for robots, and the development of clinical applications.

Concerted regulation of skeletal muscle metabolism and contractile properties by the orphan nuclear receptor Nr2f6.

The maintenance of skeletal muscle plasticity upon changes in the environment, nutrient supply, and exercise depends on regulatory mechanisms that couple structural and metabolic adaptations. The mechanisms that interconnect both processes at the transcriptional level remain underexplored. Nr2f6, a nuclear receptor, regulates metabolism and cell differentiation in peripheral tissues. However, its role in the skeletal muscle is still elusive. Here, we aimed to investigate the effects of Nr2f6 modulation on muscle biology in vivo and in vitro. Global RNA-seq was performed in Nr2f6 knockdown C2C12 myocytes (N=4-5). Molecular and metabolic assays and proliferation experiments were performed using stable Nr2f6 knockdown and Nr2f6 overexpression C2C12 cell lines (N=3-6). Nr2f6 content was evaluated in lipid overload models in vitro and in vivo (N=3-6). In vivo experiments included Nr2f6 overexpression in mouse tibialis anterior muscle, followed by gene array transcriptomics and molecular assays (N=4), ex vivo contractility experiments (N=5), and histological analysis (N=7). The conservation of Nr2f6 depletion effects was confirmed in primary skeletal muscle cells of humans and mice. Nr2f6 knockdown upregulated genes associated with muscle differentiation, metabolism, and contraction, while cell cycle-related genes were downregulated. In human skeletal muscle cells, Nr2f6 knockdown significantly increased the expression of myosin heavy chain genes (two-fold to three-fold) and siRNA-mediated depletion of Nr2f6 increased maximal C2C12 myocyte's lipid oxidative capacity by 75% and protected against lipid-induced cell death. Nr2f6 content decreased by 40% in lipid-overloaded myotubes and by 50% in the skeletal muscle of mice fed a high-fat diet. Nr2f6 overexpression in mice resulted in an atrophic and hypoplastic state, characterized by a significant reduction in muscle mass (15%) and myofibre content (18%), followed by an impairment (50%) in force production. These functional phenotypes were accompanied by the establishment of an inflammation-like molecular signature and a decrease in the expression of genes involved in muscle contractility and oxidative metabolism, which was associated with the repression of the uncoupling protein 3 (20%) and PGC-1α (30%) promoters activity following Nr2f6 overexpression in vitro. Additionally, Nr2f6 regulated core components of the cell division machinery, effectively decoupling muscle cell proliferation from differentiation. Our findings reveal a novel role for Nr2f6 as a molecular transducer that plays a crucial role in maintaining the balance between skeletal muscle contractile function and oxidative capacity. These results have significant implications for the development of potential therapeutic strategies for metabolic diseases and myopathies.