Fiber Type Distribution Research Articles

Effects of gene replacement therapy with resamirigene bilparvovec (AT132) on skeletal muscle pathology in X-linked myotubular myopathy: results from a substudy of the ASPIRO open-label clinical trial

X-linked myotubular myopathy (XLMTM) is a rare, life-threatening congenital muscle disease caused by mutations in the MTM1 gene that result in profound muscle weakness, significant respiratory insufficiency, and high infant mortality. There is no approved disease-modifying therapy for XLMTM. Resamirigene bilparvovec (AT132; rAAV8-Des-hMTM1) is an investigational adeno-associated virus (AAV8)-mediated gene replacement therapy designed to deliver MTM1 to skeletal muscle cells and achieve long-term correction of XLMTM-related muscle pathology. The clinical trial ASPIRO (NCT03199469) investigating resamirigene bilparvovec in XLMTM is currently paused while the risk:benefit balance associated with this gene therapy is further investigated. Muscle biopsies were taken before treatment and 24 and 48 weeks after treatment from ten boys with XLMTM in a clinical trial of resamirigene bilparvovec (ASPIRO; NCT03199469). Comprehensive histopathological analysis was performed. Baseline biopsies uniformly showed findings characteristic of XLMTM, including small myofibres, increased internal or central nucleation, and central aggregates of organelles. Biopsies taken at 24 weeks post-treatment showed marked improvement of organelle localisation, without apparent increases in myofibre size in most participants. Biopsies taken at 48 weeks, however, did show statistically significant increases in myofibre size in all nine biopsies evaluated at this timepoint. Histopathological endpoints that did not demonstrate statistically significant changes with treatment included the degree of internal/central nucleation, numbers of triad structures, fibre type distributions, and numbers of satellite cells. Limited (predominantly mild) treatment-associated inflammatory changes were seen in biopsy specimens from five participants. Muscle biopsies from individuals with XLMTM treated with resamirigene bilparvovec display statistically significant improvement in organelle localisation and myofibre size during a period of substantial improvements in muscle strength and respiratory function. This study identifies valuable histological endpoints for tracking treatment-related gains with resamirigene bilparvovec, as well as endpoints that did not show strong correlation with clinical improvement in this human study. Astellas Gene Therapies (formerly Audentes Therapeutics, Inc.).

The impact of life-long strength versus endurance training on muscle fiber morphology and phenotype composition in older men.

Aging is typically associated with decreased muscle strength and rate of force development (RFD), partly explained by motor unit remodeling due to denervation, and subsequent loss of fast-twitch type II myofibers. Exercise is commonly advocated to counteract this detrimental loss. However, it is unclear how life-long strength versus endurance training may differentially affect markers of denervation and reinnervation of skeletal myofibers and, in turn, affect the proportion and morphology of fast-twitch type II musculature. Thus, we compared fiber type distribution, fiber type grouping, and the prevalence of atrophic myofibers (≤1,494 µm2) in strength-trained (OS) versus endurance-trained (OE) master athletes and compared the results to recreationally active older adults (all >70 yr, OC) and young habitually active references (<30 yr, YC). Immunofluorescent stainings were performed on biopsy samples from vastus lateralis, along with leg press maximal strength and RFD measurements. OS demonstrated similar type II fiber distribution (OS: 52.0 ± 16.4%; YC: 51.1 ± 14.4%), fiber type grouping, maximal strength (OS: 170.0 ± 18.9 kg, YC: 151.0 ± 24.4 kg), and RFD (OS: 3,993 ± 894 N·s-1, YC: 3,470 ± 1,394 N·s-1) as young, and absence of atrophic myofibers (OS: 0.2 ± 0.7%; YC: 0.1 ± 0.4%). In contrast, OE and OC exhibited more atrophic fibers (OE: 1.2 ± 1.0%; OC: 1.1 ± 1.4%), more grouped fibers, and smaller proportion of type II fibers (OE: 39.3 ± 11.9%; OC: 35.0 ± 12.4%) than OS and YC (all P < 0.05). In conclusion, strength-trained master athletes were characterized by similar muscle morphology as young, which was not the case for recreationally active or endurance-trained old. These results indicate that strength training may preserve type II fibers with advancing age in older men, likely as a result of chronic use of high contractile force generation.NEW & NOTEWORTHY Aging is associated with loss of fast-twitch type II myofibers, motor unit remodeling, and grouping of myofibers. This study reveals, for the first time, that strength training preserves neural innervation of type II fibers, resulting in similar myofiber type distribution and grouping in life-long strength-trained master athletes as young moderately active adults. In contrast, life-long endurance-trained master athletes and recreationally active old adults demonstrated higher proportion of type I fibers accompanied by more marked grouping of type I myofibers, and more atrophic fibers compared with strength-trained master athletes and young individuals. Thus, strength training should be utilized as a training modality for preservation of fast-twitch musculature, maximal muscle strength, and rapid force capacity (RFD) with advancing age.

Inhaled beta2 -agonist, formoterol, enhances intense exercise performance, and sprint ability in elite cyclists.

Many athletes use long-acting beta2 -agonist formoterol in treatment of asthma. However, studies in non-athlete cohorts demonstrate that inhaled formoterol can enhance sprint performance calling into question whether its use in competitive sports should be restricted. We investigated whether formoterol at upper recommended inhaled doses (54 μg) would enhance sprint ability and intense exercise performance in elite cyclists. Twenty-one male cyclists (V̇O2max : 70.4 ± 4.3 mL × min-1 × kg-1 , mean ± SD) completed two 6-s all-out sprints followed by 4-min all-out cycling after inhaling either 54 μg formoterol or placebo. We also assessed cyclists' leg muscle mass by dual-energy X-ray absorptiometry and muscle fiber type distribution of vastus lateralis biopsies. Peak and mean power output during the 6-s sprint was 32 W (95% CI, 19-44 W, p < 0.001) and 36 W (95% CI, 24-48 W, p < 0.001) higher with formoterol than placebo, corresponding to an enhancing effect of around 3%. Power output during 4-min all-out cycling was 9 W (95% CI, 2-16 W, p = 0.01) greater with formoterol than placebo, corresponding to an enhancing effect of 2.3%. Performance changes in response to formoterol were unrelated to cyclists' VO2max and leg lean mass, whereas muscle fiber Type I distribution correlated with change in sprinting peak power in response to formoterol (r2 = 0.314, p = 0.012). Our findings demonstrate that an inhaled one-off dose of 54 μg formoterol has a performance-enhancing potential on sprint ability and short intense performance in elite male cyclists, which is irrespective of training status but partly related to muscle fiber type distribution for sprint ability.

The effects of environmental enrichment on voluntary physical activity and muscle mass gain in growing rats.

Introduction: Environmental enrichment (EE) for rodents involves housing conditions that facilitate enhanced sensory, cognitive, and motor stimulation relative to standard housing conditions. A recent study suggested that EE induces muscle hypertrophy. However, it remains unclear whether muscle hypertrophy in EE is associated with voluntary physical activity, and the characteristics of muscle adaptation to EE remain unclarified. Therefore, this study investigated whether muscle adaptation to EE is associated with voluntary physical activity, and assessed the changes in the muscle fiber-type distribution and fiber-type-specific cross-sectional area in response to EE. Methods: Wistar rats (6weeks of age) were randomly assigned to either the standard environment group (n = 10) or the EE group (n = 10). The voluntary physical activity of rats housed in EE conditions was measured using a recently developed three-axis accelerometer. After exposure to the standard or enriched environment for 30days, the tibialis anterior, extensor digitorum longus, soleus, plantaris, and gastrocnemius muscles were removed and weighed. Immunohistochemistry analysis was performed on the surface (anterior) and deep (posterior) areas of the tibialis anterior and soleus muscles. Results and discussion: The EE group showed increased voluntary physical activity during the dark period compared with the standard environment group (p = 0.005). EE induced muscle mass gain in the soleus muscle (p = 0.002) and increased the slow-twitch muscle fiber cross-sectional area of the soleus muscle (p = 0.025). EE also increased the distribution of high-oxidative type IIa fibers of the surface area (p = 0.001) and type I fibers of the deep area (p = 0.037) of the tibialis anterior muscle. These findings suggest that EE is an effective approach to induce slow-twitch muscle fiber hypertrophy through increased daily voluntary physical activity.

Differences in Skeletal Muscle Fiber Characteristics Between Affected and Nonaffected Limbs in Individuals With Stroke: A Scoping Review.

The objective of this scoping review was to characterize and identify knowledge gaps about the changes in skeletal muscle fiber type proportion and cross-sectional area (CSA) after stroke. This scoping review followed previously proposed frameworks. A systematic search was conducted for articles examining muscle fiber type proportion and CSA in individuals with stroke in EMBASE, MEDLINE, PsycINFO, CINAHL, SPORTDiscus, and Web of Science databases from inception to December 20, 2022. Two independent authors screened and extracted the data. Results were discussed using theories proposed by the authors of the included studies. Of 13 studies (115 participants), 6 (46%) were case studies or case series, 6 (46%) were cross-sectional studies, and 1 (8%) was an experimental study. Studies had small sample sizes (1-23 participants) and various muscle sampling sites (6 different muscles). All 13 studies examined muscle fiber type distributions, and 6 (46%) examined CSA. Ten (77%) studies examined differences between paretic and nonparetic muscles, and 5 (38%) compared people with stroke to people without stroke. Results from 9 of 13 studies (69%) supported a greater proportion of type II muscle fibers in the paretic limb. Of those, 4 studies (42 participants), 3 studies (17 participants), and 1 study (1 participant) saw no differences, preferential type II and type I CSA loss between limbs, respectively. Of the limited available evidence, stroke appears to result in a shift to a higher proportion of type II muscle fibers in the paretic muscles. There are mixed results for effects on muscle fiber CSA, but there is some evidence of specific atrophy of type II muscle fibers. Changes in paretic skeletal muscle fibers of individuals with stroke may explain, in part, the substantial losses in strength and power in this population. Interventions to restore type II muscle fiber size may benefit people with stroke.

Myosin Heavy-Chain Messenger Ribonucleic Acid (mRNA) Expression and Fibre Cross-Sectional Area in Masseter, Digastric, Gastrocnemius and Soleus Muscles of Young and Adult Rats.

Different demands on the muscles of mastication may influence their functional profile (size and distribution of muscle fibre types), which may change during growth and maturation, potentially influencing craniofacial growth. The aim of this study was to evaluate mRNA expression and cross-sectional area of masticatory muscle fibres compared with limb muscles in young and adult rats. Twenty-four rats were sacrificed at two different ages, namely 12 at 4 weeks (young) and 12 at 26 weeks (adult). The masseter, digastric, gastrocnemius and soleus muscles were dissected. Gene expression of myosin heavy-chain isoforms Myh7 (MyHC-I), Myh2 (MyHC-IIa), Myh4 (MyHC-IIb) and Myh1 (MyHC-IIx) in the muscles was measured using qRT-PCR RNA analysis, and immunofluorescence staining was performed to measure the cross-sectional area of different muscle fibre types. Different muscle types and ages were compared. Significant differences were found in the functional profile between masticatory and limb muscles. For the masticatory muscles, there was an increase in Myh4 expression with age, and this change was more intense for the masseter muscles, which also presented an increase in Myh1 expression, similarly to limb muscles. The fibre cross-sectional area of the masticatory muscles was generally smaller in young rats; however, this difference was less pronounced than in limb muscles.

Skeletal muscle fibre type and enzymatic activity in adult offspring following placental and peripheral malaria exposure in foetal life.

Maternal malaria may restrict foetal growth. Impaired utero-placental blood flow due to malaria infection may cause hypoxia-induced altered skeletal muscle fibre type distribution in the offspring, which may contribute to insulin resistance and impaired glucose metabolism. This study assessed muscle fibre distribution 20 years after placental and/or peripheral in-utero malaria exposure compared to no exposure, i.e., PPM+, PM+, and M-, respectively. We traced 101 men and women offspring of mothers who participated in a malaria chemosuppression study in Muheza, Tanzania. Of 76 eligible participants, 50 individuals (29 men and 21 women) had skeletal muscle biopsy taken from m. vastus lateralis in the right leg. As previously reported, fasting and 30 min post-oral glucose challenge plasma glucose values were higher, and insulin secretion disposition index was lower, in the PPM+ group. Aerobic capacity (fitness) was estimated by an indirect VO2max test on a stationary bicycle. Muscle fibre sub-type (myosin heavy chain, MHC) distribution was analysed, as were muscle enzyme activities (citrate synthase (CS), 3-hydroxyacyl-CoA dehydrogenase, myophosphorylase, phosphofructokinase, lactate dehydrogenase, and creatine kinase activities. Between-group analyses were adjusted for MHC-I %. No differences in aerobic capacity were found between groups. Despite subtle elevations of plasma glucose levels in the PPM+ group, there was no difference in MHC sub-types or muscle enzymatic activities between the malaria-exposed and non-exposed groups. The current study did not show differences in MHC towards glycolytic sub-types or enzymatic activity across the sub-groups. The results support the notion of the mild elevations of plasma glucose levels in people exposed to placental malaria in pregnancy being due to compromised pancreatic insulin secretion rather than insulin resistance.

Hematological, skeletal muscle fiber and exercise performance adaptations to heat training in elite female and male cyclists.

Heat exercise training may increase exercise performance in athletes. The underlying mechanisms remain partly unresolved, and it is unknown if female and male athletes may experience comparable gains. The aims were to investigate whether heat training (HEAT) increases hemoglobin mass (Hbmass), skeletal muscle fiber characteristics and thermoneutral exercise performance in elite female and male endurance athletes. Female (n=20; VO2max = 58.2 ± 6.7 ml.min.kg) and male (n=27; VO2max = 76.4 ± 7.8 ml.min.kg) cyclists were studied before and after five weeks of randomized control or HEAT training consisting of five weekly sessions each of 50 min duration which were included in their normal training regimes. Overall, the observed relative responses to HEAT were largely similar in female and male study participants. HEAT increased (P<0.05) Hbmass in female from 650 ± 77 to 675 ± 76 g (4.0 ± 1.6%) and from 1008 ± 155 to 1041 ± 147 g (3.5 ± 2.3%) in male. In contrast, skeletal muscle citrate synthase activity, fiber type distribution and capillary density remained unchanged with HEAT. Lactate threshold, VO2max and mean power output during 15 min all out testing were all enhanced (P<0.05) following HEAT in female and male study participants. In conclusion, five weeks of HEAT increases Hbmass in female and male elite cyclists and improves exercise performance in a thermoneutral environment. Based on this, heat training may be recommended to elite female and male athletes aiming to perform in a thermoneutral environment.

A genome-wide landscape of mRNAs, miRNAs, lncRNAs, and circRNAs of skeletal muscles during dietary restriction in Mongolian horses

The proportion of different muscle fibers is essential for the horse breed's aptitude for athletic activities. Adaptation of locomotor muscle is correlated with altered physiologic conditions. To investigate the adaptive changes of muscle fiber phenotype and transcriptome in horse skeletal muscle during dietary restriction (DR). The muscle fiber type distribution and deep RNA-seq analysis of detecting differentially expressed mRNAs (DEGs), miRNA (DEMIRs), lncRNAs (DELs), circRNAs (DECs), and their function analysis were investigated in gluteus medius muscle of Mongolian horses during DR. A total of 1433 DEGs, 5 DEMIRs, 329 DELs, and 53 DECs were identified. Differing from non-uniform muscle fiber type changing, functional enrichment analysis showed that most downregulated DEGs were associated in muscle contraction, fuel energy metabolism, and protein balance. Linkages between non-coding RNA and mRNA landscape were detected from their functional changes. Our study provides new insights into the expressional changes of mRNA and non-coding RNA in horse skeletal muscles during DR, which might improve our understanding of the molecular mechanisms regulating muscle adaption during DR for racing horses.

New tools for the investigation of muscle fiber-type spatial distributions across histological sections

BackgroundThe functional and metabolic properties of skeletal muscles are partly a function of the spatial arrangement of fibers across the muscle belly. Many muscles feature a non-uniform spatial pattern of fiber types, and alterations to the arrangement can reflect age or disease and correlate with changes in muscle mass and strength. Despite the significance of this event, descriptions of spatial fiber-type distributions across a muscle section are mainly provided qualitatively, by eye. Whilst several quantitative methods have been proposed, difficulties in implementation have meant that robust statistical analysis of fiber type distributions has not yielded new insight into the biological processes that drive the age- or disease-related changes in fiber type distributions.MethodsWe review currently available approaches for analysis of data reporting fast/slow fiber type distributions on muscle sections before proposing a new method based on a generalized additive model. We compare current approaches with our new method by analysis of sections of three mouse soleus muscles that exhibit visibly different spatial fiber patterns, and we also apply our model to a dataset representing the fiber type proportions and distributions of the mouse tibialis anterior.ResultsWe highlight how current methods can lead to differing interpretations when applied to the same dataset and demonstrate how our new method is the first to permit location-based estimation of fiber-type probabilities, in turn enabling useful graphical representation.ConclusionsWe present an open-access online application that implements current methods as well as our new method and which aids the interpretation of a variety of statistical tools for the spatial analysis of muscle fiber distributions.

MP03-06 VAGINAL DELIVERY ALTERS THE MUSCLE FIBER COMPOSITION OF THE PELVIC FLOOR MUSCLES

You have accessJournal of UrologyCME1 Apr 2023MP03-06 VAGINAL DELIVERY ALTERS THE MUSCLE FIBER COMPOSITION OF THE PELVIC FLOOR MUSCLES Takeya Kitta, Yui Abe-Takahashi, Mifuka Ouchi, Hiroki Chiba, Madoka Hoguchi, Mio Togo, Naohisa Kusakabe, Hidehiro Kakizaki, and Nobuo Shinohara Takeya KittaTakeya Kitta More articles by this author , Yui Abe-TakahashiYui Abe-Takahashi More articles by this author , Mifuka OuchiMifuka Ouchi More articles by this author , Hiroki ChibaHiroki Chiba More articles by this author , Madoka HoguchiMadoka Hoguchi More articles by this author , Mio TogoMio Togo More articles by this author , Naohisa KusakabeNaohisa Kusakabe More articles by this author , Hidehiro KakizakiHidehiro Kakizaki More articles by this author , and Nobuo ShinoharaNobuo Shinohara More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003214.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: It is well known that transvaginal childbirth weakens the pelvic floor muscles (PFM) and pelvic nerves, leading to stress urinary incontinence (SUI). PFM training is recommended as Grade A conservative therapy for SUI. Thus, PFM play important roles in lower urinary tract functions. In our previous study, we reported that the distribution of fiber types in the PFM is altered beginning 1 to 4 weeks post vaginal distension (VD), leading to muscle atrophy. SUI after vaginal delivery is known to persist in approximately 2% of patients who have had a vaginal delivery, even after 2 months postpartum. Therefore, we hypothesized that the long-term SUI that occurs after vaginal delivery may be influenced by the fiber-type distribution of PFM. The purpose of this study was to examine the long-term histological changes in PFM in a rat model of VD. METHODS: Eighteen female Sprague-Dawley rats (248-335 g) were used in this study. They were divided into 3 groups: a sham group, 1 (1M) and 3 months (3M) post VD. Urethral pressure was measured as urethral baseline pressure (UBP) and amplitude of urethral response (A-URE) (cmH2O). Right side pubococcygeus muscle (Pcm) was sampled and wet weight to body weight was measured. Pcm frozen sections 10 mm thick were prepared and stained using succinate dehydrogenase (SDH) activity. Stained images were imported into a personal computer to identify muscle fiber type (Ⅰ, Ⅱa and Ⅱb) distribution. In addition, the cross-sectional area (CSA) (μm2) of each muscle fiber type was determined. One-way analysis of variance was performed to compare the data among 3 groups. P value of less than 0.05 was regarded to be statistically significant. The present study was conducted after being approved by Animal Study Facility Ethics Committee. RESULTS: UBP was not significantly different among the 3 groups. In A-URE, 1M group was significantly lower than in the sham (p < 0.05). The Pcm wet weight was significantly lower in the 1M and 3M groups compared to the sham group (p<0.05). The CSA of type Ⅰ fibers in the Pcm was significantly lower in the 1M group than in the 3M group (p<0.05). The CSA of Type Ⅱa and Ⅱb fibers was not significantly different among the 3 groups. The ratio of typeⅠ fibers was significantly lower in the 1M and 3M groups compared to the sham group. The ratio of type Ⅱa fibers was not significantly different among the 3 groups. The ratio of type Ⅱb fibers was significantly higher in the 1M and 3M groups than in the sham group (p<0.05). CONCLUSIONS: In the present study, at 3 months post VD, the urethral function improved, but there was a decrease in muscle wet weight of the Pcm and a change in muscle fiber composition (leading to a decrease in slow-twitch muscle response). This may be due to decreased muscle function due to atrophy of the PFM and changes in muscle composition when SUI occurs beyond 3 months after vaginal delivery. Source of Funding: none © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e23 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Takeya Kitta More articles by this author Yui Abe-Takahashi More articles by this author Mifuka Ouchi More articles by this author Hiroki Chiba More articles by this author Madoka Hoguchi More articles by this author Mio Togo More articles by this author Naohisa Kusakabe More articles by this author Hidehiro Kakizaki More articles by this author Nobuo Shinohara More articles by this author Expand All Advertisement PDF downloadLoading ...

Normal muscle fiber type distribution is recapitulated in aged ephrin-A3-/- mice that previously lacked most slow myofibers.

Individual limb muscles have characteristic representation and spatial distribution of muscle fiber types (one slow and up to three fast isoforms) appropriate to their unique anatomical location and function. This distribution can be altered by physiological stimuli such as training (i.e., for increased endurance or force) or pathological conditions such as aging. Our group previously showed that ephrin-A3 is expressed only on slow myofibers, and that adult mice lacking ephrin-A3 have dramatically reduced numbers of slow myofibers due to postnatal innervation of previously slow myofibers by fast motor neurons. In this study, fiber type composition of hindlimb muscles of aged and denervated/reinnervated C57BL/6 and ephrin-A3-/- mice was analyzed to determine whether the loss of slow myofibers persists across the lifespan. Surprisingly, fiber-type composition of ephrin-A3-/- mouse muscles at two years of age was nearly indistinguishable from age-matched C57BL/6 mice. After challenge with nerve crush, the percentage of IIa and I/IIa hybrid myofibers increased significantly in aged ephrin-A3-/- mice. While EphA8, the receptor for ephrin-A3, is present at all neuromuscular junctions (NMJs) on fast fibers in 3-6 mo old C57BL/6 and ephrin-A3-/- mice, this exclusive localization is lost with aging, with EphA8 expression now found on a subset of NMJs on some slow muscle fibers. This return to appropriate fiber-type distribution given time and under use reinforces the role of activity in determining fiber-type representation and suggests that, rather than being a passive baseline, the developmentally and evolutionarily selected fiber type pattern may instead be actively reinforced by daily living.

Fiber-Type Shifting in Sarcopenia of Old Age: Proteomic Profiling of the Contractile Apparatus of Skeletal Muscles.

The progressive loss of skeletal muscle mass and concomitant reduction in contractile strength plays a central role in frailty syndrome. Age-related neuronal impairments are closely associated with sarcopenia in the elderly, which is characterized by severe muscular atrophy that can considerably lessen the overall quality of life at old age. Mass-spectrometry-based proteomic surveys of senescent human skeletal muscles, as well as animal models of sarcopenia, have decisively improved our understanding of the molecular and cellular consequences of muscular atrophy and associated fiber-type shifting during aging. This review outlines the mass spectrometric identification of proteome-wide changes in atrophying skeletal muscles, with a focus on contractile proteins as potential markers of changes in fiber-type distribution patterns. The observed trend of fast-to-slow transitions in individual human skeletal muscles during the aging process is most likely linked to a preferential susceptibility of fast-twitching muscle fibers to muscular atrophy. Studies with senescent animal models, including mostly aged rodent skeletal muscles, have confirmed fiber-type shifting. The proteomic analysis of fast versus slow isoforms of key contractile proteins, such as myosin heavy chains, myosin light chains, actins, troponins and tropomyosins, suggests them as suitable bioanalytical tools of fiber-type transitions during aging.

Metabolic dysregulation and decreased capillarization in skeletal muscles of male adolescent offspring rats exposed to gestational intermittent hypoxia.

Gestational intermittent hypoxia (IH) is a hallmark of obstructive sleep apnea that occurs frequently during pregnancy, and effects caused by this environmental change during pregnancy may be transmitted to the offspring. In this study, we aimed to clarify the effects of IH in pregnant rats on the skeletal muscle of adolescent offspring rats. Mother rats underwent IH from gestation day 7-21, and their 5-weeks-old male offspring were analyzed. All male offspring rats were born and raised under normoxia conditions. Although no general growth retardation was observed, we found that exposure to gestational IH reduces endurance running capacity of adolescent offspring rats. Both a respiratory muscle (diaphragm; DIA) and a limb muscle (tibialis anterior; TA) showed no histological abnormalities, including fiber size and fiber type distribution. To identify the possible mechanism underlying the reduced running capacity, regulatory factors associated with energy metabolism were analyzed in different parts of skeletal muscles. Compared with rats born under conditions of gestational normoxia, gestational IH offspring rats showed significantly lower expression of genes associated with glucose and lipid metabolism, and lower protein levels of phosphorylated AMPK and AKT. Furthermore, gene expression of adiponectin receptors one and two was significantly decreased in the DIA and TA muscles. In addition, the DIA muscle from adolescent rats had significantly decreased capillary density as a result of gestational IH. However, these changes were not observed in a sucking muscle (geniohyoid) and a masticating muscle (masseter) of these rats. These results suggest that respiratory and limb muscles are vulnerable to gestational IH, which induces altered energy metabolism with decreased aerobic motor function. These changes were partially owing to the decreased expression of adiponectin receptors and decreased capillary density in adolescent offspring rats.

In Adult Skeletal Muscles, the Co-Receptors of Canonical Wnt Signaling, Lrp5 and Lrp6, Determine the Distribution and Size of Fiber Types, and Structure and Function of Neuromuscular Junctions.

Canonical Wnt signaling is involved in skeletal muscle cell biology. The exact way in which this pathway exerts its contribution to myogenesis or neuromuscular junctions (NMJ) is a matter of debate. Next to the common co-receptors of canonical Wnt signaling, Lrp5 and Lrp6, the receptor tyrosine kinase MuSK was reported to bind at NMJs WNT glycoproteins by its extracellular cysteine-rich domain. Previously, we reported canonical Wnt signaling being active in fast muscle fiber types. Here, we used conditional Lrp5 or Lrp6 knockout mice to investigate the role of these receptors in muscle cells. Conditional double knockout mice died around E13 likely due to ectopic expression of the Cre recombinase. Phenotypes of single conditional knockout mice point to a very divergent role for the two receptors. First, muscle fiber type distribution and size were changed. Second, canonical Wnt signaling reporter mice suggested less signaling activity in the absence of Lrps. Third, expression of several myogenic marker genes was changed. Fourth, NMJs were of fragmented phenotype. Fifth, recordings revealed impaired neuromuscular transmission. In sum, our data show fundamental differences in absence of each of the Lrp co-receptors and suggest a differentiated view of canonical Wnt signaling pathway involvement in adult skeletal muscle cells.

Muscle fiber type-specific proteome distribution and protease activity in relation to proteolysis trends in beef striploin (M. longissimus lumborum) and tenderloin (M. psoas major)

The muscle fiber type and proteome distributions, protease activity, and protein degradation in bovine M. longissimus lumborum (beef striploin; LL) and M. psoas major (beef tenderloin; PM) were assessed to study muscle type-specific proteolysis. Bovine LL and PM muscle pieces (3 cm thickness) were stored in a refrigerator for 14 days and protein degradation and protease activity were evaluated at 1, 7, and 14 days of storage. Bovine LL included more fast-twitch glycolytic (type IIX) fibers than PM, whereas PM had a higher composition of slow-twitch oxidative (type I) fibers than LL (P < 0.05). Proteome distribution between these two muscles showed muscle fiber type-specificities: LL showed high intensities of proteins responsible for glycolytic metabolism, while PM showed large distribution of mitochondrial proteins. The activity of proteases, mainly calpain, showed different tendencies between the two muscles during 14 days of cold storage, which resulted in different timing of excessive myofibrillar protein breakdown (myosin heavy chains 1 and 2) between the two muscles. These results provided a detailed understanding of the differences in proteolysis by muscle type resulting in proteolysis-induced changes in meat quality during storage.

Association of Gene Variants for Mechanical and Metabolic Muscle Quality with Cardiorespiratory and Muscular Variables Related to Performance in Skiing Athletes.

Background: Skiing is a popular outdoor sport posing different requirements on musculoskeletal and cardiorespiratory function to excel in competition. The extent to which genotypic features contribute to the development of performance with years of ski-specific training remains to be elucidated. We therefore tested whether prominent polymorphisms in genes for angiotensin converting enzyme (ACE-I/D, rs1799752), tenascin-C (TNC, rs2104772), actinin-3 (ACTN3, rs1815739) and PTK2 (rs7460 and rs7843014) are associated with the differentiation of cellular hallmarks of muscle metabolism and contraction in high level skiers. Material & Methods: Forty-three skiers of a world-leading national ski team performed exhaustive cardiopulmonary exercise testing as well as isokinetic strength testing for single contractions, whereby 230 cardiopulmonary measurements were performed in the period from 2015–2018. A total of 168 and 62 data measurements were from the Alpine and Nordic skiing squads, respectively. Ninety-five and one hundred thirty-five measurements, respectively, were from male and female athletes. The average (±SD) age was 21.5 ± 3.0 years, height 174.0 ± 8.7 cm, and weight 71.0 ± 10.9 kg for the analysed skiers. Furthermore, all skiers were analysed concerning their genotype ACE-I/D, Tenascin C, ACTN3, PTK2. Results: The genotype distribution deviated from Hardy–Weinberg equilibrium for the ACTN3 genotype, where rs1815739-TT genotypes (corresponding to the nonsense mutation) were overrepresented in world-class skiers, indicating a slow muscle fibre phenotype. Furthermore, the heterozygous rs2104772-AT genotypes of TNC also demonstrated the best scaled peak power output values during ramp exercise to exhaustion. The highest values under maximum performance for heart rate were associated with the rs1799752-II and rs1815739-CC genotypes. The lowest values for peak power of single contractions were achieved for rs1815739-CC, rs1799752-II and rs7843014-CT genotypes. The skiing discipline demonstrated a main influence on cardiorespiratory parameters but did not further interact with genotype-associated variability in performance. Discussion: Classically, it is pointed out that muscles of, for example, alpine skiers do not possess a distinct fibre type composition, but that skiers tend to have a preponderance of slow-twitch fibres. Consequently, our findings of an overrepresentation of ACTN3-TT genotypes in a highly selective sample of elite world class skiers support the potential superiority of a slow fibre type distribution. Conclusions: We suggest that one competitive advantage that results from a slow, typically fatigue-resistant fibre type distribution might be that performance during intense training days is better preserved, whereby simply a higher technical training volume can be performed, yielding to a competitive advantage.

FP.16 The effects of resistance exercise training on mitochondrial myopathy patients

Patients with single, large-scale mitochondrial DNA (mtDNA) deletions often present with mitochondrial myopathy (MM) for which no treatment is available. Exercise intervention induces beneficial effects in mitochondrial myopathy. However, the mechanisms for this are not yet understood. The aim of this study is to investigate genetic, biochemical and molecular changes triggered by 16-week resistance exercise training for the development of exercise mimetic drugs for the cure of MM patients. Quadriceps biopsies were taken from a cohort of single, large-scale mtDNA deletion patients before and after 16-week resistance exercise training. Biopsies were assessed for changes after exercise in oxidative phosphorylation (OXPHOS) staining for Laminin, VDAC1, Ndufb8 and COX1; fibre type distribution, size and contraction force staining for Myh7, Myh2 and Myh1; heteroplasmy using real-time PCR for MT-ND4, MT-CO1, MT-CO3 and MT-CYB genes; differentially expressed (DE) genes and related protein level using RNA-sequencing and imaging mass cytometry, respectively. Myopathy score is a strong predictor of resistance exercise training responsiveness. Compared to healthy controls, all patients present with a significantly lower contraction force of SKM fibres (p=0.003) and differential fibre type distribution with increased type 2a (p<0.04) and decreased fibre I (p<0.0001). After exercise, all cases show an increase in the proportion of fibres with higher level of COX1 (p<0.0012) and increase of Myh7⁺ fibre type size (all patients except P2, p < 0.0001). The majority of patients show a significant increase of mitochondrial mass that positively correlates with the increase of aerobic capability (R=0.81). Furthermore, 13,132 DE genes were identified between pre- and post-exercise cases, of which 40 DE genes were selected with p<0.05 and logarithmic fold change <1.5 or >1.5. Among these, 13 novel exercise-modulated targets were established with an increased protein level localising within the SKM fibres (e.g. Dock11, Edil3, Ikbkg, Gem). Further analysis will assess the protein level of targets with nuclear, perinuclear and extracellular localisation in SKM of MM patients. Resistance exercise training is effective in reducing the OXPHOS defects and induces SKM remodelling in MM patients. The identification of novel exercise-modulated targets may provide insight for the development of exercise mimetic drugs for the cure of mitochondrial myopathy patients.

Proteolysis and changes in meat quality of chicken pectoralis major and iliotibialis muscles in relation to muscle fiber type distribution

The proteolysis trends and meat quality of the chicken pectoralis major (PM) and iliotibialis (IL) muscles stored at 4°C for 7 d were investigated. After 7 d of storage, the purge loss was higher (P < 0.05) in PM than in IL muscle. The difference in the composition of muscle fibers between PM (100% fast type) and IL (88.85% fast and 11.15% slow types) resulted in differences in proteolysis. Fructose-bisphosphate aldolase, troponin I, myosin heavy chain, and malate dehydrogenase exhibited the same tendencies, but pyruvate kinase, creatine kinase, L-lactate dehydrogenase, and triosephosphate isomerase exhibited different tendencies in the 2 muscles. The activity of cathepsin B was higher in PM than in IL during storage (P < 0.05). These results indicate that the proteolysis trend and changes in meat quality during cold storage are dependent on the different muscle fiber characteristics.

Role of progression of training volume on intramuscular adaptations in patients with chronic obstructive pulmonary disease.

Introduction: Quadriceps dysfunction is a common systemic manifestation of chronic obstructive pulmonary disease (COPD), for which treatment using resistance training is highly recommended. Even though training volume is suggested to be a key explanatory factor for intramuscular adaptation to resistance training in healthy older adults, knowledge is scarce on the role of progression of training volume for intramuscular adaptations in COPD. Methods: This study was a sub-analysis of a parallel-group randomized controlled trial. Thirteen patients with severe to very severe COPD (median 66 yrs, forced expiratory volume in 1 s 44% predicted) performed 8 weeks of low-load resistance training. In a post hoc analysis, they were divided into two groups according to their training volume progression. Those in whom training volume continued to increase after the first 4 weeks of training outlined the continued progression group (n = 9), while those with limited increase (<5%) or even reduction in training volume after the initial 4 weeks composed the discontinued progression group (n = 4). Fiber-type distribution and oxidative muscle protein levels, i.e., citrate synthase (CS), hydroxyacyl-coenzyme A dehydrogenase (HADH), mitochondrial transcription factor A (TfAM) as well as quadriceps endurance measures (total work from elastic band and isokinetic knee extension tests), were assessed before and after the intervention period. Results: The continued progression group sustained their training volume progression during weeks 5–8 compared to weeks 1–4 (median +25%), while the discontinued progression group did not (median -2%) (p = 0.007 between groups). Compared with baseline values, significant between-group differences in fiber type distribution and TfAM muscle protein levels (range ± 17–62%, p < 0.05) and in individual responses to change in Type I and Type IIa fiber type proportion, CS, HADH, and TfAM muscle protein levels outcomes (median 89 vs. 50%, p = 0.001) were seen in favor of the continued progression group. Moreover, only the continued progression group had a significant increase in HADH muscle protein levels (+24%, p = 0.004), elastic band (+56%, p = 0.004) and isokinetic (+7%, p = 0.004) quadriceps endurance, but the between-group differences did not reach statistical significance (range 14–29%, p = 0.330–1.000). Discussion: The novel findings of the current study were that patients with COPD who had a continued progression of training volume across the 8-weeks intervention had an increased proportion of Type I fibers, and TfAM muscle protein levels and decreased proportion of Type II fibers compared to those that did not continue to progress their training volume after the initial weeks. Additionally, HADH muscle protein levels and quadriceps endurance measurements only improved in the continued progression group, although no significant between-group differences were seen. These findings highlight the importance of continued progression of training volume during resistive training to counteract quadriceps dysfunction within the COPD population. Still, considering the small sample size and the post hoc nature of our analyses, these results should be interpreted cautiously, and further research is necessary.