Myosin Heavy Chain Composition Research Articles

Kbtbd13R408C-knockin mouse model reveals impaired relaxation kinetics as novel pathomechanism for NEM6 cardiomyopathy

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): VENI-Nederlandse organisatie voor Wetenschappelijk Onderzoek (NWO) We identified a novel cardiomyopathy gene: KBTBD13 encoding kelch repeat and BTB (POZ) domain containing 13. The Dutch founder variant KBTBD13R408C not only affects skeletal muscle function resulting in nemaline myopathy type 6 (NEM6), but also affects the heart. In the Dutch NEM6 cohort, various cardiac abnormalities were observed, including systolic dysfunction, diastolic dysfunction, atrial fibrillation, ventricular tachycardia and three patients died of sudden cardiac death. The majority of NEM6 patients harbors the Dutch founder variant, c.1222C>T, p.Arg408Cys (KBTBD13R408C). To provide insight in the mechanism underlying cardiac dysfunction in NEM6, here we assessed cardiac structure and function in Kbtbd13R408C-knockin mice, which closely recapitulate the well characterized skeletal muscle phenotype, i.e. impaired relaxation kinetics and mitochondrial dysfunction. Pressure-volume loop analyses showed that the end-diastolic pressure-volume relation was steeper in Kbtbd13R408C-knockin mice, indicating diastolic dysfunction. Histological evaluation of cardiac structure revealed no increase in fibrosis in Kbtbd13R408C-knockin mice. Also, no alterations in titin isoform composition or myosin heavy chain composition were observed that could account for the increased diastolic stiffness. Next, we studied the contraction and relaxation kinetics in wild type (WT) and Kbtbd13R408C-knockin mice at the intact single cardiomyocyte level by a high-throughput contractility set-up. Our data showed a prolonged relaxation time in Kbtbd13R408C-knockin mice compared to WT mice, indicating impaired relaxation kinetics. No differences in the magnitude of contraction (percentage of shortening) was found between cardiomyocytes of Kbtbd13R408C-knockin and WT mice. In parallel, calcium-handling was assessed by calcium indicator Fura-2AM. These studies reveal that calcium-release is not affected, but calcium-reuptake is slower in Kbtbd13R408C-knockin mice, which might contribute to the impaired relaxation kinetics. Current studies focus on how KBTBD13R408C affects calcium-reuptake kinetics and sarcomere kinetics in NEM6 cardiomyocytes. Hence, our studies provide the first insights in the pathomechanism underlying cardiac dysfunction in NEM6.

Physiological and molecular predictors of cycling sprint performance.

The study aimed to identify novel muscle phenotypic factors that could determine sprint performance using linear regression models including the lean mass of the lower extremities (LLM), myosin heavy chain composition (MHC), and proteins and enzymes implicated in glycolytic and aerobic energy generation (citrate synthase, OXPHOS proteins), oxygen transport and diffusion (myoglobin), ROS sensing (Nrf2/Keap1), antioxidant enzymes, and proteins implicated in calcium handling. For this purpose, body composition (dual-energy X-ray absorptiometry) and sprint performance (isokinetic 30-s Wingate test: peak and mean power output, Wpeak and Wmean ) were measured in young physically active adults (51 males and 10 females), from which a resting muscle biopsy was obtained from the musculus vastus lateralis. Although females had a higher percentage of MHC I, SERCA2, pSer16 /Thr17 -phospholamban, and Calsequestrin 2 protein expressions (all p < 0.05), and 18.4% lower phosphofructokinase 1 protein expression than males (p < 0.05), both sexes had similar sprint performance when it was normalized to body weight or LLM. Multiple regression analysis showed that Wpeak could be predicted from LLM, SDHB, Keap1, and MHC II % (R 2 = 0.62, p < 0.001), each variable contributing to explain 46.4%, 6.3%, 4.4%, and 4.3% of the variance in Wpeak , respectively. LLM and MHC II % explained 67.5% and 2.1% of the variance in Wmean , respectively (R 2 = 0.70, p < 0.001). The present investigation shows that SDHB and Keap1, in addition to MHC II %, are relevant determinants of peak power output during sprinting.

Age-related sex differences in tongue strength and muscle morphometry in a rat model

ObjectiveTo investigate potential effects of sex on voluntary tongue strength, evoked twitch and tetanic tension, speed of contraction, and muscle fiber cross-sectional area in the muscles of the rat tongue. Additionally, we aimed to determine whether estrous cycle stage impacts any of the dependent variables as a pilot investigation into the use of female rats in a model of tongue exercise and aging. DesignFischer 344-Brown Norway male and female rats in two age groups (16 middle-aged, 16 young-adult) were trained to use a tongue force operandum. Tongue muscle contraction, myosin heavy chain (MyHC) composition, and cross section area of the genioglossus and styloglossus muscles were examined. Vaginal lavage determined estrous cycle stage of the female rats daily. ResultsThe female group had significantly lower evoked twitch and tetanic tension, longer contraction times, and a smaller proportion of MyHC type IIa and MyHC type IIx in the styloglossus muscle. There was no significant sex effect in maximal voluntary tongue force (MVTF) despite a significant weight difference between the male and female groups. There were no significant age or sex effects in the genioglossus. Estrous cycle stage did not have a significant effect on any of the dependent variables. ConclusionsSex and age both have a significant effect on tongue muscle structure and physiology. While the female group showed reduced contraction speed and maximal twitch and tetanic tension relative to the male group, differences in muscle morphology appeared to vary by muscle.

Histological and contractile changes in the genioglossus muscle after nasal obstruction in growing rats

The aim of the study was to address the genioglossus muscle physiological and histological changes after unilateral nasal obstruction in growing rats. Fifty-four 6-day-old male Wistar albino rats were randomly divided into control (n = 27) and experimental (n = 27) groups. Unilateral nasal obstruction was performed at 8 days old. Contractile properties of the genioglossus whole muscle were measured at 5-, 7- and 9-week-old, including the twitch and tetanic forces, contraction time, half-decay time, and fatigue index. The histological characteristics of the genioglossus were also evaluated at 5-, 7- and 9-week-old, analyzing the myosin heavy chain composition of the slow, fast, IIa and IIb muscle fiber type, by measuring the number, rate, diameter and cross-sectional area. The maximal twitch force, and tetanic force at 60 Hz and 80 Hz force was significantly increased at all ages after nasal obstruction. The fatigue index was decreased at 5 weeks-old after nasal obstruction. The diameter and cross-sectional area of the fast, IIa and IIb muscle fiber types were increased at 7 and 9 weeks after nasal obstruction, while only the diameter of IIa type and cross-sectional area of IIb type were increased at 5 weeks-old after nasal obstruction. Nasal obstruction during growth affects the whole genioglossus muscle contractile properties and histological characteristics, increasing its force, the diameter and area of its muscle fibers. These changes in the genioglossus muscle may affect the normal growth, development and function of the craniofacial complex.

Type IIa Muscle Fibers Influence The Force-velocity Profile Of Healthy Adults During Maximal Cycling

During maximal cycling, vastii muscles contract during the extension phase of the pedaling cycle to generate a large portion of pedal forces. The contractile properties of the vastii muscles vary between individuals, depending on myosin heavy chain (MHC) composition. A positive association was reported between the cross-sectional area of type II muscle fibers in vastus lateralis (VL) and optimal cadence (Copt) during maximal cycling derived from power-cadence (P-C) relationships computed over the full pedal cycle. However, the contribution of vastii muscles to power produced over the full cycle may also vary across individuals due to potential differences in the relative contribution of the flexor and extensor muscles of the left and right lower limbs. PURPOSE: To investigate if MHC composition in VL influences variables derived from P-C relationships computed over the extension phase during maximal cycling. METHODS: Thirteen healthy adults (9 males, 4 females) performed a Force-Velocity test on a stationary cycle ergometer. We modelled individual P-C relationships using third order polynomial regressions to extract maximal power (Pmax), Copt and maximal cadence (C0) for the extension phase of the right leg only. From a muscle biopsy of the right vastus lateralis, we used electrophoresis to determine the MHC isoform composition (I, IIa and IIx). Pearson correlations were determined between % of each MHC type (I, IIa, IIx) and the variables derived from the P-C relationships (Pmax, Copt and C0). RESULTS: All individual P-C relationships were well fitted (r2 = 0.92 ± 0.01), allowing extraction of Pmax (11.9 ± 2.6 W/kg), Copt (130 ± 11 rpm) and C0 (235 ± 25 rpm). MHC isoform composition comprised 44 ± 10% type I, 47 ± 12% type IIa and 9 ± 5% IIx. Type I % MHC was negatively associated with Pmax (r = -0.553, p = 0.05) and Copt (r = -0.737, p < 0.05). Type IIa % MHC was positively associated with Pmax (r = 0.623, p < 0.05), Copt (r = 0.849, p < 0.001) and C0 (r = 0.805, p < 0.01). No significant associations were found between type IIx % MHC and Pmax, Copt or C0 (all P > 0.05). CONCLUSIONS: Healthy adults with higher type IIa % MHC in the VL muscle produced greater power at higher optimal cadences and reached higher maximal cadences during maximal cycling, when P-C relationships were computed over the extension phase of the ipsilateral limb.

Skeletal muscle undergoes fiber type metabolic switch without myosin heavy chain switch in response to defective fatty acid oxidation

ObjectiveSkeletal muscle is a heterogeneous and dynamic tissue that adapts to functional demands and substrate availability by modulating muscle fiber size and type. The concept of muscle fiber type relates to its contractile (slow or fast) and metabolic (glycolytic or oxidative) properties. Here, we tested whether disruptions in muscle oxidative catabolism are sufficient to prompt parallel adaptations in energetics and contractile protein composition. MethodsMice with defective mitochondrial long-chain fatty acid oxidation (mLCFAO) in the skeletal muscle due to loss of carnitine palmitoyltransferase 2 (Cpt2Sk−/−) were used to model a shift in muscle macronutrient catabolism. Glycolytic and oxidative muscles of Cpt2Sk−/− mice and control littermates were compared for the expression of energy metabolism-related proteins, mitochondrial respiratory capacity, and myosin heavy chain isoform composition. ResultsDifferences in bioenergetics and macronutrient utilization in response to energy demands between control muscles were intrinsic to the mitochondria, allowing for a clear distinction of muscle types. Loss of CPT2 ablated mLCFAO and resulted in mitochondrial biogenesis occurring most predominantly in oxidative muscle fibers. The metabolism-related proteomic signature of Cpt2Sk−/− oxidative muscle more closely resembled that of glycolytic muscle than of control oxidative muscle. Respectively, intrinsic substrate-supported mitochondrial respiration of CPT2 deficient oxidative muscles shifted to closely match that of glycolytic muscles. Despite this shift in mitochondrial metabolism, CPT2 deletion did not result in contractile-based fiber type switching according to myosin heavy chain composition analysis. ConclusionThe loss of mitochondrial long-chain fatty acid oxidation elicits an adaptive response involving conversion of oxidative muscle toward a metabolic profile that resembles a glycolytic muscle, but this is not accompanied by changes in myosin heavy chain isoforms. These data suggest that shifts in muscle catabolism are not sufficient to drive shifts in the contractile apparatus but are sufficient to drive adaptive changes in metabolic properties.

Morphometric and Immunohistochemical Characteristics of the Adult Human Soft Palate Muscles.

The soft palate is the only structure that reversibly separates the respiratory and gastrointestinal systems. Most species can eat and breathe at the same time. Humans cannot do this and malfunction of the soft palate may allow food to enter the lungs and cause fatal aspiration pneumonia. Speech is the most defining characteristic of humans and the soft palate, along with the larynx and tongue, plays the key roles. In addition, palatal muscles are involved in snoring and obstructive sleep apnea. Considering the significance of the soft palate, its function is insufficiently understood. The objectives of this study were to document morphometric and immunohistochemical characteristics of adult human soft palate muscles, including fiber size, the fiber type, and myosin heavy chain (MyHC) composition for better understanding muscle functions. In this study, 15 soft palates were obtained from human autopsies. The palatal muscles were separated, cryosectioned, and stained using histological and immunohistochemical techniques. The results showed that there was a fast type II predominance in the musculus uvulae and palatopharyngeus and a slow type I predominance in the levator veli palatine. Approximately equal proportions of type I and type II fibers existed in both the palatoglossus and tensor veli palatine. Soft palate muscles also contained hybrid fibers and some specialized myofibers expressing slow-tonic and embryonic MyHC isoforms. These findings would help better understand muscle functions.

Do skeletal muscle composition and gene expression as well as acute exercise-induced serum adaptations in older adults depend on fitness status?

BackgroundInactive physical behavior among the elderly is one risk factor for cardiovascular disease, immobility and increased all-cause mortality. We aimed to answer the question whether or not circulating and skeletal muscle biomarkers are differentially expressed depending on fitness status in a group of elderly individuals.MethodsTwenty-eight elderly individuals (73.36 ± 5.46 years) participated in this exploratory study after participating as part of the multinational SITLESS-clinical trial (implementation of self-management and exercise programs over 16 weeks). A cardiopulmonary exercise test (CPX) and resting skeletal muscle biopsy were performed to determine individual physiological performance capacity. Participants were categorized into a high physical fitness group (HPF) and a low physical fitness group (LPF) depending on peak oxygen uptake (VO2peak). Serum blood samples were taken before (pre) and after (post) CPX and were examined regarding serum BDNF, HSP70, Kynurenine, Irisin and Il-6 concentrations. Skeletal muscle tissue was analyzed by silver staining to determine the myosin heavy chain (MyHC) composition and selected genes by qRT-PCR.ResultsHPF showed lower body weight and body fat, while skeletal muscle mass and oxygen uptake at the first ventilatory threshold (VO2T1) did not differ between groups. There were positive associations between VO2peak and VO2VT1 in HPF and LPF. MyHC isoform quantification revealed no differences between groups. qRT-PCR showed higher expression of BDNF and BRCA1 in LPF skeletal muscle while there were no differences in other examined genes regarding energy metabolism. Basal serum concentrations of Irisin were higher in HPF compared to LPF with a trend towards higher values in BDNF and HSP70 in HPF. Increases in Il-6 in both groups were observed post.ConclusionsAlthough no association between muscle composition/VO2peak with fitness status in older people was detected, higher basal Irisin serum levels in HPF revealed slightly beneficial molecular serum and muscle adaptations.Trial registrationClinicalTrials.gov, NCT02629666. Registered 19 November 2015.

Effects of Cardiac Myosin Heavy Chain Isoform Composition on Contraction Kinetics and Calcium Transients in Adult Rat Cardiomyocytes

Myosin is an essential structural and functional protein of the sarcomere. In rat ventricular myocardium two isoforms of myosin heavy chain (MyHC) are expressed, α-MyHC and β-MyHC. The two MyHC isoforms show different cross bridge kinetics. Strikingly, in the ventricular rat myocardium, cardiomyocytes with different MyHC composition are connected and contract without producing functional disbalances.

Ractopamine-induced fiber type-specific gene expression in porcine skeletal muscles is independent of growth.

Feeding ractopamine (RAC), a β-adrenergic agonist (BAA), to pigs increases type IIB muscle fiber type-specific protein and mRNA expression. However, increases in the abundance of these fast-twitch fiber types occur with other forms of muscle hypertrophy and thus BAA-induced changes in myosin heavy chain (MyHC) composition may simply be associated with increased muscle growth known to occur in response to BAA feeding. The objective of this study was to determine whether RAC feeding could change the MyHC gene expression in the absence of maximal muscle growth. Pigs were fed either an adequate diet that supported maximal muscle hypertrophy or a low nutrient diet that limited muscle growth. RAC was included in diets at 0 or 20 mg/kg for 1, 2, or 4 wk. Backfat depth was less (P < 0.05) in pigs fed the low nutrient diet compared with the adequate diet but was not affected by RAC. Loin eye area was greater (P < 0.05) in pigs fed an adequate diet plus RAC at 1 wk but did not differ among remaining pigs. At 2 and 4 wk, however, pigs fed the adequate diet had greater loin eye areas (P < 0.05) than pigs fed the low nutrient diet regardless of RAC feeding. Gene expression of the MyHC isoforms, I, IIA, IIX, and IIB, as well as glycogen synthase, citrate synthase, β 1-adrenergic receptor (AR), and β 2-AR were determined in longissimus dorsi (LD) and red (RST) and white (WST) portions of the semitendinosus muscles. MyHC type I gene expression was not altered by RAC or diet. Feeding RAC decreased (P < 0.01) MyHC type IIA gene expression in all muscles, but to a greater extent in WST and LD. MyHC type IIX gene expression was lower (P < 0.05) in WST and LD muscles in response to RAC but was not altered in RST muscles. RAC increased (P < 0.05) MyHC type IIB gene expression in all muscles, but to a greater extent in RST. β 1-AR gene expression was unaffected by RAC or diet, whereas the expression of the β 2-AR gene was decreased (P < 0.001) by RAC. No significant RAC * diet interactions were observed in gene expression in this study, indicating that RAC altered MyHC and β 2-AR gene expression in porcine skeletal muscles independent of growth.

Ketogenic diet feeding improves aerobic metabolism property in extensor digitorum longus muscle of sedentary male rats.

Recent studies of the ketogenic diet, an extremely high-fat diet with extremely low carbohydrates, suggest that it changes the energy metabolism properties of skeletal muscle. However, ketogenic diet effects on muscle metabolic characteristics are diverse and sometimes countervailing. Furthermore, ketogenic diet effects on skeletal muscle performance are unknown. After male Wistar rats (8 weeks of age) were assigned randomly to a control group (CON) and a ketogenic diet group (KD), they were fed for 4 weeks respectively with a control diet (10% fat, 10% protein, 80% carbohydrate) and a ketogenic diet (90% fat, 10% protein, 0% carbohydrate). After the 4-week feeding period, the extensor digitorum longus (EDL) muscle was evaluated ex vivo for twitch force, tetanic force, and fatigue. We also analyzed the myosin heavy chain composition, protein expression of metabolic enzymes and regulatory factors, and citrate synthase activity. No significant difference was found between CON and KD in twitch or tetanic forces or muscle fatigue. However, the KD citrate synthase activity and the protein expression of Sema3A, citrate synthase, succinate dehydrogenase, cytochrome c oxidase subunit 4, and 3-hydroxyacyl-CoA dehydrogenase were significantly higher than those of CON. Moreover, a myosin heavy chain shift occurred from type IIb to IIx in KD. These results demonstrated that the 4-week ketogenic diet improves skeletal muscle aerobic capacity without obstructing muscle contractile function in sedentary male rats and suggest involvement of Sema3A in the myosin heavy chain shift of EDL muscle.

Myosin heavy chain expression relationships to power-load and velocity-load curves

Velocity- and power-based training are popular methods of determining training session loads and volumes. One factor that may influence load-velocity and load-power properties of an individual is the myosin heavy chain (MHC) composition of the muscle. The aim of this study was to examine the relationship between MHC composition and both load-velocity and load-power properties of muscle performance. Forty-two men with a variety of training backgrounds took part in this study (mean±SD; age=22.4±3.5 yrs, hgt=1.78±0.07 m, BW=78.7±13.3 kg). After testing leg extension one repetition maximum (1 RM), subjects performed maximal effort leg extensions at loads from 30% to 90% 1 RM. Muscle biopsies from the vastus lateralis were analyzed via SDS-PAGE electrophoresis technique for MHC content (IIx=13.8±12.9%, IIa=49.4±10.3%, I=36.8±11.3%). Leg extension rotational velocity and power were plotted against relative loads for all subjects. Significant correlations (P<0.05) were observed for MHC IIa with all performance variables (i.e. slopes, intercepts, peaks and relative loads). Relationships indicated that greater %MHC IIa was associated with greater velocity intercepts, more negative load-velocity slopes, greater maximal power, and with maximal power occurring at a lower relative intensity (% 1 RM). These data indicate that muscle velocity and power characteristics appear to be partially influenced by MHC content in a manner consistent with single muscle fiber contractile properties.

Role of CaMKII and sarcolipin in muscle adaptations to strength training with different levels of fatigue in the set.

Strength training promotes a IIX-to-IIA shift in myosin heavy chain (MHC) composition, likely due to changes in sarcoplasmic [Ca2+ ] which are sensed by CaMKII. Sarcoplasmic [Ca2+ ] is in part regulated by sarcolipin (SLN), a small protein that when overexpressed in rodents stimulates mitochondrial biogenesis and a fast-to-slow fiber type shift. The purpose of this study was to determine whether CaMKII and SLN are involved in muscle phenotype and performance changes elicited by strength training. Twenty-two men followed an 8-week velocity-based resistance training program using the full squat exercise while monitoring repetition velocity. Subjects were randomly assigned to two resistance training programs differing in the repetition velocity loss allowed in each set: 20% (VL20) vs 40% (VL40). Strength training caused muscle hypertrophy, improved 1RM and increased total CaMKII protein expression, particularly of the δD isoform. Phospho-Thr287 -CaMKII δD expression increased only in VL40 (+89%), which experienced greater muscle hypertrophy, and a reduction in MHC-IIX percentage. SLN expression was increased in VL20 (+33%) remaining unaltered in VL40. The changes in phospho-Thr287 -CaMKII δD were positively associated with muscle hypertrophy and the number of repetitions during training, and negatively with the changes in MHC-IIX and SLN. Most OXPHOS proteins remained unchanged, except for NDUFB8 (Complex I), which was reduced after training (-22%) in both groups. The amount of fatigue allowed in each set critically influences muscle CaMKII and SLN responses and determines muscle phenotype changes. With lower intra-set fatigue, the IIX-to-IIA MHC shift is attenuated.

Myosin Heavy Chain Composition, Creatine Analogues, and the Relationship of Muscle Creatine Content and Fast-Twitch Proportion to Wilks Coefficient in Powerlifters.

Machek, SB, Hwang, PS, Cardaci, TD, Wilburn, DT, Bagley, JR, Blake, DT, Galpin, AJ, and Willoughby, DS. Myosin heavy chain composition, creatine analogues, and the relationship of muscle creatine content and fast-twitch proportion to Wilks coefficient in powerlifters. J Strength Cond Res 34(11): 3022-3030, 2020-Little data exist on powerlifting-specific skeletal muscle adaptations, and none elucidate sex differences in powerlifters. Powerlifters tend to display higher fast-twitch fiber content and phosphagen system dependence. Nevertheless, it is unknown whether fast-twitch fiber or muscle creatine content are predictive of competitive powerlifting performance (via Wilks coefficient). Twelve actively competing powerlifters (PL; n = 6M/6F; age = 21.3 ± 1.0; 3.0 ± 1.8 year competing; 7.3 ± 6.6 meets attended) and 10 sedentary controls (CON; n = 5M/5F; age = 19.4 ± 2.0 year) underwent vastus lateralis muscle biopsies and venipuncture to compare the myosin heavy chain (MHC) fiber type and creatine analogue profiles between groups of both sexes, and determine whether MHC IIa and muscle total creatine (MTC) composition predict powerlifting performance. Samples were analyzed for specific MHC isoform (I, IIa, and IIx) content via mixed homogenate SDS-PAGE, and creatine analogues (MTC, muscle creatine transporter [SLC6A8], serum total creatine [STC], and serum creatinine [CRT]). Furthermore, MHC IIa and MTC content were compared with Wilks coefficient using Pearson correlation coefficients. Male PL MHC content was 50 ± 6% I, 45 ± 6% IIa, and 5 ± 11% IIx, versus 46 ± 6% I, 53 ± 6 IIa, and 0% IIx in female PL. Conversely, male CON MHC content was 33 ± 5% I, 38 ± 7% IIa, and 30 ± 8% IIx, vs. 35 ± 9% I, 44 ± 8% IIa, and 21 ± 17% IIx in female CON. Muscle total creatine, SLC6A8, STC, and CRT did not significantly differ between groups nor sexes. Finally, neither MHC IIa content (r = -0.288; p = 0.364) nor MTC (r = 0.488; p = 0.108) significantly predicted Wilks coefficient, suggesting these characteristics alone do not determine powerlifting skill variation.

Myosin heavy chains in extraocular muscle fibres: Distribution, regulation and function.

This review examines kinetic properties and distribution of the 11 isoforms of myosin heavy chain (MyHC) expressed in extraocular muscle (EOM) fibre types and the regulation and function of these MyHCs. Although recruitment and discharge characteristics of ocular motoneurons during fixation and eye movements are well documented, work directly linking these properties with motor unit contractile speed and MyHC composition is lacking. Recruitment of motor units according to Henneman's size principle has some support in EOMs but needs consolidation. Both neurogenic and myogenic mechanisms regulate MyHC expression as in other muscle allotypes. Developmentally, multiply-innervated (MIFs) and singly-innervated fibres (SIFs) are derived presumably from distinct myoblast lineages, ending up expressing MyHCs in the slow and fast ends of the kinetic spectrum respectively. They modulate the synaptic inputs of their motoneurons through different retrogradely transported neurotrophins, thereby specifying their tonic and phasic impulse patterns. Immunohistochemical analyses of EOMs regenerating in situ and in limb muscle beds suggest that the very impulse patterns driving various ocular movements equip effectors with appropriate MyHC compositions and speeds to accomplish their tasks. These experiments also suggest that satellite cells of SIFs and MIFs are distinct lineages expressing different MyHCs during regeneration. MyHC compositions and functional characteristics of orbital fibres show longitudinal variations that facilitate linear ocular rotation during saccades. Palisade endings on global MIFs are postulated to respond to active and passive tensions by triggering axon reflexes that play important roles during fixation, saccades and vergence. How EOMs implement Listings law during ocular rotation is discussed.

Myosin heavy chain isoform composition in the deltoid and vastus lateralis muscles of elite handball players

ABSTRACT The purpose of the present study was to compare the myosin heavy chain (MHC) isoform composition of the deltoid and vastus lateralis muscles of the dominant and non-dominant limbs in handball players. Eleven male Greek elite handball players (age 22.6 ± 1.9 yrs, training experience 10.6 ± 2.1 yrs, height 184.1 ± 4.1 cm, and weight 81.0 ± 12.5 kg) participated in the study. Four muscle biopsies were obtained from the dominant and non-dominant deltoid and vastus lateralis muscles during the in-season period. The MHC composition was determined using SDS-PAGE. No significant difference was found between the dominant and non-dominant muscles; Deltoid muscle: MHC I [(95%CI = −1.22, 0.33), P = 0.228], MHC ΙΙa [(95%CI = −0.32, 1.59), P = 0.168] and MHC IIx [(95%CI = −1.49, 1.10), P = 0.749]; Vastus lateralis muscle: MHC I [(95%CI = −0.38, 0.63), P = 0.586], MHC ΙΙa [(95%CI = −0.50, 0.65), P = 0.783] and MHC IIx [(95%CI = −1.08, 0.42), P = 0.355]. The findings of the present study indicate that the greater use of the dominant limbs for throwing actions and body movements in handball do not lead to altered MHC isoform composition compared to the non-dominant limbs.

Four‐week Ketogenic Diet Improves Aerobic Metabolic Capacity in Extensor Digitrum Longus Muscle of Sedentary Male Rats

Ketogenic diet is an extremely high‐fat diet with low or absence of carbohydrate in the total nutrient. Thus, this diet causes ketone body production from the liver and shifts energy sources from carbohydrate to fat. Whereas previous studies have suggested that ketogenic diet ameliorates brain disorder such as drug‐resistant epilepsy, its influence on peripheral tissue is unclear. Skeletal muscle is an important organ for locomotion and metabolism. Recent studies have suggested that the ketogenic diet changes the energy metabolic properties in the skeletal muscle as well; however, the results are contradictory. Moreover, it remains unknown whether the ketogenic diet affects the muscle contractile function. Therefore, this study aimed to clarify the effect of ketogenic diet on muscle performance and biochemical adaptation in rat skeletal muscle. This study was approved by Institute for Animal Experimentation at St. Marianna University School of Medicine. Male Wistar rats (8 weeks of age) were assigned to either control group (CON) or ketogenic diet group (KD). CON and KD were fed for 4 weeks either control diet (10% fat, 10% protein, 80% carbohydrate) or ketogenic diet (90% fat, 10% protein, 0% carbohydrate), respectively. Body mass was recorded once a week. After the 4‐week feeding period, the animals were anesthetized, and the blood sample was collected to analyze blood ketone concentration. The right extensor digitrum longus (EDL) muscle was used to evaluate the twitch force, tetanic force, and fatigue ex vivo. The left EDL was homogenized and used to analyze the myosin heavy chain composition, the protein expression of metabolic enzymes, and the citrate synthase activity. Body mass was lower in KD than CON at third and fourth week of the feeding period. Blood ketone concentration in KD was significantly higher than CON. There was no statistical difference in the twitch and tetanic forces as well as muscle fatigue between CON and KD. However, the citrate synthase activity in KD was significantly higher than CON. The protein expression of citrate synthase, succinate dehydrogenase, cytochrome c oxidase subunit 4, and 3‐hydroxyacyl‐CoA dehydrogenase in KD was significantly higher than CON. Moreover, there was a shift of myosin heavy chain from type IIb to IIx in KD. Therefore, we conclude that 4‐week ketogenic diet improves skeletal muscle aerobic capacity without obstructing muscle contractile function in the sedentary male rats.Support or Funding InformationThis work was supported by MEXT/JSPS KAKENHI Grant Numbers JP24650334, JP18K17767.

In vivo characterization of skeletal muscle function in nebulin-deficient mice.

The conditional nebulin knockout mouse is a new model mimicking nemaline myopathy, a rare disease characterized by muscle weakness and rods within muscle fibers. We investigated the impact of nebulin (NEB) deficiency on muscle function in vivo. Conditional nebulin knockout mice and control littermates were studied at 10 to 12 months. Muscle function (force and fatigue) and anatomy (muscles volume and fat content) were measured in vivo. Myosin heavy chain (MHC) composition and nebulin (NEB) protein expression were assessed by protein electrophoresis. Conditional nebulin knockout mice displayed a lower NEB level (-90%) leading to a 40% and 45% reduction in specific maximal force production and muscles volume, respectively. Nebulin deficiency was also associated with higher resistance to fatigue and increased MHC I content. Adult nebulin-deficient mice displayed severe muscle atrophy and weakness in vivo related to a low NEB content but an improved fatigue resistance due to a slower contractile phenotype.

Medial gastrocnemius muscles fatigue but do not atrophy in paralyzed cat hindlimb after long-term spinal cord hemisection and unilateral deafferentation

This study of medial gastrocnemius (MG) muscle and motor units (MUs) after spinal cord hemisection and deafferentation (HSDA) in adult cats, asked 1) whether the absence of muscle atrophy and unaltered contractile speed demonstrated previously in HSDA-paralyzed peroneus longus (PerL) muscles, was apparent in the unloaded HSDA-paralyzed MG muscle, and 2) how ankle unloading impacts MG muscle and MUs after dorsal root sparing (HSDA-SP) with foot placement during standing and locomotion. Chronic isometric contractile forces and speeds were maintained for up to 12 months in all conditions, but fatigability increased exponentially. MU recordings at 8–11½ months corroborated the unchanged muscle force and speed with significantly increased fatigability; normal weights of MG muscle confirmed the lack of disuse atrophy. Fast MUs transitioned from fatigue resistant and intermediate to fatigable accompanied by corresponding fiber type conversion to fast oxidative (FOG) and fast glycolytic (FG) accompanied by increased GAPDH enzyme activity in absolute terms and relative to oxidative citrate synthase enzyme activity. Myosin heavy chain composition, however, was unaffected. MG muscle behaved like the PerL muscle after HSDA with maintained muscle and MU contractile force and speed but with a dramatic increase in fatigability, irrespective of whether all the dorsal roots were transected. We conclude that reduced neuromuscular activity accounts for increased fatigability but is not, in of itself, sufficient to promote atrophy and slow to fast conversion. Position and relative movements of hindlimb muscles are likely contributors to sustained MG muscle and MU contractile force and speed after HSDA and HSDA-SP surgeries.

Myosin Heavy Chain Composition, Rate of Dystrophin and Integrin Degradation and Meat Quality of Pig Longissimus thoracis and psoas major Muscles During Postmortem Aging

Abstract This study evaluated myosin heavy chain composition and the effect of aging time (45 min, 24 h, 96 h and 168 h) on meat quality parameters and dystrophin and integrin degradation pattern in longissimus thoracis and psoas major muscles of 24 Polish Landrace fatteners slaughtered at 100 kg body weight. It was found that m. longissimus thoracis had a greater percentage of MyHC-IIB (P≤0.05) and a significantly lower percentage of type I and IIA/IIX (P≤0.05) compared with the m. psoas major. Moreover, psoas major muscle had significantly higher (P≤0.05) pH45, pH24 and lower drip loss values for 45 min to 24 h, 45 min to 96 h and 45 min to 168 h than m. longissimus thoracis. Muscle type also had a significant effect on the rate of dystrophin and integrin degradation postmortem, which were significantly (P≤0.05) more rapidly degraded in the m. longissimus thoracis compared to the m. psoas major. It is concluded from the obtained results that the muscles with a greater percentage of MyHC-IIB show lower pH values, which translates into more rapid rate of integrin and dystrophin degradation. Ultimately, this may contribute to higher drip loss values during refrigerated storage.