Sarcomere Length Research Articles

The quality and nutritional value of beef from Angus steers fed different levels of humate (K Humate S100R)

This study compared the effect of four levels of K Humate S100R (potassium humate) supplementation on the quality, shelf-life, and nutritional properties of beef. Angus steers (n = 40) were individually housed and fed either 0, 35, 70, or 140 g K Humate S100R/animal/day for 100 days, following a 30 day adjustment period. The steers were slaughtered at the completion of the feeding study. The left m. longissimus lumborum (LL) was collected at 24 h post-mortem and aged for either 2 or 6 weeks before analysis. K Humate S100R supplementation did not affect beef drip loss, cooking loss, shear force, sarcomere length, ultimate pH, intramuscular fat content, or total volatile basic nitrogen concentrations (P > 0.05). Steers supplemented with 70 g/day K Humate S100R produced beef with higher a* values on Days 1 and 3 of retail display (P < 0.05). Beef mineral composition was unchanged by K Humate S100R supplementation (P > 0.05), but there were minor changes to the fatty acid profile. Specifically, the ratio of omega-6 to omega-3 (P < 0.05) and C20:2n-6 concentrations (P < 0.05) increased with supplementation level. Together, these results demonstrate no detrimental effects on beef quality and shelf-life as a result of K Humate S100R supplementation.

Stretch-shortening cycles protect against the age-related loss of power generation in rat single muscle fibres

Aging is associated with impaired strength and power during isometric and shortening contractions, however, during lengthening (i.e., eccentric) contractions, strength is maintained. During daily movements, muscles undergo stretch-shortening cycles (SSCs). It is unclear whether the age-related maintenance of eccentric strength offsets age-related impairments in power generation during SSCs owing to the utilization of elastic energy or other cross-bridge based mechanisms. Here we investigated how aging influences SSC performance at the single muscle fibre level and whether performing active lengthening prior to shortening protects against age-related impairments in power generation. Single muscle fibres from the psoas major of young (∼8 months; n = 31 fibres) and old (∼32 months; n = 41 fibres) male F344BN rats were dissected and chemically permeabilized. Fibres were mounted between a force transducer and length controller and maximally activated (pCa 4.5). For SSCs, fibres were lengthened from average sarcomere lengths of 2.5 to 3.0 μm and immediately shortened back to 2.5 μm at both fast and slow (0.15 and 0.60 Lo/s) lengthening and shortening speeds. The magnitude of the SSC effect was calculated by comparing work and power during shortening to an active shortening contraction not preceded by active lengthening. Absolute isometric force was ∼37 % lower in old compared to young rat single muscle fibres, however, when normalized to cross-sectional area (CSA), there was no longer a significant difference in isometric force between age groups, meanwhile there was an ∼50 % reduction in absolute power in old as compared with young. We demonstrated that SSCs significantly increased power production (75–110 %) in both young and old fibres when shortening occurred at a fast speed and provided protection against power-loss with aging. Therefore, in older adults during everyday movements, power is likely ‘protected’ in part due to the stretch-shortening cycle as compared with isolated shortening contractions.

Effects of Neuromuscular Electrical Stimulation and Therapeutic Ultrasound on Quadriceps Contracture of Immobilized Rats.

Quadriceps contracture is a condition where the muscle-tendon unit is abnormally shortened. The treatment prognosis is guarded to poor depending on the progress of the disease. To improve the prognosis, we investigated the effectiveness of therapeutic ultrasound and NMES in treating quadriceps contracture in an immobilized rat model. Thirty-six Wistar rats were randomized into control, immobilization alone, immobilization and spontaneous recovery, immobilization and therapeutic ultrasound, immobilization and NMES, and immobilization and therapeutic ultrasound and NMES combination groups. The continuous therapeutic ultrasound (frequency, 3 MHz, intensity 1 W/cm2) and NMES (TENS mode, frequency 50 Hz; intensity 5.0 ± 0.8 mA) were performed on the quadriceps muscle. On Day 15, immobilization-induced quadriceps contracture resulted in a decreased ROM of the stifle joint, reduction in the sarcomere length, muscle atrophy, and muscle fibrosis. On Day 43, therapeutic ultrasound, NMES, and combining both methods improved muscle atrophy and shortening and decreased collagen type I and III and α-SMA protein. The combination of therapeutic ultrasound and NMES significantly reduced the mRNA expression of IL-1β, TGF-β1, and HIF-1α and increased TGF-β3. Therefore, the combination of therapeutic ultrasound and NMES is the most potent rehabilitation program for treating quadriceps contracture.

Comparative analysis of ventricular stiffness across species.

Investigating ventricular diastolic properties is crucial for understanding the physiological cardiac functions in organisms and unraveling the pathological mechanisms of cardiovascular disorders. Ventricular stiffness, a fundamental parameter that defines ventricular diastolic functions in chordates, is typically analyzed using the end-diastolic pressure-volume relationship (EDPVR). However, comparing ventricular stiffness accurately across chambers of varying maximum volume capacities has been a long-standing challenge. As one of the solutions to this problem, we propose calculating a relative ventricular stiffness index by applying an exponential approximation formula to the EDPVR plot data of the relationship between ventricular pressure and values of normalized ventricular volume by the ventricular weight. This article reviews the potential, utility, and limitations of using normalized EDPVR analysis in recent studies. Herein, we measured and ranked ventricular stiffness in differently sized and shaped chambers using exvivo ventricular pressure-volume analysis data from four animals: Wistar rats, red-eared slider turtles, masu salmon, and cherry salmon. Furthermore, we have discussed the mechanical effects of intracellular and extracellular viscoelastic components, Titin (Connectin) filaments, collagens, physiological sarcomere length, and other factors that govern ventricular stiffness. Our review provides insights into the comparison of ventricular stiffness in different-sized ventricles between heterologous and homologous species, including non-model organisms.

Effects of Sous-vide Cooking Temperature on Triceps Brachii of Black Goats.

The aim of this study was to determine the effects of sous-vide cooking temperature on the triceps brachii of black goats. Triceps brachii of black goats (12 months) were sous-vide cooked at 55°C, 60°C, and 65°C. The samples were examined for color, scanning electron microscope photographs, sarcomere length, fiber cross-sectional area, cooking yield, shear force, sensory evaluation, and aromatic profile. The results showed that CIE a*, CIE b*, and chroma increased with increasing sous-vide cooking temperature. However, the cooking yield significantly decreased with increasing sous-vide cooking temperature, and the shear forces of the 60°C and 65°C samples showed no significant differences. For sensory evaluation, the 60°C sample showed the highest scores for flavor, texture, and off-flavor. Furthermore, the 60°C sample showed the significantly lowest value of octadienone (aroma characteristics of metallic) intensity (p<0.05). Therefore, sous-vide cooking of triceps brachii of black goats at 60°C is effective in reducing off-flavor and improving tenderness.

Effect of Electrostatic Field Assisted Thawing on the Quality of Previously Frozen Beef Striploins

The objective of this study was to evaluate the impact of applying electrostatic field (EF)–assisted thawing on the quality attributes of previously frozen beef striploin. Beef striploins from both sides of 12 USDA Choice carcasses were halved, frozen at −40°C, and thawed under 4 EF voltage treatments: 0 kV (control), 2.5 kV, 5 kV, and 10 kV. After reaching the internal temperature of −1°C, striploins were weighed for yield calculation, swabbed for microbial analysis, fabricated into steaks, and assigned to either 0- or 14-d aging period and retail displayed for 0 or 7 d. Subjective and objective color measurements were taken during the retail display. Upon completion of retail display, Warner-Bratzler shear force (WBSF), cook loss, sarcomere length, troponin-T degradation, muscle fiber spacing, lipid oxidation, antioxidant capacity, pH, and proximate analysis were performed. All EF treatments increased purge loss compared to the control (P&amp;lt;0.05) and did not improve thawing speed, with samples from 10 kV actually taking the longest to thaw (P&amp;lt;0.05). The 2.5 kV and 5 kV samples aged 14 d showed less discoloration than those from 0 kV and 10 kV, and 5 kV samples aged 14 d had higher a* than those from the other treatments (P&amp;lt;0.05). Samples thawed under 10 kV showed a reduction in WBSF compared to the control (P&amp;lt;0.05), but there was no impact of EF on aerobic plate count, sarcomere length, troponin-T degradation,relative fat %, crude protein %, moisture %, purge protein concentration, pH, lipid oxidation, or antioxidant capacity for either the hydrophilic (water soluble) and lipophilic (lipid soluble) portion of the samples (P&amp;gt;0.05). Overall, our study determined that there was no economic benefit to apply EF during thawing regarding yield and purge loss. However, the application of EF may improve tenderness and extend shelf life of beef during retail display.

Maternal exercise preserves offspring cardiovascular health via oxidative regulation of the ryanodine receptor

ObjectiveThe intrauterine environment during pregnancy is a critical factor in the development of obesity, diabetes, and cardiovascular disease in offspring. Maternal exercise prevents the detrimental effects of a maternal high fat diet on the metabolic health in adult offspring, but the effects of maternal exercise on offspring cardiovascular health have not been thoroughly investigated. MethodsTo determine the effects of maternal exercise on offspring cardiovascular health, female mice were fed a chow (C; 21% kcal from fat) or high-fat (H; 60% kcal from fat) diet and further subdivided into sedentary (CS, HS) or wheel exercised (CW, HW) prior to pregnancy and throughout gestation. Offspring were maintained in a sedentary state and chow-fed throughout 52 weeks of age and subjected to serial echocardiography and cardiomyocyte isolation for functional and mechanistic studies. ResultsHigh-fat fed sedentary dams (HS) produced female offspring with reduced ejection fraction (EF) compared to offspring from chow-fed dams (CS), but EF was preserved in offspring from high-fat fed exercised dams (HW) throughout 52 weeks of age. Cardiomyocytes from HW female offspring had increased kinetics, calcium cycling, and respiration compared to CS and HS offspring. HS offspring had increased oxidation of the RyR2 in cardiomyocytes coupled with increased baseline sarcomere length, resulting in RyR2 overactivity, which was negated in female HW offspring. ConclusionsThese data suggest a role for maternal exercise to protect against the detrimental effects of a maternal high-fat diet on female offspring cardiac health. Maternal exercise improved female offspring cardiomyocyte contraction, calcium cycling, respiration, RyR2 oxidation, and RyR2 activity. These data present an important, translatable role for maternal exercise to preserve cardiac health of female offspring and provide insight on mechanisms to prevent the transmission of cardiovascular diseases to subsequent generations.

Multivariate Examination of Metabolic Contributions to Beef Longissimus Lumborum Flavor

We examined the impact of muscle metabolic capacity on beef longissimus lumborum flavor. Beef carcasses were selected to have normal or dark cutting lean color (n=160, each) and aged until 13 d postmortem. Muscle pH, glycolytic potential, mitochondrial DNA copy number, instrumental lean color, myoglobin concentration, carbonyls on sarcoplasmic proteins, initial metmyoglobin formation, bloom, proximate composition, slice shear force, sarcomere length, desmin degradation, overall tenderness, juiciness, and flavor profile were determined. Carcasses were clustered based on metabolic characteristics into dark cutting classes (Control, Shady, Moderate, and Severe), which were compared using analysis of variance and multiple factor analysis. Clusters were in general, but not complete, agreement with classifications based on muscle pH. Multiple factor analysis produced 2 dimensions that explained 30.8% and 13.8% of the variation, respectively. Dimension 1 had strong negative loadings for muscle pH and strong positive loadings for glycolytic potential, L*, a*, b*, initial metmyoglobin formation, and bloom. Ratings for fat-like, overall sweet, sweet, and musty/earthy/humus had relatively weak positive loadings for dimension 1, whereas salt, sour, and metallic ratings had weak negative loadings for dimension 1. Overall tenderness and juiciness ratings, marbling score, intramuscular lipid content, carbonyls on sarcoplasmic proteins, and L* had positive loadings for dimension 2. Ratings for fat-like, beef flavor identity, and brown/roasted had positive loadings for dimension 2, and intramuscular moisture content, slice shear force, and mitochondrial copy number had negative loadings. Sample scores stratified dark cutting clusters along dimension 1, which agreed with univariate comparisons for these traits. Sample scores for dimension 2 were greater for Moderate and Control steaks than for Shady steaks. These data indicate that clustering was effective in segmenting them into groupings more indicative of the metabolic machinery than pH alone. Moreover, the semetabolic differences influenced animal variation in beef flavor profile.

Translating myosin-binding protein C and titin abnormalities to whole-heart function using a novel calcium-contraction coupling model

Mutations in cardiac myosin-binding protein C (cMyBP-C) or titin may respectively lead to hypertrophic (HCM) or dilated (DCM) cardiomyopathies. The mechanisms leading to these phenotypes remain unclear because of the challenge of translating cellular abnormalities to whole-heart and system function.We developed and validated a novel computer model of calcium-contraction coupling incorporating the role of cMyBP-C and titin based on the key assumptions: 1) tension in the thick filament promotes cross-bridge attachment mechanochemically, 2) with increasing titin tension, more myosin heads are unlocked for attachment, and 3) cMyBP-C suppresses cross-bridge attachment.Simulated stationary calcium-tension curves, isotonic and isometric contractions, and quick release agreed with experimental data. The model predicted that a loss of cMyBP-C function decreases the steepness of the calcium-tension curve, and that more compliant titin decreases the level of passive and active tension and its dependency on sarcomere length. Integrating this cellular model in the CircAdapt model of the human heart and circulation showed that a loss of cMyBP-C function resulted in HCM-like hemodynamics with higher left ventricular end-diastolic pressures and smaller volumes. More compliant titin led to higher diastolic pressures and ventricular dilation, suggesting DCM-like hemodynamics.The novel model of calcium-contraction coupling incorporates the role of cMyBP-C and titin. Its coupling to whole-heart mechanics translates changes in cellular calcium-contraction coupling to changes in cardiac pump and circulatory function and identifies potential mechanisms by which cMyBP-C and titin abnormalities may develop into HCM and DCM phenotypes. This modeling platform may help identify distinct mechanisms underlying clinical phenotypes in cardiac diseases.

Is Drosophila indirect flight muscle a good model to investigate residual force enhancement with minimal sarcomere length non-uniformities?

Is Drosophila indirect flight muscle a good model to investigate residual force enhancement with minimal sarcomere length non-uniformities?

Simulated microgravity improves maturation of cardiomyocytes derived from human induced pluripotent stem cells

Cardiomyocytes derived from human induced pluripotent stem cells (hiPSC-CMs) possess tremendous potential for basic research and translational application. However, these cells structurally and functionally resemble fetal cardiomyocytes, which is a major limitation of these cells. Microgravity can significantly alter cell behavior and function. Here we investigated the effect of simulated microgravity on hiPSC-CM maturation. Following culture under simulated microgravity in a random positioning machine for 7 days, 3D hiPSC-CMs had increased mitochondrial content as detected by a mitochondrial protein and mitochondrial DNA to nuclear DNA ratio. The cells also had increased mitochondrial membrane potential. Consistently, simulated microgravity increased mitochondrial respiration in 3D hiPSC-CMs, as indicated by higher levels of maximal respiration and ATP content, suggesting improved metabolic maturation in simulated microgravity cultures compared with cultures under normal gravity. Cells from simulated microgravity cultures also had improved Ca2+ transient parameters, a functional characteristic of more mature cardiomyocytes. In addition, these cells had improved structural properties associated with more mature cardiomyocytes, including increased sarcomere length, z-disc length, nuclear diameter, and nuclear eccentricity. These findings indicate that microgravity enhances the maturation of hiPSC-CMs at the structural, metabolic, and functional levels.

Three-dimensional co-culturing of stem cell-derived cardiomyocytes and cardiac fibroblasts reveals a role for both cell types in Marfan-related cardiomyopathy

Pathogenic variants in the FBN1 gene, which encodes the extracellular matrix protein fibrillin-1, cause Marfan syndrome (MFS), which affects multiple organ systems, including the cardiovascular system. Myocardial dysfunction has been observed in a subset of patients with MFS and in several MFS mouse models. However, there is limited understanding of the intrinsic consequences of FBN1 variants on cardiomyocytes (CMs). To elucidate the CM-specific contribution in Marfan's cardiomyopathy, cardiosphere cultures of CMs and cardiac fibroblasts (CFs) are used. CMs and CFs were derived by human induced pluripotent stem cell (iPSC) differentiation from MFS iPSCs with a pathogenic variant in FBN1 (c.3725G>A; p.Cys1242Tyr) and the corresponding CRISPR-corrected iPSC line (Cor).Cardiospheres containing MFS CMs show decreased FBN1, COL1A2 and GJA1 expression. MFS CMs cultured in cardiospheres have fewer binucleated CMs in comparison with Cor CMs. 13% of MFS CMs in cardiospheres are binucleated and 15% and 16% in cardiospheres that contain co-cultures with respectively MFS CFs and Cor CFs, compared to Cor CMs, that revealed up to 23% binucleation when co-cultured with CFs. The sarcomere length of CMs, as a marker of development, is significantly increased in MFS CMs interacting with Cor CF or MFS CF, as compared to monocultured MFS CMs. Nuclear blebbing was significantly more frequent in MFS CFs, which correlated with increased stiffness of the nuclear area compared to Cor CFs.Our cardiosphere model for Marfan-related cardiomyopathy identified a contribution of CFs in Marfan-related cardiomyopathy and suggests that abnormal early development of CMs may play a role in the disease mechanism.

Gaining new understanding of sarcomere length non-uniformities in skeletal muscles.

Sarcomere lengths are non-uniform on all structural levels of mammalian skeletal muscle. These non-uniformities have been associated with a variety of mechanical properties, including residual force enhancement and depression, creep, increased force capacity, and extension of the plateau of the force-length relationship. However, the nature of sarcomere length non-uniformities has not been explored systematically. The purpose of this study was to determine the properties of sarcomere length non-uniformities in active and passive muscle. Single myofibrils of rabbit psoas (n = 20; with 412 individual sarcomeres) were subjected to three activation/deactivation cycles and individual sarcomere lengths were measured at 4 passive and 3 active points during the activation/deactivation cycles. The myofibrils were divided into three groups based on their initial average sarcomere lengths: short, intermediate, and long average sarcomere lengths of 2.7, 3.2, and 3.6µm. The primary results were that sarcomere length non-uniformities did not occur randomly but were governed by some structural and/or contractile properties of the sarcomeres and that sarcomere length non-uniformities increased when myofibrils went from the passive to the active state. We propose that the mechanisms that govern the systematic sarcomere lengths non-uniformities observed in active and passive myofibrils may be associated with the variable number of contractile proteins and the variable number and the adjustable stiffness of titin filaments in individual sarcomeres.

Peripheral thickening of the sarcomeres and pointed end elongation of the thin filaments are both promoted by SALS and its formin interaction partners.

During striated muscle development the first periodically repeated units appear in the premyofibrils, consisting of immature sarcomeres that must undergo a substantial growth both in length and width, to reach their final size. Here we report that, beyond its well established role in sarcomere elongation, the Sarcomere length short (SALS) protein is involved in Z-disc formation and peripheral growth of the sarcomeres. Our protein localization data and loss-of-function studies in the Drosophila indirect flight muscle strongly suggest that radial growth of the sarcomeres is initiated at the Z-disc. As to thin filament elongation, we used a powerful nanoscopy approach to reveal that SALS is subject to a major conformational change during sarcomere development, which might be critical to stop pointed end elongation in the adult muscles. In addition, we demonstrate that the roles of SALS in sarcomere elongation and radial growth are both dependent on formin type of actin assembly factors. Unexpectedly, when SALS is present in excess amounts, it promotes the formation of actin aggregates highly resembling the ones described in nemaline myopathy patients. Collectively, these findings helped to shed light on the complex mechanisms of SALS during the coordinated elongation and thickening of the sarcomeres, and resulted in the discovery of a potential nemaline myopathy model, suitable for the identification of genetic and small molecule inhibitors.

Lactate- and immunomagnetic-purified hiPSC-derived cardiomyocytes generate comparable engineered cardiac tissue constructs.

Three-dimensional engineered cardiac tissue (ECT) using purified human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) has emerged as an appealing model system for the study of human cardiac biology and disease. A recent study reported widely used metabolic (lactate) purification of monolayer hiPSC-CM cultures results in an ischemic cardiomyopathy-like phenotype compared with magnetic antibody-based cell sorting (MACS) purification, complicating the interpretation of studies using lactate-purified hiPSC-CMs. Herein, our objective was to determine if use of lactate relative to MACS-purified hiPSC-CMs affects the properties of resulting hiPSC-ECTs. Therefore, hiPSC-CMs were differentiated and purified using either lactate-based media or MACS. Global proteomics revealed that lactate-purified hiPSC-CMs displayed a differential phenotype over MACS hiPSC-CMs. hiPSC-CMs were then integrated into 3D hiPSC-ECTs and cultured for 4 weeks. Structurally, there was no significant difference in sarcomere length between lactate and MACS hiPSC-ECTs. Assessment of isometric twitch force and Ca2+ transient measurements revealed similar functional performance between purification methods. High-resolution mass spectrometry-based quantitative proteomics showed no significant difference in protein pathway expression or myofilament proteoforms. Taken together, this study demonstrates that lactate- and MACS-purified hiPSC-CMs generate ECTs with comparable structural, functional, and proteomic features, and it suggests that lactate purification does not result in an irreversible change in a hiPSC-CM phenotype.

Age and dietary restriction modulate mitochondrial quality in quadriceps femoris muscle of male mice.

Dietary restriction (DR) is a potential intervention for ameliorating ageing-related damages. Mitochondrial quality control is the key mechanism for regulating cellular functions in skeletal muscle. This study aimed to explore the effect of age and DR on the homeostasis of mitochondrial quality control in skeletal muscle. To study the effect of age on mitochondrial homeostasis, young (3 months old) male C57BL/6J mice were fed ad libitum (AL) until 7 (Young), 14 (Middle), and 19 months (Aged) of age. For the DR intervention, 60% of AL intake was given to the mice at 3 months of age until they reached 19 months of age (16 months). The quadriceps femoris muscle was collected for further analysis. Significant changes in the skeletal muscle were noticed during the transition between middle age and the elderly stages. An accumulation of collagen was observed in the muscle after middle age. Compared with the Middle muscle, Aged muscle displayed a greater expression of VDAC, and lower expressions of mitochondrial dynamic proteins and OXPHOS proteins. The DR intervention attenuated collagen content and elongated the sarcomere length in the skeletal muscle during ageing. In addition, DR adjusted the abnormalities in mitochondrial morphology in the Aged muscle. DR downregulated VDAC expression, but upregulated OPA1 and DRP1 expressions. Taken together, greater pathological changes were noticed in the skeletal muscle during ageing, especially in the transition between middle age and the elderly, whereas early-onset DR attenuated the muscular ageing via normalising partial functions of mitochondria.

TRPC5 channel participates in myocardial injury in chronic intermittent hypoxia

ObjectiveThe purpose of this study is to develop an animal model of Chronic Intermittent Hypoxia (CIH) and investigate the role of the TRPC5 channel in cardiac damage in OSAHS rats. MethodsTwelve male Sprague Dawley rats were randomly divided into the CIH group and the Normoxic Control (NC) group. Changes in structure, function, and pathology of heart tissue were observed through echocardiography, transmission electron microscopy, HE-staining, and TUNEL staining. ResultsThe Interventricular Septum thickness at diastole (IVSd) and End-Diastolic Volume (EDV) of rats in the CIH group significantly increased, whereas the LV ejection fraction and LV fraction shortening significantly decreased. TEM showed that the myofilaments in the CIH group were loosely arranged, the sarcomere length varied, the cell matrix dissolved, the mitochondrial cristae were partly flocculent, the mitochondrial outer membrane dissolved and disappeared, and some mitochondria were swollen and vacuolated. The histopathological examination showed that the cardiomyocytes in the CIH group were swollen with granular degeneration, some of the myocardial fibers were broken and disorganized, and most of the nuclei were vacuolar and hypochromic. ConclusionCIH promoted oxidative stress, the influx of Ca2+, and the activation of the CaN/NFATc signaling pathway, which led to pathological changes in the morphology and ultrastructure of cardiomyocytes, the increase of myocardial apoptosis, and the decrease of myocardial contractility. These changes may be associated with the upregulation of TRPC5.

Sarcomere level mechanics of the fast skeletal muscle of the medaka fish larva.

The medaka fish (Oryzias latipes) is a vertebrate model used in developmental biology and genetics. Here we explore its suitability as a model for investigating the molecular mechanisms of human myopathies caused by mutations in sarcomeric proteins. To this end, the relevant mechanical parameters of the intact skeletal muscle of wild-type medaka are determined using the transparent tail at larval stage 40. Tails were mounted at sarcomere length of 2.1 μm in a thermoregulated trough containing physiological solution. Tetanic contractions were elicited at physiological temperature (10°C-30°C) by electrical stimulation, and sarcomere length changes were recorded with nanometer-microsecond resolution during both isometric and isotonic contractions with a striation follower. The force output has been normalized for the actual fraction of the cross section of the tail occupied by the myofilament lattice, as established with transmission electron microscopy (TEM), and then for the actual density of myofilaments, as established with X-ray diffraction. Under these conditions, the mechanical performance of the contracting muscle of the wild-type larva can be defined at the level of the half-thick filament, where ∼300 myosin motors work in parallel as a collective motor, allowing a detailed comparison with the established performance of the skeletal muscle of different vertebrates. The results of this study point out that the medaka fish larva is a suitable model for the investigation of the genotype/phenotype correlations and therapeutic possibilities in skeletal muscle diseases caused by mutations in sarcomeric proteins.NEW & NOTEWORTHY The suitability of the medaka fish as a model for investigating the molecular mechanisms of human myopathies caused by mutations of sarcomeric proteins is tested by combining structural analysis and sarcomere-level mechanics of the skeletal muscle of the tail of medaka larva. The mechanical performance of the medaka muscle, scaled at the level of the myosin-containing thick filament, together with its reduced genome duplication makes this model unique for investigations of the genotype/phenotype correlations in human myopathies.

Potentiation of force by extracellular potassium is not dependent on muscle length in mouse EDL muscle.

Increases in myofiber extracellular potassium with prolonged contractile activity can potentiate twitch force. Activity-dependent potentiation, another mechanism of force increase in skeletal muscle, has a strong dependence on muscle or sarcomere length. Thus, potassium-mediated twitch potentiation could also be length-dependent. However, this has not been previously investigated. To this end, we used isolated C57BL/6 mouse extensor digitorum longus (EDL) muscles and elicited twitches at 0.9 Lo, Lo, and 1.1 Lo (Lo refers to optimal length) in normal (5 mM) and high (10 mM) potassium solutions. Potentiation magnitude was similar to previous observations and was not significantly different between lengths (0.9 Lo: 12.3 ± 4.4%, Lo: 12.2 ± 3.6%, 1.1 Lo: 11.8 ± 4.8%, values are means ± SD). Exposure to dantrolene sodium, a compound that attenuates calcium release, reduced twitch force across lengths by ∼70%. When dantrolene-affected muscles were subsequently exposed to high potassium, potentiation was similar to that observed in the absence of the former. In total, these findings provide novel information on potassium-mediated twitch potentiation.NEW & NOTEWORTHY Here, we investigated the length-dependence of twitch force potentiation by extracellular potassium in mouse extensor digitorum longus (EDL) in vitro, at 25°C. Potentiation magnitude did not display a statistically significant difference between the examined muscle lengths. These results describe, for the first time, the relationship of this form of potentiation with muscle length, thus furthering the understanding of how it is integrated in in vivo muscle function.

Hole behavior captured by analysis of instantaneous amplitude and phase of sarcosynced oscillations reveals wave characteristics of sarcomeric oscillations

In this study, we performed signal analysis based on instantaneous amplitude and phase of sarcomeric oscillations, which are generated by skeletal muscle under constant calcium concentration conditions and in which sarcomeres repeatedly contract and relax autonomously. In addition to the changes in sarcomere length that have been attracting attention, we named the Z-line oscillations that partition sarcomeres sarcosynced oscillations, and analyzed their instantaneous amplitude and phase. As a result, the behavior of pairs of sarcosynced oscillations and sarcomeric oscillations, which are produced when propagating waves propagate in one direction or collide, was clearly visualized. By focusing on the behavior of the hole, which is a dip in the instantaneous amplitude accompanied by a sudden jump in the instantaneous phase in sarcosynced oscillations, we were able to discern the wave characteristics. Transient disruption occurred in the propagating waves even when they traveled in one direction. Its properties were captured by the sarcomeric defect hole (SD hole), a dip in the instantaneous amplitude accompanied by a jump in the instantaneous phase in sarcosynced oscillations. When propagating waves collide, the collision site, its persistence, movement, and disappearance process are captured as sarcomeric collision holes (SC holes) of sarcosynced oscillations. These holes are important indicators for understanding the oscillation properties of sarcomeres. In conclusion, although sarcosynced oscillations and sarcomeric oscillations are closely related, they exhibit different oscillations, and the study of the SD holes and SC holes caused by them will contribute to a detailed understanding of the muscle characteristics of sarcomeres. This finding has important implications for improving our understanding of the efficiency of muscle function and its regulatory mechanisms.