Twitch Contraction Research Articles

Advances in Cholinesterase Inhibitor Research-An Overview of Preclinical Studies of Selected Organoruthenium(II) Complexes.

Cholinesterase (ChE) inhibitors are crucial therapeutic agents for the symptomatic treatment of certain chronic neurodegenerative diseases linked to functional disorders of the cholinergic system. Significant research efforts have been made to develop novel derivatives of classical ChE inhibitors and ChE inhibitors with novel scaffolds. Over the past decade, ruthenium complexes have emerged as promising novel therapeutic alternatives for the treatment of neurodegenerative diseases. Our research group has investigated a number of newly synthesized organoruthenium(II) complexes for their inhibitory activity against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Three complexes (C1a, C1-C, and C1) inhibit ChE in a pharmacologically relevant range. C1a reversibly inhibits AChE and BChE without undesirable peripheral effects, making it a promising candidate for the treatment of Alzheimer's disease. C1-Cl complex reversibly and competitively inhibits ChEs, particularly AChE. It inhibits nerve-evoked skeletal muscle twitch and tetanic contraction in a concentration-dependent manner with no effect on directly elicited twitch and tetanic contraction and is promising for further preclinical studies as a competitive neuromuscular blocking agent. C1 is a selective, competitive, and reversible inhibitor of BChE that inhibits horse serum BChE (hsBChE) without significant effect on the peripheral neuromuscular system and is a highly species-specific inhibitor of hsBChE that could serve as a species-specific drug target. This research contributes to the expanding knowledge of ChE inhibitors based on ruthenium complexes and highlights their potential as promising therapeutic candidates for chronic neurodegenerative diseases.

Effects of intensified training with insufficient recovery on joint level and single muscle fibre mechanical function: the role of myofibrillar Ca2+ sensitivity.

Intense exercise training with insufficient recovery time is associated with reductions in neuromuscular performance. However, it is unclear how single muscle fibre mechanical function and myofibrillar Ca2+ sensitivity contribute to these impairments. We investigated the effects of overload training on joint-level neuromuscular performance and cellular-level mechanical function. Fourteen athletes (4 female and 10 male) underwent a 3-week intensified training protocol consisting of up to 150% of their regular training hours with three additional high-intensity training sessions per week. Neuromuscular performance of the knee extensors was assessed via maximal voluntary contraction (MVC) force, electrically evoked twitch contractions, and a force-frequency relationship. Muscle biopsies were taken from the vastus lateralis to assess single fibre mechanical function. Neither MVC force nor twitch parameters were altered following training (all p>0.05), but a rightward shift in the force-frequency curve was observed with average reduction in force of 6%-27% across frequencies 5-20Hz (all p<0.05). In single fibres, maximal force output was not reduced following training, but there was a rightward shift in the force-pCa curve driven by a 6% reduction in Ca2+ sensitivity (p<0.05). These data indicate intensified training leads to impaired Ca2+ sensitivity at the single fibre level, which in part explains impaired neuromuscular function at the joint level during lower frequencies of activation. This is an important consideration for athletes, as performance is often assessed at maximal levels of activation, and these underlying impairments in force generation may be less obvious.

Reliability of Quadriceps Twitch Muscle Properties and Explosive Voluntary Contractions at Different Knee Joint Angles

ABSTRACT Background Measures that assess muscle strength and its development, either voluntarily or involuntarily, are important in the clinical and research context. The main aim of this study was to verify the interday reliability and the minimum detectable change (MDC) of the knee extensors muscles torque using evoked contractions and explosive voluntary contractions (EVC), across six different joint angles. Methods 20 participants, 16 men and 4 women (26.5 ± 6.6 years; 174.5 ± 11.1 cm; 73.7 ± 11.3 kg), participated in this study. Evoked twitch contractions and voluntary explosive contractions of the knee extensors were performed in 2 days (test-retest) at 20, 40, 70, 90, 105 and 115 degrees. Results The most reliable twitch parameters were peak torque (PT), contraction time (CT), half relaxation-time (½ time), total impulse and peak rate of torque development (RTDPeak), with coefficient of variation (CV) ranging from ~ 8% at 20° to 5% at 115° angle. It was verified that CT and ½ time become more reliable as the knee angle changed from 20 to 115 degrees, from shorter to longer muscle lengths (7.3 to 2.7% and 8.8% to 4.0%, respectively). The more reliable parameters for voluntary contraction were PT, RTD250 ms, RTDPeak and impulse250 ms (CV from around ~ 8% at 20°/115° and 5% at 70°/90°). For voluntary contractions, all measures performed at 70° angle were more reliable compared to the other angles. Conclusion It was concluded that for the knee extensor muscles, evoked twitch contractions presented greater reliability across knee angles, with lower CV values when compared to the voluntary measures for the same time windows. Specifically, for the evoked twitch contractions, angles from 70 to 115 degree proved to be highly reliable. On the other hand, for the voluntary contractions, the central angles (70 and 90 degree) were the most reliable.

Acute Responses to Traditional and Cluster-Set Squat Training With and Without Blood Flow Restriction.

Cornejo-Daza, PJ, Sánchez-Valdepeñas, J, Páez-Maldonado, J, Rodiles-Guerrero, L, Boullosa, D, León-Prados, JA, Wernbom, M, and Pareja-Blanco, F. Acute responses to traditional and cluster-set squat training with and without blood flow restriction. J Strength Cond Res 38(8): 1401-1412, 2024-To compare the acute responses to different set configurations (cluster [CLU] vs. traditional [TRA]) under distinct blood flow conditions (free vs. restricted) in full-squat (SQ). Twenty resistance-trained males performed 4 protocols that differed in the set configuration (TRA: continuous repetitions; vs. CLU: 30 seconds of rest every 2 repetitions) and in the blood flow condition (FF: free-flow; vs. blood flow restriction [BFR]: 50% of arterial occlusion pressure). The relative intensity (60% 1RM), volume (3 sets of 8 repetitions), and resting time (2 minutes) were equated. Mean propulsive force (MPF), velocity (MPV) and power (MPP), and electromyography (EMG) parameters were recorded during each repetition. Tensiomyography (TMG), blood lactate, countermovement jump (CMJ) height, maximal voluntary isometric contraction, in SQ, and movement velocity against the load that elicited a 1 m·s -1 velocity at baseline (V1-load) in SQ were assessed at pre- and post-exercise. The CLU protocols allowed a better maintenance of MPF, MPV, MPP, and EMG median frequency during the exercise compared to TRA (clu-time interaction, p < 0.05). The TRA protocols experienced greater impairments post-exercise in TMG- and EMG-derived variables (clu-time interaction, p < 0.05) and SQ and CMJ performance (clu-time interaction, p = 0.08 and p < 0.05, respectively), as well as higher blood lactate concentrations (clu-time interaction, p < 0.001) than CLU. Moreover, BFR displayed decreases in TMG variables (bfr-time interaction, p < 0.01), but BFR-CLU resulted in the greatest reduction in twitch contraction time ( p < 0.001). Cluster sets reduced fatigue during and after the training session and BFR exacerbated impairments in muscle mechanical properties; however, the combination of both could improve contraction speed after exercise.

NaV1.1 and NaV1.6 Play Unique and Essential Roles in Muscle Spindle Afferent Function in Adult Mice

Voltage gated sodium channels (NaVs) are crucial for the generation and propagation of action potentials. There are 9 different isoforms found in mammals (NaV1.1-1.9), and NaV1.1, NaV1.6, and NaV1.7 are found in the muscle proprioceptors that sense muscle stretch, the muscle spindle afferents. This sensory information is critical for motor reflexes and the sense of body and limb position in space. We previously found that mice lacking one or both copies of NaV1.1 in peripheral sensory neurons displayed motor deficits, including intention tremors. Muscle spindle afferents in these mice also had unstable static stretch sensitivity but normal dynamic sensitivity, suggesting a specific deficit in the maintenance of proprioceptor excitability when the Piezo2 channel was likely to be closed (Espino, et al., 2022). We hypothesized that NaV1.6 would also be necessary for normal motor behavior and muscle spindle afferent function. We generated sensory neuron specific deletions of NaV1.6 by crossing NaV1.6fl/fl mice with mice expressing PirtCre (NaV1.6cko) and observed severe motor deficits on the open field and rotarod test only in NaV1.6cko (n=15) and not NaV1.6Het (n=12) mice compared to NaV1.6fl/fl controls (n=8). In contrast to mice lacking NaV1.1, we did not observe intention tremors. We used an ex vivo muscle nerve preparation consisting of the extensor digitorum longus muscle and peroneal branch of the sciatic nerve. The muscle was maintained at the length of maximal twitch contraction (Lo) and given a battery of 9 ramp-and-hold stretches (2.5, 5, and 7.5% Lo repeated 3 times each) and 16 sinusoidal vibrations (5-100 μm amplitude; 10-100 Hz frequency). Preliminary results include recordings from 18 adult mice of both sexes (NaV1.6fl/fl n=8; NaV1.6cko n=10). We were only able to find stretch sensitive responses in 3 of 10 NaV1.6cko mice, and those were severely impaired with firing rates less than 5 Hz. In contrast, we found robust slowly adapting responses to stretch in all 8 NaV1.6fl/fl mice. Only 1 of 10 NaV1.6cko mice had any firing in response to vibration, in contrast to the normal dynamic sensitivity observed in NaV1.6fl/fl and NaV1.1cko afferents. This suggests that NaV1.6 plays a critical role in initiating muscle spindle afferent transmission, whereas NaV1.1 is essential for maintaining proprioceptor excitability during static stretch. Future studies will determine whether loss of NaV1.7, which does not result in motor deficits, alters muscle spindle afferent function. This work was supported by NIH Grant 5SC3GM127195 (KAW) a RISE Fellowship 5R25GM71381 (SO), NIH 5T32GM099608-10 and 1T32GM1144303-01A1 (CE). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Spasmolytic Effect of Flopropione Does Not Involve Catechol-O- methyltransferase (COMT) Inhibition.

Flopropione (Flo) has been used for gallstone and urolithiasis as a spasmolytic agent almost exclusively in Japan. According to the package insert, its main mechanism is catechol-O-methyltransferase (COMT) inhibition and anti-serotonergic effect. This is obviously contrary to pharmacological common sense, but it is described that way in pharmacology textbooks and occurs in questions in the National Examination for Pharmacists in Japan. As this is a serious problem in education, we re-examined the action of Flo. The guinea pig ureter was hardly contracted by serotonin, but noradrenaline (NA) elicited repetitive twitch contraction, which was inhibited by Flo. The sphincter of Oddi (SO) exhibited a spontaneous repetitive twitch contraction, which was inhibited by NA and Flo. The inhibitory effect of NA was reversed by α- and β-blockers, whereas that of Flo was not. Entacapone, a representative COMT inhibitor, did not affect the movement of the ureter and the SO. Nifedipine suppressed carbachol-induced contraction of the taenia coli, spontaneous movement of the SO, and NA-induced contraction of the ureter to almost the same extent, whereas Flo did not inhibit the taenia coli, but inhibited the contraction of the SO and the ureter. The inhibitory pattern of Flo resembled that of the ryanodine receptor agonist 4-chloro-m-cresol and the inositol 1,4,5-trisphosphate (IP3) receptor antagonist 2-aminoethoxydiphenyl borate. It is concluded that COMT inhibition or serotonin inhibition is not involved in the spasmolytic action of Flo. Flo might act on ryanodine receptors and/or IP3 receptors, which are responsible for periodic Ca release from Ca stores, to disrupt coordinated Ca dynamics.

Belt electrode tetanus muscle stimulation reduces denervation-induced atrophy of rat multiple skeletal muscle groups

Belt electrode-skeletal muscle electrical stimulation (B-SES) involves the use of belt-shaped electrodes to contract multiple muscle groups simultaneously. Twitch contractions have been demonstrated to protect against denervation-induced muscle atrophy in rats, possibly through mitochondrial biosynthesis. This study examined whether inducing tetanus contractions with B-SES suppresses muscle atrophy and identified the underlying molecular mechanisms. We evaluated the effects of acute (60 Hz, 5 min) and chronic (60 Hz, 5 min, every alternate day for one week) B-SES on the tibialis anterior (TA) and gastrocnemius (GAS) muscles in Sprague–Dawley rats using belt electrodes attached to both ankle joints. After acute stimulation, a significant decrease in the glycogen content was observed in the left and right TA and GAS, suggesting that B-SES causes simultaneous contractions in multiple muscle groups. B-SES enhanced p70S6K phosphorylation, an indicator of the mechanistic target of rapamycin complex 1 activity. During chronic stimulations, rats were divided into control (CONT), denervation-induced atrophy (DEN), and DEN + electrically stimulated with B-SES (DEN + ES) groups. After seven days of treatment, the wet weight (n = 8–11 for each group) and muscle fiber cross-sectional area (CSA, n = 6 for each group) of the TA and GAS muscles were reduced in the DEN and DEN + ES groups compared with that in the CON group. The DEN + ES group showed significantly higher muscle weight and CSA than those in the DEN group. Although RNA-seq and pathway analysis suggested that mitochondrial biogenesis is a critical event in this phenomenon, mitochondrial content showed no difference. In contrast, ribosomal RNA 28S and 18S (n = 6) levels in the DEN + ES group were higher than those in the DEN group, even though RNA-seq showed that the ribosome biogenesis pathway was reduced by electrical stimulation. The mRNA levels of the muscle proteolytic molecules atrogin-1 and MuRF1 were significantly higher in DEN than those in CONT. However, they were more suppressed in DEN + ES than those in DEN. In conclusion, tetanic electrical stimulation of both ankles using belt electrodes effectively reduced denervation-induced atrophy in multiple muscle groups. Furthermore, ribosomal biosynthesis plays a vital role in this phenomenon.

Simulated microgravity attenuates myogenesis and contractile function of 3D engineered skeletal muscle tissues

While the effects of microgravity on inducing skeletal muscle atrophy have been extensively studied, the impacts of microgravity on myogenesis and its mechanisms remain unclear. In this study, we developed a microphysiological system of engineered muscle tissue (EMT) fabricated using a collagen / Matrigel composite hydrogel and murine skeletal myoblasts. This 3D EMT model allows non-invasive quantitative assessment of contractile function. After applying a 7-day differentiation protocol to induce myotube formation, the EMTs clearly exhibited sarcomerogenesis, myofilament formation, and synchronous twitch and tetanic contractions with electrical stimuli. Using this 3D EMT system, we investigated the effects of simulated microgravity at 10−3 G on myogenesis and contractile function utilizing a random positioning machine. EMTs cultured for 5 days in simulated microgravity exhibited significantly reduced contractile forces, myofiber size, and differential expression of muscle contractile, myogenesis regulatory, and mitochondrial biogenesis-related proteins. These results indicate simulated microgravity attenuates myogenesis, resulting in impaired muscle function.

In silico and in vitro models reveal the molecular mechanisms of hypocontractility caused by TPM1 M8R.

Dilated cardiomyopathy (DCM) is an inherited disorder often leading to severe heart failure. Linkage studies in affected families have revealed hundreds of different mutations that can cause DCM, with most occurring in genes associated with the cardiac sarcomere. We have developed an investigational pipeline for discovering mechanistic genotype-phenotype relationships in DCM and here apply it to the DCM-linked tropomyosin mutation TPM1 M8R. Atomistic simulations predict that M8R increases flexibility of the tropomyosin chain and enhances affinity for the blocked or inactive state of tropomyosin on actin. Applying these molecular effects to a Markov model of the cardiac thin filament reproduced the shifts in Ca2+sensitivity, maximum force, and a qualitative drop in cooperativity that were observed in an in vitro system containing TPM1 M8R. The model was then used to simulate the impact of M8R expression on twitch contractions of intact cardiac muscle, predicting that M8R would reduce peak force and duration of contraction in a dose-dependent manner. To evaluate this prediction, TPM1 M8R was expressed via adenovirus in human engineered heart tissues and isometric twitch force was observed. The mutant tissues manifested depressed contractility and twitch duration that agreed in detail with model predictions. Additional exploratory simulations suggest that M8R-mediated alterations in tropomyosin-actin interactions contribute more potently than tropomyosin chain stiffness to cardiac twitch dysfunction, and presumably to the ultimate manifestation of DCM. This study is an example of the growing potential for successful in silico prediction of mutation pathogenicity for inherited cardiac muscle disorders.

Effects of acute phosphodiesterase type 5 inhibition on skeletal muscle interstitial PO2 during contractions and recovery

The oxygen partial pressure within the interstitial space (PO2is; mmHg) provides the driving force for oxygen diffusion into the myocyte thereby supporting oxidative phosphorylation. We tested the hypothesis that potentiation of the nitric oxide pathway with sildenafil (phosphodiesterase type 5 inhibitor) would enhance PO2is during muscle metabolic transitions, thereby slowing PO2is on- and accelerating PO2is off-kinetics. The rat spinotrapezius muscle (n = 17) was exposed for PO2is measurements via phosphorescence quenching under control (CON), low-dose sildenafil (1 mg/kg i.a., SIL1) and high-dose sildenafil (7 mg/kg i.a., SIL7). Data were collected at rest and during submaximal twitch contractions (1 Hz, 4–6 V, 3 min) and recovery (3 min). Mean arterial blood pressure (MAP; mmHg) was reduced with both SIL1 (pre:132 ± 5; post:99 ± 5) and SIL7 (pre:111 ± 6; post:99 ± 4) (p < 0.05). SIL7 elevated resting PO2is (18.4 ± 1.1) relative to both CON (15.7 ± 0.7) and SIL1 (15.2 ± 0.7) (p < 0.05). In addition, SIL7 increased end-recovery PO2is (17.7 ± 1.6) compared to CON (12.8 ± 0.9) and SIL1 (13.4 ± 0.8) (p < 0.05). The overall PO2is response during recovery (i.e., area under the PO2is curve) was greater in SIL7 (4107 ± 444) compared to CON (3493 ± 222) and SIL1 (3114 ± 205 mmHg s) (p < 0.05). Contrary to our hypothesis, there was no impact of acute SIL (1 or 7 mg/kg) on the speed of the PO2is response during contractions or recovery (p > 0.05). However, sildenafil lowered MAP and improved skeletal muscle interstitial oxygenation in healthy rats. Specifically, SIL7 enhanced PO2is at rest and during recovery from submaximal muscle contractions. Potentiation of the nitric oxide pathway with sildenafil enhances microvascular blood-myocyte O2 transport and is expected to improve repeated bouts of contractile activity.

Neuromechanical characterization of the abductor hallucis and its potential role in upright postural control.

There is growing evidence to support a role for the abductor hallucis (AH) in standing balance control; however, functional properties of the muscle that may provide more insight into AH's specific contribution to upright posture have yet to be characterized. This study was conducted to quantify functional neuromechanical properties of the AH and correlate the measures with standing balance variables. We quantified strength and voluntary activation during maximal voluntary isometric contractions of the great toe abductor in nine (3 females and 6 males) healthy, young participants. During electrically evoked twitch and tetanic contractions, we measured great toe abduction peak force and constructed a force-frequency curve. We also evaluated peak abduction force, contraction time (CT), half-relaxation time (HRT), rate of force development (RFD), and relaxation rate (RR) from twitch contractions evoked using doublet stimuli. Strength, VA, CT, HRT, RFD, and RR were correlated to centre of pressure standard deviation (COP SD) and velocity (COP VEL) variables of the traditional COP trace and its rambling and trembling components during single-legged stance. AH twitch properties (e.g., CT: 169.8±32.3ms; HRT: 124.1±29.2ms) and force-frequency curve were similar to other slow contractile muscles. Contractile speed related negatively with COP VEL, suggesting AH may be appropriate for slow, prolonged tasks such as ongoing postural balance control. Correlation coefficient outcomes for all variables were similar between rambling and trembling components. Our results provide further evidence for the importance of AH neuromechanical function for standing balance control, at least during a challenging single-legged posture.

In vivo cytosolic H2O2 changes and Ca2+ homeostasis in mouse skeletal muscle.

Hydrogen peroxide (H2O2) and calcium ions (Ca2+) are functional regulators of skeletal muscle contraction and metabolism. Although H2O2 is one of the activators of the type-1 ryanodine receptor (RyR1) in the Ca2+ release channel, the interdependence between H2O2 and Ca2+ dynamics remains unclear. This study tested the following hypotheses using an in vivo model of mouse tibialis anterior (TA) skeletal muscle. 1. Under resting conditions, elevated cytosolic H2O2 concentration ([H2O2]cyto) leads to a concentration-dependent increase in cytosolic Ca2+concentration ([Ca2+]cyto) through its effect on RyR1. 2. In hypoxia (cardiac arrest) and muscle contractions (electrical stimulation), increased [H2O2]cyto induce Ca2+ accumulation. Cytosolic H2O2 (HyPer7) and Ca2+ (Fura-2) dynamics were resolved by TA bioimaging in C57BL/6J male mice under four conditions: Elevated exogenous H2O2, Cardiac arrest, Twitch and Tetanic contractions. Exogenous H2O2 (0.1-100mM) induced a concentration-dependent increase in [H2O2]cyto (+55%,0.1mM; +280%,100mM) and an increase in [Ca2+]cyto (+3%,1.0mM; +8%,10mM). This increase in [Ca2+]cyto was inhibited by pharmacological inhibition of RyR1 by dantrolene. Cardiac arrest-induced hypoxia increased [H2O2]cyto (+33%) and [Ca2+]cyto (+20%) 50min post-cardiac arrest. Compared to exogenous 1.0mM H2O2 condition, [H2O2]cyto after tetanic contractions rose less than one-tenth as much, while [Ca2+]cyto was 4.7-fold higher. In conclusion, substantial increases in [H2O2]cyto levels evoke only modest Ca2+ accumulation via their effect on the sarcoplasmic reticulum RyR1. On the other hand, contrary to hypoxia secondary to cardiac arrest, increases in [H2O2]cyto from contractions are small, indicating that H2O2 generation is unlikely to be a primary factor driving the significant Ca2+ accumulation after, especially tetanic, muscle contractions.

Weakened Contractile Performance and Mitochondrial Respiratory Complex Activity in Skeletal Muscle Improve during Interbout Arousal in Hibernating Daurian Ground Squirrel, Spermophilus dauricus

Mammalian hibernation is composed of multiple episodes of torpor bout, separated by phases of interbout arousal. During torpor, the skeletal muscles of mammals are undoubtedly inactive, but it has been proven to mitigate disuse atrophy. While interbout arousal has been implicated in the prevention of muscle atrophy, the underlying mechanisms sustaining muscle contraction remain to be explored. In the present study, Daurian ground squirrels (Spermophilus dauricus) were divided into four groups: pre-hibernation (PRE), torpor (TOR), interbout arousal (IBA), and post-hibernation (POST). The contractile performance of slow-twitch soleus muscle (SOL) and fast-twitch extensor digitorum longus muscle (EDL) was detected both in situ and in vitro. Concurrently, mitochondrial respiratory chain complex activity in these muscles was quantified. Our findings revealed that in situ contractile properties of both muscles, including force, power output, time duration, and force development/relaxation rates of twitch contraction, and force and power output of tetanic contraction declined in the TOR group compared to the PRE group, but improved in the IBA and POST groups. Fatigue resistance of muscles, determined by the power output of repetitive tetanic contractions in situ, decreased in the TOR group but recovered in the IBA and POST groups. In vitro studies demonstrated that tetanic contraction power output in isolated muscles increased with muscle temperature in both TOR and IBA groups. However, at the same temperature, power output was consistently lower in the TOR group compared to the IBA group. Moreover, the activity of the mitochondrial respiratory chain complex, especially Complexes I and II, decreased in the TOR group but showed recovery in the IBA and POST groups. These findings suggest that both the contractile performance and fatigue resistance of mammalian skeletal muscle are compromised during torpor but can be improved during interbout arousal and post-hibernation. The rebound in body temperature and rise in mitochondrial respiratory chain complex activity in skeletal muscle are involved in enhancing contractile performance and fatigue resistance. This study suggests that interbout arousal functions as a vital temporal interval during which skeletal muscles can transition from the inactivity induced by torpor to a state of restored contractile functionality. Thus, interbout arousal serves as a behavioral safeguard against disuse-induced damage to skeletal muscles during hibernation.

Effects of exhaustive whole-body exercise and caffeine ingestion on muscle contractile properties in healthy men.

The influence of exhaustive whole-body exercise and caffeine ingestion on electromechanical delay (EMD) has been underexplored. This study investigated the effect of exhaustive cycling exercise on EMD and other parameters of muscle contractile properties and the potential ability of caffeine to attenuate the exercise-induced impairments in EMD and muscle contractile properties. Ten healthy men cycled until exhaustion (88±2% of V̇O<inf>2max</inf>) on two separate days after ingesting caffeine (5 mg.kg-1 of body mass) or cellulose (placebo). Parameters of muscle contractile properties of the quadriceps muscles were assessed via volitional and electrically evoked isometric contractions, performed before and 50 minutes after ingestion of the capsules, and after exercise. Muscle recruitment during volitional contractions was determined via surface electromyography. Exhaustive cycling exercise did not affect volitional and relaxation EMD (P>0.05) but increased evoked EMD. In addition, the exhaustive cycling exercise also increased muscle recruitment at the beginning of volitional isometric muscle contraction (P<0.05). The peak twitch force, maximal rate of twitch force development, and twitch contraction time were all compromised after exhaustive cycling exercise (P<0.05). Acute caffeine ingestion had no effect on muscle contractile properties (P>0.05), except that caffeine increased twitch contraction time at postexercise (P<0.05). Exercise-induced decline in peripheral components of the EMD might be compensated by an increase in the muscle recruitment. In addition, acute caffeine ingestion had minimal influence on exercise-induced changes in muscle contractile proprieties.

Electric field stimulation unmasks a subtle role for T-type calcium channels in regulating lymphatic contraction

We previously identified two isoforms of T-type, voltage-gated calcium (Cav3) channels (Cav3.1, Cav3.2) that are functionally expressed in murine lymphatic muscle cells; however, contractile tests of lymphatic vessels from single and double Cav3 knock-out (DKO) mice, exhibited nearly identical parameters of spontaneous twitch contractions as wild-type (WT) vessels, suggesting that Cav3 channels play no significant role. Here, we considered the possibility that the contribution of Cav3 channels might be too subtle to detect in standard contraction analyses. We compared the sensitivity of lymphatic vessels from WT and Cav3 DKO mice to the L-type calcium channel (Cav1.2) inhibitor nifedipine and found that the latter vessels were significantly more sensitive to inhibition, suggesting that the contribution of Cav3 channels might normally be masked by Cav1.2 channel activity. We hypothesized that shifting the resting membrane potential (Vm) of lymphatic muscle to a more negative voltage might enhance the contribution of Cav3 channels. Because even slight hyperpolarization is known to completely silence spontaneous contractions, we devised a method to evoke nerve-independent, twitch contractions from mouse lymphatic vessels using single, short pulses of electric field stimulation (EFS). TTX was present throughout to block the potential contributions of voltage-gated Na+ channels in perivascular nerves and lymphatic muscle. In WT vessels, EFS evoked single contractions that were comparable in amplitude and degree of entrainment to those occurring spontaneously. When Cav1.2 channels were blocked or deleted, only small residual EFS-evoked contractions (~ 5% of normal amplitude) were present. These residual, EFS-evoked contractions were enhanced (to 10–15%) by the KATP channel activator pinacidil (PIN) but were absent in Cav3 DKO vessels. Our results point to a subtle contribution of Cav3 channels to lymphatic contractions that can be unmasked in the absence of Cav1.2 channel activity and when the resting Vm is more hyperpolarized than normal.

Reproducibility of knee extensor and flexor contraction velocity in healthy men and women assessed using tensiomyography: A registered report.

Tensiomyography measures the radial displacement of a muscle during an electrically evoked twitch contraction. Different concepts to determine the rate of displacement (Vc) from the maximum twitch exist, but information on their reproducibility is scarce. Further, different inter-stimuli intervals during progressive stimulation are used, but the effect of different intervals on Vc is unclear. The first aim of this study was to investigate the within and between-day reliability of the five most frequently used Vc concepts. The second aim was to investigate the effect of different inter-stimuli intervals on Vc. On two consecutive days, we determined Vc of the biceps femoris long head and rectus femoris of twenty-four healthy subjects. The maximum displacement was determined twice within three minutes on day one and a third time 24 h later. Also, on day two, we applied three blocks of ten consecutive stimuli at a constant intensity of 50 mA, separated by 3 min each. Inter-stimuli intervals in randomly ordered blocks were 10 s, 20 s or 30 s, respectively. All Vc concepts displayed good to excellent relative (ICC 0.87-0.99) and generally good absolute within- and between-day reliability for both muscles. Across Vc-concepts, absolute reliability was higher for the rectus femoris (CV% 1.3-7.95%) compared to the biceps femoris (CV% 6.06-15.30%). In both muscles, Vc was generally not affected by different inter-stimuli intervals. For most Vc concepts, repeated stimulation induced an increase regardless of the inter-stimuli interval, but this effect was mainly trivial and small at most. The reproducibility of Vc concepts was generally good but varies between different muscles. A rest interval of 10 s seems preferable to longer intervals for less time required per measurement. Following this initial study, the effect of different inter-stimuli intervals on Vc should be further investigated.

Influence of temperature on twitch potentiation following submaximal voluntary contractions in human plantar flexor muscles.

This study aimed to clarify the influence of temperature on post-activation twitch potentiation, a possible indicator of fast-twitch fiber activation during a preceding voluntary contraction. Ten healthy males immersed their left lower leg in water of different temperatures (cold: 0°C, neutral: 32-35°C, hot: ~43°C) for 20 min each. In each temperature condition, they performed submaximal (10%-50% of maximal voluntary contraction torque measured before water immersion) and maximal plantar flexions. Immediately after each voluntary contraction, twitch contractions were evoked with supramaximal stimulation of the posterior tibial nerve. The magnitude of twitch potentiation, defined as a percent increase in twitch torque following a voluntary contraction, increased with the intensity of the preceding voluntary contraction. The magnitude of twitch potentiation after the maximal voluntary contraction was smaller in Cold than in the other temperature conditions. However, temperature had no influence on the relative magnitude of twitch potentiation following the submaximal contractions. In addition, there was no difference in electromyographic activity between the gastrocnemius and soleus muscles in any temperature conditions. Collectively, the temperature dependence was not observed when using twitch potentiation or electromyographic amplitude as an indicator of fast-twitch fiber activation during brief submaximal voluntary contractions.

Rapid restitution of contractile dysfunction by synthetic copolymers in dystrophin-deficient single live skeletal muscle fibers

Duchenne muscular dystrophy (DMD) is caused by the lack of dystrophin, a cytoskeletal protein essential for the preservation of the structural integrity of the muscle cell membrane. DMD patients develop severe skeletal muscle weakness, degeneration, and early death. We tested here amphiphilic synthetic membrane stabilizers in mdx skeletal muscle fibers (flexor digitorum brevis; FDB) to determine their effectiveness in restoring contractile function in dystrophin-deficient live skeletal muscle fibers. After isolating FDB fibers via enzymatic digestion and trituration from thirty-three adult male mice (9 C57BL10, 24 mdx), these were plated on a laminin-coated coverslip and treated with poloxamer 188 (P188; PEO75-PPO30-PEO75; 8400 g/mol), architecturally inverted triblock (PPO15-PEO200-PPO15, 10,700 g/mol), and diblock (PEO75-PPO16-C4, 4200 g/mol) copolymers. We assessed the twitch kinetics of sarcomere length (SL) and intracellular Ca2+ transient by Fura-2AM by field stimulation (25 V, 0.2 Hz, 25 °C). Twitch contraction peak SL shortening of mdx FDB fibers was markedly depressed to 30% of the dystrophin-replete control FDB fibers from C57BL10 (P < 0.001). Compared to vehicle-treated mdx FDB fibers, copolymer treatment robustly and rapidly restored the twitch peak SL shortening (all P < 0.05) by P188 (15 μM = + 110%, 150 μM = + 220%), diblock (15 μM = + 50%, 150 μM = + 50%), and inverted triblock copolymer (15 μM = + 180%, 150 μM = + 90%). Twitch peak Ca2+ transient from mdx FDB fibers was also depressed compared to C57BL10 FDB fibers (P < 0.001). P188 and inverted triblock copolymer treatment of mdx FDB fibers increased the twitch peak Ca2+ transient (P < 0.001). This study shows synthetic block copolymers with varied architectures can rapidly and highly effectively enhance contractile function in live dystrophin-deficient skeletal muscle fibers.

Sex differences in isolated skeletal muscle force production: a spectrum of responses

Whether sex differences in skeletal muscle microvascular responses differ in response to muscle contraction is currently unknown. To determine vascular responses to muscle contraction in vivo we use intravital microscopy using thin skeletal muscle models of mixed fibre types (i.e. hamster retractor muscle). However, to effectively quantify this relationship in vivo, the initial muscle contraction stimulus must be constant across the sexes and thus, we must first determine whether there are sex differences in skeletal muscle force generation. We hypothesized that sexual dimorphisms in force production would not be apparent in muscles of any fibre type composition. To test this hypothesis, we used skeletal muscles with unique fibre type characteristics and oxidative capacities. Mouse soleus (SOL: primarily type 1 &gt;75%), extensor digitorum longus (EDL: primarily type IIB ~60%), and costal diaphragm (DIA: primarily type IIA ~55%) muscles were isolated from adult male and female CD-1 mice. The retractor muscle (RET: primarily type IIX ~58%) was also isolated from adult male and female Syrian hamsters. Muscles were electrically stimulated in vitro and force production was quantified at optimal length during a force-frequency test (1-120Hz) and twitch (1Hz) contractions. In mice, females generated greater force during the force-frequency test in SOL (10-120Hz) and EDL (30-120Hz) and generated greater twitch forces compared to males (SOL: female 34.7 ± 2.4 mN/mm 2 , n=21 vs male 27.8 ± 2.0 mN/mm 2 , n=17; EDL: female 41.2 ± 2.2 mN/mm 2 , n=35 vs male 32.5 ± 2.1 mN/mm 2 , n=29). Females generated greater force during the force frequency test in DIA at 30Hz only and no significant sex differences were observed in DIA twitch force (female 0.24 ± 0.02 g/mg, n=19 vs male 0.20 ± 0.02 g/mg, n=21). In hamster RET no significant sex differences in force were observed during the force frequency test or during the twitch contraction (female 0.13 ± 0.01 g/mg, n=19 vs male 0.14 ± 0.02 g/mg, n=20). Collectively, these findings demonstrate that sex differences in force production were observed in muscles primarily comprised of slow oxidative (type 1) and fast glycolytic (type IIB) fibres but not in muscles primarily comprised of intermediate fibres (type IIA and type IIX). These data show that on a spectrum of muscle fibres from slow oxidative to fast glycolytic, sex differences are most apparent on the extremes of the spectrum, whereas sexual dimorphisms in the middle of the spectrum are not apparent. This work is supported by NSERC. NSERC This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Slower Calcium Handling Balances Faster Cross-Bridge Cycling in Human MYBPC3 HCM.

The pathogenesis of MYBPC3-associated hypertrophic cardiomyopathy (HCM) is still unresolved. In our HCM patient cohort, a large and well-characterized population carrying the MYBPC3:c772G>A variant (p.Glu258Lys, E258K) provides the unique opportunity to study the basic mechanisms of MYBPC3-HCM with a comprehensive translational approach. We collected clinical and genetic data from 93 HCM patients carrying the MYBPC3:c772G>A variant. Functional perturbations were investigated using different biophysical techniques in left ventricular samples from 4 patients who underwent myectomy for refractory outflow obstruction, compared with samples from non-failing non-hypertrophic surgical patients and healthy donors. Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes and engineered heart tissues (EHTs) were also investigated. Haplotype analysis revealed MYBPC3:c772G>A as a founder mutation in Tuscany. In ventricular myocardium, the mutation leads to reduced cMyBP-C (cardiac myosin binding protein-C) expression, supporting haploinsufficiency as the main primary disease mechanism. Mechanical studies in single myofibrils and permeabilized muscle strips highlighted faster cross-bridge cycling, and higher energy cost of tension generation. A novel approach based on tissue clearing and advanced optical microscopy supported the idea that the sarcomere energetics dysfunction is intrinsically related with the reduction in cMyBP-C. Studies in single cardiomyocytes (native and hiPSC-derived), intact trabeculae and hiPSC-EHTs revealed prolonged action potentials, slower Ca2+ transients and preserved twitch duration, suggesting that the slower excitation-contraction coupling counterbalanced the faster sarcomere kinetics. This conclusion was strengthened by in silico simulations. HCM-related MYBPC3:c772G>A mutation invariably impairs sarcomere energetics and cross-bridge cycling. Compensatory electrophysiological changes (eg, reduced potassium channel expression) appear to preserve twitch contraction parameters, but may expose patients to greater arrhythmic propensity and disease progression. Therapeutic approaches correcting the primary sarcomeric defects may prevent secondary cardiomyocyte remodeling.