Failure Of Excitation-contraction Coupling Research Articles

The Effects of Anchoring a Fatiguing Forearm Flexion Task to a High vs. Low Rating of Perceived Exertion on Torque and Neuromuscular Responses.

Ortega, DG, Housh, TJ, Smith, RW, Arnett, JE, Neltner, TJ, Schmidt, RJ, and Johnson, GO. The effects of anchoring a fatiguing forearm flexion task to a high versus low rating of perceived exertion on torque and neuromuscular responses. J Strength Cond Res 38(5): e219-e225, 2024-This study examined the torque and neuromuscular responses following sustained, isometric, forearm flexion tasks anchored to 2 ratings of perceived exertion (RPE). Nine men (mean ± SD: age = 21.0 ± 2.4 years; height = 179.5 ± 5.1 cm; body mass = 79.6 ± 11.4 kg) completed maximal voluntary isometric contractions (MVIC) before and after sustained, isometric, forearm flexion tasks to failure anchored to RPE = 2 and RPE = 8. The amplitude (AMP) and mean power frequency (MPF) of the electromyographic (EMG) signal were recorded from the biceps brachii. Normalized torque was divided by normalized EMG AMP to calculate neuromuscular efficiency (NME). A dependent t-test was used to assess the mean difference for time to task failure (TTF). Repeated-measures analysis of variances was used to compare mean differences for MVIC and normalized neuromuscular parameters. There was no significant difference in TTF between RPE = 2 and RPE = 8 (p = 0.713). The MVIC decreased from pretest to posttest at RPE = 2 (p = 0.009) and RPE = 8 (p = 0.003), and posttest MVIC at RPE = 8 was less than that at RPE = 2 (p < 0.001). In addition, NME decreased from pretest to posttest (p = 0.008). There was no change in normalized EMG AMP or EMG MPF (p > 0.05). The current findings indicated that torque responses were intensity specific, but TTF and neuromuscular responses were not. Furthermore, normalized EMG AMP and EMG MPF remained unchanged but NME decreased, likely due to peripheral fatigue and excitation-contraction coupling failure. Thus, this study provides information regarding the neuromuscular responses and mechanisms of fatigue associated with tasks anchored to RPE, which adds to the foundational understanding of the relationship between resistance exercise and the perception of fatigue.

Co-existence of peripheral fatigue of the knee extensors and jump potentiation after an incremental running test to exhaustion in endurance trained male runners.

The aim of the present study was to investigate the effect of an incremental running exercise until exhaustion on twitch responses and jump capacity in endurance trained runners. For this purpose, 8 experienced endurance male runners were required to perform neuromuscular function tests before and after a submaximal running bout (control condition -CTR-) or an incremental running test to volitional exhaustion (experimental conditions -EXP-). The twitch interpolation technique was used to assess voluntary activation and muscle contractile properties before and after each condition (CTR and EXP). Countermovement jump was also used to assess the stretch-shortening cycle function before and after both conditions. In addition, rating of perceived exertion, heart rate, blood lactate and skin temperature were also recorded. Only EXP improved jump performance, however, it was also accompanied by a reduction in maximal voluntary contraction and the peak twitch force of the knee extensors evoked by electrical stimulation at 10 Hz (Db10). It is likely that reductions in maximal voluntary contraction may be related to an excitation-contraction coupling failure (i.e. low-frequency fatigue) as suggest the reduction in the Db10. The current results confirm that acute changes in jump performance may not be appropriate to evaluate acute fatigue in endurance trained runners.

Utilizing the RPE-Clamp model to examine interactions among factors associated with perceived fatigability and performance fatigability in women and men.

The purpose of the present study was to examine the interactions between perceived fatigability and performance fatigability in women and men by utilizing the RPE-Clamp model to assess the fatigue-induced effects of a sustained, isometric forearm flexion task anchored to RPE = 8 on time to task failure (TTF), torque, and neuromuscular responses. Twenty adults (10 men and 10 women) performed two, 3s forearm flexion maximal voluntary isometric contractions (MVICs) followed by a sustained, isometric forearm flexion task anchored to RPE = 8 using the OMNI-RES (0-10) scale at an elbow joint angle of 100°. Electromyographic amplitude (EMG AMP) was recorded from the biceps brachii. Torque and EMG AMP values resulting from the sustained task were normalized to the pretest MVIC. Neuromuscular efficiency was defined as NME = normalized torque/normalized EMG AMP. Mixed factorial ANOVAs and Bonferroni corrected dependent t tests and independent t tests were used to examine differences across time and between sex for torque and neuromuscular parameters. There were no differences between the women and men for the fatigue-induced decreases in torque, EMG AMP, or NME, and the mean decreases (collapsed across sex) were 50.3 ± 8.6 to 2.8 ± 2.9% MVIC, 54.7 ± 12.0 to 19.6 ± 5.3% MVIC, and 0.94 ± 0.19 to 0.34 ± 0.16, respectively. Furthermore, there were no differences between the women and men for TTF (251.8 ± 74.1vs.258.7 ± 77.9s). The results suggested that the voluntary reductions in torque to maintain RPE and the decreases in NME were likely due to group III/IV afferent feedback from peripheral fatigue that resulted in excitation-contraction coupling failure.

The role of action potential changes in depolarization-induced failure of excitation contraction coupling in mouse skeletal muscle.

Excitation-contraction coupling (ECC) is the process by which electrical excitation of muscle is converted into force generation. Depolarization of skeletal muscle resting potential contributes to failure of ECC in diseases such as periodic paralysis, intensive care unit acquired weakness and possibly fatigue of muscle during vigorous exercise. When extracellular K+ is raised to depolarize the resting potential, failure of ECC occurs suddenly, over a narrow range of resting potentials. Simultaneous imaging of Ca2+ transients and recording of action potentials (APs) demonstrated failure to generate Ca2+ transients when APs peaked at potentials more negative than -30mV. An AP property that closely correlated with failure of the Ca2+ transient was the integral of AP voltage with respect to time. Simultaneous recording of Ca2+ transients and APs with electrodes separated by 1.6mm revealed AP conduction fails when APs peak below -21mV. We hypothesize propagation of APs and generation of Ca2+ transients are governed by distinct AP properties: AP conduction is governed by AP peak, whereas Ca2+ release from the sarcoplasmic reticulum is governed by AP integral. The reason distinct AP properties may govern distinct steps of ECC is the kinetics of the ion channels involved. Na channels, which govern propagation, have rapid kinetics and are insensitive to AP width (and thus AP integral) whereas Ca2+ release is governed by gating charge movement of Cav1.1 channels, which have slower kinetics such that Ca2+ release is sensitive to AP integral. The quantitative relationships established between resting potential, AP properties, AP conduction and Ca2+ transients provide the foundation for future studies of failure of ECC induced by depolarization of the resting potential.

Effects of fatigue and recovery on electromechanical delay during isokinetic muscle actions

Objective: To examine muscle-specific differences and the effects of fatigue and recovery on electromechanical delay (EMD) during maximal isokinetic muscle actions. Approach: Thirteen men performed maximal isokinetic knee extension muscle actions at 60° s−1, pretest, posttest, and after 5 min of recovery from 25 maximal isokinetic knee extensions. The onsets of the electromyographic, mechanomyographic, and force signals were used to identify EMD measures from the vastus lateralis (VL), vastus medialis (VM), and rectus femoris (RF). Main results: There were posttest increases in all EMD measures for all muscles that returned to pretest levels after 5 min of recovery. There were, however, no differences in EMD measures between the VL and VM. All EMD values from the RF were greater than the VL and VM. Significance: These findings suggested muscle-specific differences in EMD and that excitation-contraction coupling failure and increased compliance of the series elastic component occurred posttest, but subsided after 5 min of recovery.

Effects of Fatigue on Voluntary Electromechanical and Relaxation Electromechanical Delay.

The purposes of the present study were to examine: 1) the effects of fatigue on electromechanical delay from the onsets of the electromyographic signal to force production (EMDE-F), the onsets of the electromyographic to mechanomyographic signals (EMDE-M), the onsets of the mechanomyographic signal to force production (EMDM-F), as well as the cessations of the electromyographic to force production (R-EMDE-F), cessation of the electromyographic to mechanomyographic signals (R-EMDE-M), and cessations of the mechanomyographic signal to force production (R-EMDM-F); and 2) the relative contributions from EMDE-M and EMDM-F to EMDE-F as well as R-EMDE-M and R-EMDM-F to R-EMDE-F from the vastus lateralis in non-fatigued and fatigued states. The values EMDE-F, EMDE-M, EMDM-F, R-EMDE-F, R-EMDE-M and R-EMDM-F were calculated during maximal voluntary isometric contractions, before and after 70% 1-repetition maximum leg extensions to failure. There were significant pretest to posttest increases in EMDE-F (73%;p<0.01), EMDE-M (99%;p<0.01), EMDM-F (60%;p<0.01), R-EMDE-F (101%;p<0.01) and R-EMDM-F (368%;p<0.01), but no significant change in R-EMDE-M (25%;p=0.46). Fatigue-induced increase in EMDE-F indicated excitation-contraction coupling failure (EMDE-M) and increases in the compliance of the series elastic component (EMDM-F). Increases in R-EMDE-F were due to increases in relaxation time for the series elastic component (R-EMDM-F), but not changes in the reversal of excitation-contraction coupling (R-EMDE-M).

Changes in electromechanical delay during fatiguing dynamic muscle actions.

The onsets of the electromyographic (EMG) signal, mechanomyographic (MMG) signal, and force production were used to identify voluntary electromechanical delay (EMD) during maximal and submaximal leg extensions. Twelve men performed 30% 1-repetition-maximum concentric, dynamic constant external resistance leg extensions to failure. The time periods for the onsets of EMG to MMG (EMDE-M ), MMG to force (EMDM-F ), and EMG to force (EMDE-F ) were assessed before, during, and after testing. Pretest vs. posttest assessments indicated approximately equal contributions from excitation-contraction coupling (EMDE-M ) and series elastic component (EMDM-F ) to the overall EMDE-F . During the workbout, excitation-contraction coupling failure and increases in the compliance of the series elastic component began at 40% and 30% of the repetitions to failure, respectively, and continued to failure. Increases in the series elastic component (EMDM-F ) began to affect EMDE-F before excitation-contraction coupling failure (EMDE-M ), and both contributed to EMDE-F throughout the remainder of the fatiguing workbout. Muscle Nerve 56: 315-320, 2017.

The relationship between oxygen uptake kinetics and neuromuscular fatigue in high-intensity cycling exercise.

In theory, a slow oxygen uptake ([Formula: see text]) kinetics leads to a greater accumulation of anaerobic by-products, which can, in turn, induce more neuromuscular fatigue. However, the existence of this relationship has never been tested. After two sessions to measure peak [Formula: see text], peak power output (POpeak), and [Formula: see text] kinetics responses in the unfatigued state (τ [Formula: see text] MOD), 10 healthy young adults performed a 6-min cycling bout at 80% POpeak (INT6-min). [Formula: see text] kinetics responses were also measured during INT6-min. Neuromuscular fatigue was measured isometrically pre- and post-INT6-min (immediately post- and 15-s post-INT6-min) with an innovative cycle ergometer. Maximal voluntary contraction (MVC) force, high-frequency doublet amplitude, and the ratio of low- to high-frequency doublet amplitudes decreased by 34 ± 7, 43 ± 11, and 31 ± 13%, respectively (all P < 0.01). A significant Spearman's rank correlation was observed between the change in low-frequency doublet force (ΔDb10) immediately after INT6-min and both τ [Formula: see text] MOD and τ [Formula: see text] INT6-min (ρ = -0.68 and ρ = -0.67, both P < 0.05). When considering the largest responses from the two neuromuscular evaluations post-INT6-min, significant correlations were also found between τ [Formula: see text] MOD and ΔDb10 (ρ = -0.74; P < 0.05) and between τ[Formula: see text] INT6-min and both ΔDb10 and low-frequency fatigue (ρ = -0.70 and ρ = -0.66; both P < 0.05). The present results suggest that subjects with slow [Formula: see text] kinetics experience more peripheral fatigue, in particular more excitation-contraction coupling failure, likely due to a greater accumulation of protons and/or inorganic phosphates.

Protein Fragmentation As a Regulatory Mechanism: Insights from Two Different Ca2+ Channels, RyR1 and IP3R.

The ryanodine receptor 1 (RyR1) is the major skeletal muscle Ca2+ release channel and as such a key player in excitation-contraction coupling. Intriguingly, a recent paper reported RyR1 fragmentation 24 h after high-intensity interval exercise (six 30-s all-out cycling bouts with 4 min recoveries in between) in vastus lateralis muscle biopsies taken from recreationally active men (Place et al., 2015). In contrast to what could have been hypothesized based on the role of RyR1 in excitation-contraction coupling, this RyR1 fragmentation did not appear to result in excitation-contraction coupling failure, as both maximal voluntary contraction force and forces evoked by supramaximal electrical stimulations had fully recovered 24 h after exercise. Overall, these results suggest that fragmented RyR1 retain their capacity to release Ca2+ in response to an action potential.

Fatigue associated with prolonged graded running.

Scientific experiments on running mainly consider level running. However, the magnitude and etiology of fatigue depend on the exercise under consideration, particularly the predominant type of contraction, which differs between level, uphill, and downhill running. The purpose of this review is to comprehensively summarize the neurophysiological and biomechanical changes due to fatigue in graded running. When comparing prolonged hilly running (i.e., a combination of uphill and downhill running) to level running, it is found that (1) the general shape of the neuromuscular fatigue-exercise duration curve as well as the etiology of fatigue in knee extensor and plantar flexor muscles are similar and (2) the biomechanical consequences are also relatively comparable, suggesting that duration rather than elevation changes affects neuromuscular function and running patterns. However, 'pure' uphill or downhill running has several fatigue-related intrinsic features compared with the level running. Downhill running induces severe lower limb tissue damage, indirectly evidenced by massive increases in plasma creatine kinase/myoglobin concentration or inflammatory markers. In addition, low-frequency fatigue (i.e., excitation-contraction coupling failure) is systematically observed after downhill running, although it has also been found in high-intensity uphill running for different reasons. Indeed, low-frequency fatigue in downhill running is attributed to mechanical stress at the interface sarcoplasmic reticulum/T-tubule, while the inorganic phosphate accumulation probably plays a central role in intense uphill running. Other fatigue-related specificities of graded running such as strategies to minimize the deleterious effects of downhill running on muscle function, the difference of energy cost versus heat storage or muscle activity changes in downhill, level, and uphill running are also discussed.

The role of proteases in excitation-contraction coupling failure in muscular dystrophy.

Duchenne muscular dystrophy (DMD) is one of the most frequent types of muscular dystrophy. Alterations in intracellular calcium (Ca(2+)) handling are thought to contribute to the disease severity in DMD, possibly due to the activation of Ca(2+)-activated proteases. The purpose of this study was twofold: 1) to determine whether prolonged excitation-contraction (E-C) coupling disruption following repeated contractions is greater in animals lacking both dystrophin and utrophin (mdx/Utr(-/-)) compared with mice lacking only dystrophin (mdx); and 2) to assess whether protease inhibition can prevent E-C coupling failure following repeated tetani in these dystrophic mouse models. Excitation-contraction coupling was assessed using Fura-2 ratio, as an index of intracellular free Ca(2+) concentration, in response to electrical stimulation of single muscle fibers from the flexor digitorum brevis muscle. Resting Fura-2 ratio was higher in dystrophic compared with control (Con) fibers, but peak Fura-2 ratios during stimulation were similar in dystrophic and Con fibers. One hour after a series of repeated tetani, peak Fura-2 ratios were reduced by 30 ± 5.6%, 23 ± 2%, and 36 ± 3.1% in mdx, mdx/Utr(+/-), and mdx/Utr(-/-), respectively, with the greatest reduction in mdx/Utr(-/-) fibers (P < 0.05). Protease inhibition attenuated this decrease in peak Fura-2 ratio. These data indicate that E-C coupling impairment after repeated contractions is greatest in fibers lacking both dystrophin and utrophin and that prevention of protease activation can mitigate the prolonged E-C coupling impairment. These data further suggest that acute protease inhibition may be useful in reducing muscle weakness in DMD.

Chemical Uncoupling the DHPR-RyR1 Complex by Substituted Halogenated Biphenyls and Diphenylethers

Ca2+ release units (CRU) of skeletal muscle have two components necessary for excitation-contraction coupling (ECC); the DHPR and RyR1 that physically interact to engage conformational coupling that involves both orthograde and retrograde regulation of Ca2+ dynamics during ECC and SR Ca2+ leak when the muscle is at rest. We previously identified that 2,2',3,5',6-pentachlorobiphenyl (PCB95) promotes long-lived stability of the full open state of RyR1 channels (Samso et al 2009). Further investigations PCB95 and its 4- and 5-hydroxy (-OH) derivatives toward rabbit skeletal muscle ryanodine receptor (RyR1) were performed using [3H]ryanodine binding and SR Ca2+ flux analyses. 5-OH metabolites have comparable activity to their respective parent in both assays; however, 4-OH derivatives are unable to trigger Ca2+ release from SR microsomes in the presence of Ca2+-ATPase activity. PCB95 and derivatives are investigated using single channel voltage-clamp and primary murine embryonic muscle cells. Like PCB95, 5-OH-PCB95 quickly and persistently increases channel open probability (Po>0.9) by stabilizing the full-open channel state, whereas 4-OH-PCB95 transiently enhances Po. Ca2+ imaging of myotubes show acute exposure to PCB95 (5μM) potentiates ECC and caffeine responses and partially depletes SR Ca2+ stores. Exposure to 5-OH-PCB95 (5μM) increases cytoplasmic Ca2+, leading to ECC failure in 50% of myotubes with the remainder retaining negligible responses. 4-OH-PCB95 neither increases baseline Ca2+ nor causes ECC failure but depresses ECC and caffeine responses by 50%. With longer (3 h) exposure to 300nM PCB95, 5-OH-PCB95, or 4-OH-PCB95 decreases the number of ECC responsive myotubes by 22%, 81%, and 51% compared with control by depleting SR Ca2+ and/or uncoupling ECC. The position of hydroxylation of PCB95 and chemically related diphenylethers differentially influence RyR1 channel gating kinetics, SR Ca2+ leak, and the pattern of ECC impairment, including ECC uncoupling in intact muscle cells. P01 AR52354.

Structure-Activity Relationship of Selected Meta- and Para-Hydroxylated Non–Dioxin Like Polychlorinated Biphenyls: From Single RyR1 Channels to Muscle Dysfunction

Non-dioxin like polychlorinated biphenyls (NDL-PCBs) are legacy environmental contaminants with contemporary unintentional sources. NDL-PCBs interact with ryanodine receptors (RyRs), Ca(2+) channels of sarcoplasmic/endoplasmic reticulum (SR/ER) that regulate excitation-contraction coupling (ECC) and Ca(2+)-dependent cell signaling in muscle. Activities of 4 chiral congeners PCB91, 95, 132, and 149 and their respective 4- and 5-hydroxy (-OH) derivatives toward rabbit skeletal muscle ryanodine receptor (RyR1) are investigated using [(3)H]ryanodine binding and SR Ca(2+) flux analyses. Although 5-OH metabolites have comparable activity to their respective parent in both assays, 4-OH derivatives are unable to trigger Ca(2+) release from SR microsomes in the presence of Ca(2+)-ATPase activity. PCB95 and derivatives are investigated using single channel voltage-clamp and primary murine embryonic muscle cells (myotubes). Like PCB95, 5-OH-PCB95 quickly and persistently increases channel open probability (p o > .9) by stabilizing the full-open channel state, whereas 4-OH-PCB95 transiently enhances p o. Ca(2+) imaging of myotubes loaded with Fluo-4 show that acute exposure to PCB95 (5 µM) potentiates ECC and caffeine responses and partially depletes SR Ca(2+) stores. Exposure to 5-OH-PCB95 (5 µM) increases cytoplasmic Ca(2+), leading to rapid ECC failure in 50% of myotubes with the remainder retaining negligible responses. 4-OH-PCB95 neither increases baseline Ca(2+) nor causes ECC failure but depresses ECC and caffeine responses by 50%. With longer (3h) exposure to 300 nM PCB95, 5-OH-PCB95, or 4-OH-PCB95 decreases the number of ECC responsive myotubes by 22%, 81%, and 51% compared with control by depleting SR Ca(2+) and/or uncoupling ECC. NDL-PCBs and their 5-OH and 4-OH metabolites differentially influence RyR1 channel activity and ECC in embryonic skeletal muscle.

Ca2+‐dependent proteolysis of junctophilin‐1 and junctophilin‐2 in skeletal and cardiac muscle

Excessive increases in intracellular [Ca(2+)] in skeletal muscle fibres cause failure of excitation-contraction coupling by disrupting communication between the dihydropyridine receptors in the transverse tubular system and the Ca(2+) release channels (RyRs) in the sarcoplasmic reticulum (SR), but the exact mechanism is unknown. Previous work suggested a possible role of Ca(2+)-dependent proteolysis in this uncoupling process but found no proteolysis of the dihydropyridine receptors, RyRs or triadin. Junctophilin-1 (JP1; ∼90 kDa) stabilizes close apposition of the transverse tubular system and SR membranes in adult skeletal muscle; its C-terminal end is embedded in the SR and its N-terminal associates with the transverse tubular system membrane. Exposure of skeletal muscle homogenates to precisely set [Ca(2+)] revealed that JP1 undergoes Ca(2+)-dependent proteolysis over the physiological [Ca(2+)] range in tandem with autolytic activation of endogenous μ-calpain. Cleavage of JP1 occurs close to the C-terminal, yielding a ∼75 kDa diffusible fragment and a fixed ∼15 kDa fragment. Depolarization-induced force responses in rat skinned fibres were abolished following 1 min exposure to 40 μm Ca(2+), with accompanying loss of full-length JP1. Supraphysiological stimulation of rat skeletal muscle in vitro by repeated tetanic stimulation in 30 mm caffeine also produced marked proteolysis of JP1 (and not RyR1). In dystrophic mdx mice, JP1 proteolysis is seen in limb muscles at 4 and not at 10 weeks of age. Junctophilin-2 in cardiac and skeletal muscle also undergoes Ca(2+)-dependent proteolysis, and junctophilin-2 levels are reduced following cardiac ischaemia-reperfusion. Junctophilin proteolysis may contribute to skeletal muscle weakness and cardiac dysfunction in a range of circumstances.

Effects of in vivo injury on the neuromuscular junction in healthy and dystrophic muscles

The most common and severe form of muscular dystrophy is Duchenne muscular dystrophy (DMD), a disorder caused by the absence of dystrophin, a structural protein found on the cytoplasmic surface of the sarcolemma of striated muscle fibres. Considerable attention has been dedicated to studying myofibre damage and muscle plasticity, but there is little information to determine if damage from contraction-induced injury occurs at or near the nerve terminal axon. We used α-bungarotoxin to compare neuromuscular junction (NMJ) morphology in healthy (wild-type, WT) and dystrophic (mdx) mouse quadriceps muscles and evaluated transcript levels of the post-synaptic muscle-specific kinase signalling complex. Our focus was to study changes in NMJs after injury induced with an established in vivo animal injury model. Neuromuscular transmission, electromyography (EMG), and NMJ morphology were assessed 24 h after injury. In non-injured muscle, muscle-specific kinase expression was significantly decreased in mdx compared to WT. Injury resulted in a significant loss of maximal torque in WT (39 ± 6%) and mdx (76 ± 8%) quadriceps, but significant changes in NMJ morphology, neuromuscular transmission and EMG data were found only in mdx following injury. Compared with WT mice, motor end-plates of mdx mice demonstrated less continuous morphology, more disperse acetylcholine receptor aggregates and increased number of individual acetylcholine receptor clusters, an effect that was exacerbated following injury. Neuromuscular transmission failure increased and the EMG measures decreased after injury in mdx mice only. The data show that eccentric contraction-induced injury causes morphological and functional changes to the NMJs in mdx skeletal muscle, which may play a role in excitation-contraction coupling failure and progression of the dystrophic process.

Triclosan impairs excitation–contraction coupling and Ca 2+ dynamics in striated muscle

Triclosan (TCS), a high-production-volume chemical used as a bactericide in personal care products, is a priority pollutant of growing concern to human and environmental health. TCS is capable of altering the activity of type 1 ryanodine receptor (RyR1), but its potential to influence physiological excitation-contraction coupling (ECC) and muscle function has not been investigated. Here, we report that TCS impairs ECC of both cardiac and skeletal muscle in vitro and in vivo. TCS acutely depresses hemodynamics and grip strength in mice at doses ≥12.5 mg/kg i.p., and a concentration ≥0.52 μM in water compromises swimming performance in larval fathead minnow. In isolated ventricular cardiomyocytes, skeletal myotubes, and adult flexor digitorum brevis fibers TCS depresses electrically evoked ECC within ∼10-20 min. In myotubes, nanomolar to low micromolar TCS initially potentiates electrically evoked Ca(2+) transients followed by complete failure of ECC, independent of Ca(2+) store depletion or block of RyR1 channels. TCS also completely blocks excitation-coupled Ca(2+) entry. Voltage clamp experiments showed that TCS partially inhibits L-type Ca(2+) currents of cardiac and skeletal muscle, and [(3)H]PN200 binding to skeletal membranes is noncompetitively inhibited by TCS in the same concentration range that enhances [(3)H]ryanodine binding. TCS potently impairs orthograde and retrograde signaling between L-type Ca(2+) and RyR channels in skeletal muscle, and L-type Ca(2+) entry in cardiac muscle, revealing a mechanism by which TCS weakens cardiac and skeletal muscle contractility in a manner that may negatively impact muscle health, especially in susceptible populations.

Neuromuscular Consequences of an Extreme Mountain Ultra-Marathon

We investigated the physiological consequences of one of the most extreme exercises realized by humans in race conditions: a 166-km mountain ultra-marathon (MUM) with 9500 m of positive and negative elevation change. For this purpose, (i) the fatigue induced by the MUM and (ii) the recovery processes over two weeks were assessed. Evaluation of neuromuscular function (NMF) and blood markers of muscle damage and inflammation were performed before and immediately following (n = 22), and 2, 5, 9 and 16 days after the MUM (n = 11) in experienced ultra-marathon runners. Large maximal voluntary contraction decreases occurred after MUM (−35% [95% CI: −28 to −42%] and −39% [95% CI: −32 to −46%] for KE and PF, respectively), with alteration of maximal voluntary activation, mainly for KE (−19% [95% CI: −7 to −32%]). Significant modifications in markers of muscle damage and inflammation were observed after the MUM as suggested by the large changes in creatine kinase (from 144±94 to 13,633±12,626 UI L−1), myoglobin (from 32±22 to 1,432±1,209 µg L−1), and C-Reactive Protein (from <2.0 to 37.7±26.5 mg L−1). Moderate to large reductions in maximal compound muscle action potential amplitude, high-frequency doublet force, and low frequency fatigue (index of excitation-contraction coupling alteration) were also observed for both muscle groups. Sixteen days after MUM, NMF had returned to initial values, with most of the recovery process occurring within 9 days of the race. These findings suggest that the large alterations in NMF after an ultra-marathon race are multi-factorial, including failure of excitation-contraction coupling, which has never been described after prolonged running. It is also concluded that as early as two weeks after such an extreme running exercise, maximal force capacities have returned to baseline.

Alterations of neuromuscular function after prolonged running, cycling and skiing exercises.

It is well known that impairment of performance resulting from muscle fatigue differs according to the types of contraction involved, the muscular groups tested and the exercise duration/intensity. Depending on these variables, strength loss with fatigue can originate from several sites from the motor cortex through to contractile elements. This has been termed 'task dependency of muscle fatigue'. Only recently have studies focused on the origin of muscle fatigue after prolonged exercise lasting 30 minutes to several hours. Central fatigue has been shown to contribute to muscle fatigue during long-distance running by using different methods such as the twitch interpolation technique, the ratio of the electromyogram (EMG) signal during maximal voluntary contraction normalised to the M-wave amplitude or the comparison of the forces achieved with voluntary- and electrically-evoked contractions. Some central activation deficit has also been observed for knee extensor muscles in cycling but central fatigue after activities inducing low muscular damage was attenuated compared with running. While supraspinal fatigue cannot be ruled out, it can be suggested that spinal adaptation, such as inhibition from type III and IV group afferents or disfacilitation from muscle spindles, contributes to the reduced neural drive after prolonged exercise. It has been shown that after a 30 km run, individuals with the greatest knee extensor muscle strength loss experienced a significant activation deficit. However, central fatigue alone cannot explain the entire strength loss after prolonged exercise. Alterations of neuromuscular propagation, excitation-contraction coupling failure and modifications of the intrinsic capability of force production may also be involved. Electrically-evoked contractions and associated EMG can help to characterise peripheral fatigue. The purpose of this review is to further examine the central and peripheral mechanisms contributing to strength loss after prolonged running, cycling and skiing exercises.

LOW-FREQUENCY FATIGUE INCREASES WITH THE NUMBER OF STRETCHES OF ACTIVATED RAT SKELETAL MUSCLES

PURPOSE To examine the relationship between stretch number of activated plantar flexor muscles and low-frequency fatigue. METHODS In female Sprague Dawley rats (age 3–5 mo), nerve stimulation was used to produce muscle contractions. Stretches [3 stretches (S), 10S, 60S or 240S (n = 6 in each group)] were imposed on isometric contractions [80 Hz, 0.2 ms pulse duration, 5.3±0.3 V (mean±SE)] by ankle rotation from 90° to 40° (velocity: 600 °·s−1, simulation time: 0.9 s, inter-stretch rest time: 40 s). Forty seconds after the last stretch, an isometric contraction (80 Hz at 90°) was performed. Force-frequency relationships with isometric contractions at 90° were measured before and 1 hour after the stretch protocols. An index for low-frequency fatigue was calculated by the change in the 20:100 Hz isometric force ratio. RESULTS All stretch protocols resulted in isometric force deficits (3S: 37.0±2.4%, 10S: 44.6±1.5%, 60S: 74.1±1.9%, 240S: 83.4±2.0%). After 1 hour, substantial deficits remained (3S: 33.3±1.7%, 10S: 37.2±2.3%, 60S: 67.6±1.5%, 240S: 77.7±1.2%) and low-frequency fatigue was present in all groups. Force deficits with contractions at 20 Hz were significantly larger than at 100 Hz (3S: 45.9±2.4% vs. 30.8±2.0%, 10S: 57.1±3.0% vs. 34.2±2.1%, 60S: 78.9±0.4% vs. 63.0±1.7%, 240S: 88.5±1.3% vs. 73.9±1.4%). Low-frequency fatigue became significantly larger with an increase in the number of stretches (P = 0.0009). CONCLUSIONS The amount of low-frequency fatigue was related to the number of stretches of activated skeletal muscles providing indirect evidence for an increased failure of excitation-contraction coupling with stretch number. Supported by NIOSH R01-OH-02918.

Heart failure after myocardial infarction: altered excitation-contraction coupling.

Heart failure (HF) frequently follows the occurrence of myocardial infarction (MI). Questions about how HF develops and what cellular defects contribute to this dysfunction led to this study. Methods and Results-- MI was induced in rats by coronary artery ligation. Clinical examination of the post-MI (PMI) surviving animals indicated that they were in overt HF by all measures. Cellular examination of the cardiomyocytes by patch-clamp and confocal [Ca(2+)](i) imaging methods indicated that cellular function was significantly compromised. At the single-cell level, [Ca(2+)](i) transient amplitudes were reduced and contractions were decreased and slowed, although Ca(2+) current (I(Ca)) remained unchanged. The excitation-contraction coupling (ECC) gain function measured as Delta[Ca(2+)](i)/I(Ca) was significantly decreased. Ouabain, a cardiotonic steroid that blocks the Na(+),K(+)-ATPase and activates Ca(2+) entry via cardiac Na(+) channels, largely alleviated this defect. After MI, I(Ca) becomes less able to trigger release of Ca(2+) from the sarcoplasmic reticulum. This failure of ECC is a major factor contributing to the development of contractile dysfunction and HF in PMI animals. The improved ECC gain, enhanced Ca(2+) entry, and augmented Ca(2+) signaling due to cardiotonic steroids contribute to the beneficial effects of these agents.