Nicolas Place

ASSESSMENT OF THE RELIABILITY OF CENTRAL AND PERIPHERAL FATIGUE AFTER SUSTAINED MAXIMAL VOLUNTARY CONTRACTION OF THE QUADRICEPS MUSCLE

The aim of the present study was to further confirm the validity of measurements for characterizing neuromuscular alterations by establishing their reliability both before and after fatigue. Thirteen men (28 ± 5 years) volunteered to participate in two separate identical sessions requiring the performance of a sustained maximal voluntary contraction (MVC) with the quadriceps muscle for 2 min. MVC and transcutaneous electrical stimulations were used before and immediately after the fatiguing contraction to investigate the reliability of MVC torque, central activation, and peripheral variables (M‐wave properties, peak twitch, peak doublet) within and between sessions. Based on previous and present results, we advise the use of (1) voluntary activation level with potentiated doublet as a reference to describe central fatigue, (2) electromyographic activity of vastus lateralis muscle as a surrogate for quadriceps for both voluntary and evoked contraction, and (3) potentiated peak doublet amplitude to investigate contractile fatigue. These findings can be useful in the choice of the parameters describing central and peripheral fatigue of the quadriceps muscle in future studies. Muscle Nerve, 2007

Time Course of Neuromuscular Alterations during a Prolonged Running Exercise

PURPOSE This study investigated the time course of contractile and neural alterations of knee extensor (KE) muscles during a long-duration running exercise. METHODS Nine well-trained triathletes and endurance runners sustained 55% of their maximal aerobic velocity (MAV) on a motorized treadmill for a period of 5 h. Maximal voluntary contraction (MVC), maximal voluntary activation level (%VA), and electrically evoked contractions (single and tetanic stimulations) of KE muscles were evaluated before, after each hour of exercise during short (10 min) interruptions, and at the end of the 5-h period. Oxygen uptake was also measured at regular intervals during the exercise. RESULTS Reductions of MVC and %VA were significant after the 4th hour of exercise and reached -28% (P < 0.001) and -16% (P < 0.01) respectively at the end of the exercise. The reduction in MVC was highly correlated with the decline of %VA (r = 0.98, P < 0.001). M-wave was also altered after the fourth hour of exercise (P < 0.05) in both vastus lateralis and rectus femoris muscles. Peak twitch was potentiated at the end of the exercise (+18%, P = 0.01); 20- and 80-Hz maximal tetanic forces were not altered by the exercise. Oxygen uptake increased linearly during the running period (+18% at 5 h, P < 0.001). CONCLUSION These findings suggest that KE maximal voluntary force generating capability is depressed in the final stages of a 5-h running exercise. Central activation failure and alterations in muscle action potential transmission were important mechanisms contributing to the impairment of the neuromuscular function during prolonged running.

Muscle fatigue: from observations in humans to underlying mechanisms studied in intact single muscle fibres

Prolonged dynamic exercise and sustained isometric contractions induce muscle fatigue, as manifested by decreased performance and a reduction in the maximum voluntary contraction force. Studies with non-invasive measurements in exercising humans show that mechanisms located beyond the sarcolemma are important in the fatigue process. In this review, we describe probable cellular mechanisms underlying fatigue-induced changes in excitation–contraction (E–C) coupling occurring in human muscle fibres during strenuous exercise. We use fatigue-induced changes observed in intact single muscle fibres, where force and cellular Ca2+ handling can be directly measured, to explain changes in E–C coupling observed in human muscle during exercise.

Reactive oxygen species and fatigue-induced prolonged low-frequency force depression in skeletal muscle fibres of rats, mice and SOD2 overexpressing mice

Skeletal muscle often shows a delayed force recovery after fatiguing stimulation, especially at low stimulation frequencies. In this study we focus on the role of reactive oxygen species (ROS) in this fatigue‐induced prolonged low‐frequency force depression. Intact, single muscle fibres were dissected from flexor digitorum brevis (FDB) muscles of rats and wild‐type and superoxide dismutase 2 (SOD2) overexpressing mice. Force and myoplasmic free [Ca2+] ([Ca2+]i) were measured. Fibres were stimulated at different frequencies before and 30 min after fatigue induced by repeated tetani. The results show a marked force decrease at low stimulation frequencies 30 min after fatiguing stimulation in all fibres. This decrease was associated with reduced tetanic [Ca2+]i in wild‐type mouse fibres, whereas rat fibres and mouse SOD2 overexpressing fibres instead displayed a decreased myofibrillar Ca2+ sensitivity. The SOD activity was ∼50% lower in wild‐type mouse than in rat FDB muscles. Myoplasmic ROS increased during repeated tetanic stimulation in rat fibres but not in wild‐type mouse fibres. The decreased Ca2+ sensitivity in rat fibres could be partially reversed by application of the reducing agent dithiothreitol, whereas the decrease in tetanic [Ca2+]i in wild‐type mouse fibres was not affected by dithiothreitol or the antioxidant N‐acetylcysteine. In conclusion, we describe two different causes of fatigue‐induced prolonged low‐frequency force depression, which correlate to differences in SOD activity and ROS metabolism. These findings may have clinical implications since ROS‐mediated impairments in myofibrillar function can be counteracted by reductants and antioxidants, whereas changes in SR Ca2+ handling appear more resistant to interventions.

Alterations of Neuromuscular Function after the World's Most Challenging Mountain Ultra-Marathon

We investigated the physiological consequences of the most challenging mountain ultra-marathon (MUM) in the world: a 330-km trail run with 24000 m of positive and negative elevation change. Neuromuscular fatigue (NMF) was assessed before (Pre-), during (Mid-) and after (Post-) the MUM in experienced ultra-marathon runners (n = 15; finish time  = 122.43 hours ±17.21 hours) and in Pre- and Post- in a control group with a similar level of sleep deprivation (n = 8). Blood markers of muscle inflammation and damage were analyzed at Pre- and Post-. Mean ± SD maximal voluntary contraction force declined significantly at Mid- (−13±17% and −10±16%, P<0.05 for knee extensor, KE, and plantar flexor muscles, PF, respectively), and further decreased at Post- (−24±13% and −26±19%, P<0.01) with alteration of the central activation ratio (−24±24% and −28±34% between Pre- and Post-, P<0.05) in runners whereas these parameters did not change in the control group. Peripheral NMF markers such as 100 Hz doublet (KE: −18±18% and PF: −20±15%, P<0.01) and peak twitch (KE: −33±12%, P<0.001 and PF: −19±14%, P<0.01) were also altered in runners but not in controls. Post-MUM blood concentrations of creatine kinase (3719±3045 Ul·1), lactate dehydrogenase (1145±511 UI·L−1), C-Reactive Protein (13.1±7.5 mg·L−1) and myoglobin (449.3±338.2 µg·L−1) were higher (P<0.001) than at Pre- in runners but not in controls. Our findings revealed less neuromuscular fatigue, muscle damage and inflammation than in shorter MUMs. In conclusion, paradoxically, such extreme exercise seems to induce a relative muscle preservation process due likely to a protective anticipatory pacing strategy during the first half of MUM and sleep deprivation in the second half.

Mechanisms of fatigue induced by isometric contractions in exercising humans and in mouse isolated single muscle fibres

1 Muscle fatigue (i.e. the decrease in muscle performance during exercise) has been studied extensively using a variety of experimental paradigms, from mouse to human, from single cell to whole‐body exercise. Given the disparity of models used to characterize muscle fatigue, it can be difficult to establish whether the results of basic in vitro studies are applicable to exercise in humans. 2 In the present brief review, our attempt is to relate neuromuscular alterations caused by repeated or sustained isometric contraction in humans to changes in excitation–contraction (E‐C) coupling observed in intact single muscle fibres, where force and the free myoplasmic [Ca2+] can be measured. 3 Accumulated data indicate that impairment of E‐C coupling, most likely located within muscle fibres, accounts for the fatigue‐induced decrease in maximal force in humans, whereas central (neural) fatigue is of greater importance for the inability to continue a sustained low‐intensity contraction. Based on data from intact single muscle fibres, the fatigue‐induced impairment in E‐C coupling involves: (i) a reduced number of active cross‐bridges owing to a decreased release of Ca2+; (ii) a decreased sensitivity of the myofilaments to Ca2+; and/or (iii) a reduced force produced by each active cross‐bridge. 4 In conclusion, data from single muscle fibre studies can be used to increase our understanding of fatigue mechanisms in some, but not all, types of human exercise. To further increase the understanding of fatigue mechanisms in humans, we propose future studies using in vitro stimulation patterns that are closer to the in vivo situation.

Comparison of quadriceps inactivation between nerve and muscle stimulation

We evaluated the use of direct muscle stimulation for quantifying quadriceps inactivation at different contraction levels as opposed to conventional twitch interpolation using nerve stimulation. Fourteen healthy volunteers were tested. Paired stimuli were delivered to the femoral nerve or to the quadriceps muscle belly during voluntary contractions ranging from 20% to 100% of maximum, and the amplitude of the superimposed doublet was quantified to investigate inactivation. Superimposed doublet for muscle and nerve stimulation, respectively between the range of 60% to 100% of maximum (e.g., at 100%, muscle stimulation was 14 ± 5 Nm and nerve stimulation was 15 ± 6 Nm). Despite higher current doses, muscle stimulation was associated with less discomfort than nerve stimulation (P < 0.05). Collectively, our data suggest that direct muscle stimulation could be used to assess quadriceps inactivation at maximal and quasi‐maximal contraction levels as a valid alternative to motor nerve stimulation. Muscle Nerve, 2010

Nonshivering thermogenesis protects against defective calcium handling in muscle

When acutely exposed to a cold environment, mammals shiver to generate heat. During prolonged cold exposure, shivering is replaced by adaptive adrenergic nonshivering thermogenesis with increased heat production in brown adipose tissue due to activation of uncoupling protein‐1 (UGP1). This cold acclimation is associated with chronically increased sympathetic stimulation of skeletal muscle, which may increase the sarcoplasmic reticulum (SR) Ca2+ leak via destabilized ryanodine receptor 1 (RyR1) channel complexes. Here, we use genetically engineered UGP1‐deficient (UGP1‐KO) mice that rely completely on shivering in the cold. We examine soleus muscle, which participates in shivering, and flexor digitorum brevis (FDB) muscle, a distal and superficial muscle that does not shiver. Soleus muscles of cold‐acclimated UGP1‐KO mice exhibited severe RyR1 PKA hyperphosphorylation and calstabin1 depletion, as well as markedly decreased SR Ca2+ release and force during contractions. In stark contrast, the RyR1 channel complexes were little affected, and Ca2+ and force were not decreased in FDB muscles of cold‐acclimated UGP1‐KO mice. These results indicate that activation of UGP1‐mediated heat production in brown adipose tissue during cold exposure reduces the necessity for shivering and thus prevents the development of severe dysfunction in shivering muscles.— Aydin, J., Shabalina, I. G., Place, N., Reiken, S., Zhang, S.‐J., Bellinger, A. M., Nedergaard, J., Cannon, B., Marks, A. R., Bruton, J. D., Westerblad, H. Nonshivering thermogenesis protects against defective calcium handling in muscle. FASEB J. 22, 3919–3924 (2008)

Ryanodine receptor fragmentation and sarcoplasmic reticulum Ca2+ leak after one session of high-intensity interval exercise

Significance High-intensity interval training (HIIT) has become popular because it is a time-efficient way to increase endurance. An intriguing and so-far-unanswered question is how a few minutes of HIIT can be that effective. We exposed recreationally active men to one session of three to six sets of 30-s high-intensity cycling exercise. Muscle biopsies taken 24 h later showed an extensive fragmentation of the sarcoplasmic reticulum (SR) Ca2+ channels, the ryanodine receptor 1 (RyR1). In isolated mouse muscle fibers, this fragmentation was accompanied by increased SR Ca2+ leak, which can trigger mitochondrial biogenesis. The HIIT-induced RyR1 fragmentation did not occur in muscles exposed to antioxidant, which offers an explanation for why antioxidants blunt effects of endurance training. High-intensity interval training (HIIT) is a time-efficient way of improving physical performance in healthy subjects and in patients with common chronic diseases, but less so in elite endurance athletes. The mechanisms underlying the effectiveness of HIIT are uncertain. Here, recreationally active human subjects performed highly demanding HIIT consisting of 30-s bouts of all-out cycling with 4-min rest in between bouts (≤3 min total exercise time). Skeletal muscle biopsies taken 24 h after the HIIT exercise showed an extensive fragmentation of the sarcoplasmic reticulum (SR) Ca2+ release channel, the ryanodine receptor type 1 (RyR1). The HIIT exercise also caused a prolonged force depression and triggered major changes in the expression of genes related to endurance exercise. Subsequent experiments on elite endurance athletes performing the same HIIT exercise showed no RyR1 fragmentation or prolonged changes in the expression of endurance-related genes. Finally, mechanistic experiments performed on isolated mouse muscles exposed to HIIT-mimicking stimulation showed reactive oxygen/nitrogen species (ROS)-dependent RyR1 fragmentation, calpain activation, increased SR Ca2+ leak at rest, and depressed force production due to impaired SR Ca2+ release upon stimulation. In conclusion, HIIT exercise induces a ROS-dependent RyR1 fragmentation in muscles of recreationally active subjects, and the resulting changes in muscle fiber Ca2+-handling trigger muscular adaptations. However, the same HIIT exercise does not cause RyR1 fragmentation in muscles of elite endurance athletes, which may explain why HIIT is less effective in this group.

Interpolated twitches in fatiguing single mouse muscle fibres: implications for the assessment of central fatigue

An electrically evoked twitch during a maximal voluntary contraction (twitch interpolation) is frequently used to assess central fatigue. In this study we used intact single muscle fibres to determine if intramuscular mechanisms could affect the force increase with the twitch interpolation technique. Intact single fibres from flexor digitorum brevis of NMRI mice were dissected and mounted in a chamber equipped with a force transducer. Free myoplasmic [Ca2+] ([Ca2+]i) was measured with the fluorescent Ca2+ indicator indo‐1. Seven fibres were fatigued with repeated 70 Hz tetani until 40% initial force with an interpolated pulse evoked every fifth tetanus. Results showed that the force generated by the interpolated twitch increased throughout fatigue, being 9 ± 1% of tetanic force at the start and 19 ± 1% at the end (P < 0.001). This was not due to a larger increase in [Ca2+]i induced by the interpolated twitch during fatigue but rather to the fact that the force–[Ca2+]i relationship is sigmoidal and fibres entered a steeper part of the relationship during fatigue. In another set of experiments, we observed that repeated tetani evoked at 150 Hz resulted in more rapid fatigue development than at 70 Hz and there was a decrease in force (‘sag’) during contractions, which was not observed at 70 Hz. In conclusion, the extent of central fatigue is difficult to assess and it may be overestimated when using the twitch interpolation technique.