Combined in situ analysis of metabolic and myoelectrical changes associated with electrically induced fatigue.

Electrical muscle stimulation (Mstim) at a low or high frequency is associated with failure of force production, but the exact mechanisms leading to fatigue in this model are still poorly understood. Using 31P magnetic resonance spectroscopy (31PMRS), we investigated the metabolic changes in rabbit tibialis anterior muscle associated with the force decline during Mstim at low (10 Hz) and high (100 Hz) frequency. We also simultaneously recorded the compound muscle mass action potential (M-wave) evoked by direct muscle stimulation, and we analyzed its post-Mstim variations. The 100-Hz Mstim elicited marked M-wave alterations and induced mild metabolic changes at the onset of stimulation followed by a paradoxical recovery of phosphocreatine (PCr) and pH during the stimulation period. On the contrary, the 10-Hz Mstim produced significant PCr consumption and intracellular acidosis with no paradoxical recovery phenomenon and no significant changes in M-wave characteristics. In addition, the force depression was linearly linked to the stimulation-induced acidosis and PCr breakdown. These results led us to conclude that force failure during 100-Hz Mstim only results from an impaired propagation of muscle action potentials with no metabolic involvement. On the contrary, fatigue induced by 10-Hz Mstim is closely associated with metabolic changes with no alteration of the membrane excitability, thereby underlining the central role of muscle energetics in force depression when muscle is stimulated at low frequency. Finally, our results further indicate a reduction of energy cost of contraction when stimulation frequency is increased from 10 to 100 Hz.

Effects of endogenous nitric oxide in activation of group IV muscle afferents.

Based on previous observations that acute hypoxemia, which enhances nitric oxide (NO) production, depresses the activation of group IV afferents after repetitive low-frequency muscle stimulation (MS), we hypothesized that endogenous NO modulates the response of these nerve endings to their specific stimuli. The present study in rabbits examined the effects of a blocker of NO synthase (NG-nitro-L-arginine methyl ester L, L-NAME) and an exogenous NO donor (3-morpholinosydnonimine, SIN-1) on the group IV afferents of tibialis anterior. The efficacy of the two test agents was judged by their effects on systemic blood pressure. L-NAME markedly elevated (+46%) the resting discharge rate of group IV afferents but abolished their activation after repetitive MS. After SIN-1 injection, there was a transient decrease in blood pressure, which correlated well with a lowered resting discharge rate of group IV afferents. SIN-1 infusion caused a stable reduction of blood pressure; the resting afferent nerve discharge rate began first to decrease but then recovered control mean values. SIN-1 infusion abolished the activation of group IV afferents after MS. This study indicates that endogenous NO production in a resting or contracting muscle attenuates the baseline activity of group IV muscle afferents and their activation after repetitive muscle contractions.

Changes in afferent activities from tibialis anterior muscle after nerve repair by self-anastomosis.

In order to study sensory nerve plasticity after nerve injury and repair, recordings were made from afferent axons innervating the tibialis anterior muscle in rats under several different experimental conditions. In two groups of rats, reinnervation of the denervated tibialis anterior was examined 2.5 months (group A) and 7 months (group B) after section, along with self-anastomosis of the common peroneal nerve. The other rats (group C) were examined 2.5 months after the nerve was cut and ligatured to its stumps to avoid axonal regeneration. No evoked potentials and no activation in response to any test agent were found in group C rats. We found a significant increase in the proportion of group I-II fibers and a significant decrease in group IV fibers in the group B rats when compared with group A (P < 0.05 and P < 0.01) and control animals (P < 0.01 and P < 0.01). A higher conduction velocity was measured in group IV fibers in group B rats when compared with group A (P < 0.01) and the controls (P < 0.01). The proportion of afferent units showing an optimal discharge in response to tendon vibration at 70 Hz (range 0-100 Hz) was higher in groups A and B (72.2 and 80%, respectively) than in the controls (36.8%). The response of muscle afferents to KCl (1-20 mM) and lactic acid (0.5-3 mM) concentrations was markedly depressed in group A rats (P < 0.05), whereas it was restored and even accentuated in group B animals when compared with the controls (P < 0.05). Electrically induced fatigue (3 min, 10 Hz) significantly activated (P < 0.05) muscle afferents only in controls. The present study indicates that after self-anastomosis of a cut hindlimb muscle nerve, sensory innervation was markedly modified in the direction of enhanced mechanosensitivity to high-frequency tendon vibration and depressed metabosensitivity.

Influence of oxygen supply on activation of group IV muscle afferents after low-frequency muscle stimulation.

Anaerobic muscle metabolism and local release of inflammatory mediators play key roles in the mechanism of postfatigue-induced activation of group IV muscle afferents. The present study focused on activation of these muscle afferents after a 3-min period of low-frequency muscle stimulation (LFMS) in different conditions of muscle oxygenation, such as occur in patients with respiratory insufficiency and subjects living at high altitude. In anesthetized rabbits, spontaneous activity of group IV afferents (conduction velocity = 1.52 +/- 0.13 m.s(-1)) from the tibialis anterior muscle was recorded at rest (baseline) and then after LFMS under normoxic (PaO(2) = 113 mmHg), hyperoxic (PaO(2) = 186 mmHg), or hypoxic (PaO(2) = 35 mmHg) conditions. The maximal force decay at the end of LFMS did not differ significantly with respect to conditions of muscle oxygenation. Compared with normoxia, hypoxia significantly increased the baseline activity of group IV muscle afferents, whereas no effect was noted when hypoxia followed a period of hyperoxia. LFMS-induced activation of group IV afferents occurred in all circumstances, except when hypoxia was first tested. The activation of group IV muscle afferents after LFMS was markedly reduced when hypoxia followed normoxia (+14% versus +27%) or hyperoxia (+55% versus +144%), whereas it was accentuated when hyperoxia followed hypoxia (+25% versus +8%). We concluded that the sensorimotor control of skeletal muscles may be altered during acute hypoxia but facilitated after reoxygenation.

Mechanisms of fatigue-induced activation of group IV muscle afferents: the roles played by lactic acid and inflammatory mediators.

We already showed that group IV muscle afferents are activated during electrically-induced fatigue. The purpose of this study is to identify the mechanisms of stimulation of these muscle afferents by electrically-induced fatigue at a high (100 Hz; high frequency fatigue, HFF) or a low rate (10 Hz; low frequency fatigue, LFF) of stimulation. In 23 paralyzed and anaesthetized rabbits, group IV afferent activity from the tibialis anterior muscle was recorded before and after 3-min HFF or 5-min LFF runs eliciting the same force failure. Plasma lactic acid concentration (LA) was also measured in leg venous blood. We tested the effects of dichloroacetate (DCA), which reduces lactic acid production, and of acetylsalicylic acid (ASA), a blocker of cyclooxygenase, on fatigue-induced activation of group IV muscle afferents after HFF and LFF trials. Pretreatment by ASA or DCA did not modify HFF-induced activation of muscle afferents. On the other hand, LFF-induced response was markedly depressed by each pharmacological agent (-44% after ASA and -75% after DCA). We verified that DCA markedly lowered LA production after LFF. The present results show that the activation of group IV muscle afferents by LFF or HFF results from different mechanisms and also demonstrate the major role played by lactic acid production and, to a lesser extent, the release of inflammatory mediators in LFF-induced activation of group IV muscle afferents.

Modifications of afferent activities from Tibialis anterior muscle in rat by tendon vibrations, increase of interstitial potassium or lactate concentration and electrically-induced fatigue.

Although previous experiments with a partially similar objective have been described in dogs, cats and rabbits, the purpose of this study was to identify and characterize mechanosensitive and chemosensitive muscle afferents in the anaesthetized rat since it is a widely used laboratory animal. The peroneal nerve innervating the tibialis anterior muscle was studied. Measurement of conduction velocities from compound action nerve potentials evoked by peripheral nerve stimulation allowed identification of group I-II (10.79+/-1.02 m/s), group III (2.96+/-0.58 m/s) and group IV (0.46+/-0.07 m/s) afferent fibers. Computation of the different compound potential areas showed that afferents I and II arising from spindles and tendon organs represented 9.65+/-2.2%, whereas afferents III and IV arising from free nerve endings in muscle represented 90.35+/-2.2% (III, 46.66+/-2.71% and IV, 43.69+/-2.52%). Action potentials were recorded from teased nerve filaments. Mechanical tendon vibrations (10 to 90 Hz) were used to activate mechanoreceptors. Peak increase in afferent discharge (fimpulses) was measured at 50 Hz (n = 12/19 units) or 70 Hz (n = 7/19 units). Intra-arterial bolus injections of different concentrations of potassium chloride (KCl: 1 to 20 mM) or lactic acid (LA: 0.5 to 3 mM) elicited marked activation of III and IV afferents (n = 124). Enhancement of fimpulses was not proportional to the increase in [KCl] or [LA]. Activation of afferents plateaued when [KCl] was equal or greater than 5 mM while fimpulses peaked, then decreased, when [LA] was 1 mM. Muscle fatigue induced by direct electrical muscle stimulation (EIF) markedly activated group III-IV (n = 17/18) afferents (176.9+/-29.7% of control) which persisted for the 3 minutes of recovery from fatigue. Maximal fimpulses increases in response to LA (+67%) and KCl (+46.9%) injections and to EIF (+76.9%) were similar. This procedure for characterizing the functional properties of sensory nerve endings in a skeletal muscle may be used to assess further changes in sensory muscle paths in experimental rodent pathophysiological systems.

Fatigue-induced changes in group IV muscle afferent activity: differences between high- and low-frequency electrically induced fatigues.

Recordings of group IV afferent activity of tibialis anterior muscle were performed in paralysed rabbits during runs of electrically induced fatigue produced by direct muscle stimulation at a high (100 Hz, high-frequency fatigue HFF) or a low rate (10 Hz, low-frequency fatigue LFF). In addition to analysis of afferent nerve action potentials, muscle force and compound muscle action potentials (M waves) elicited by direct muscle stimulation with single shocks were recorded. Changes in M wave configuration were used as an index of the altered propagation of membrane potentials and the associated efflux of potassium from muscle fibers. The data show that increased group IV afferent activity occurred during LFF as well as HFF trials and developed parallel with force failure. Enhanced afferent activity was significantly higher during LFF (maximal delta f(impulses) = 249 +/- 35%) than HFF (147 +/- 45%). No correlation was obtained between the responses of group IV afferents to LFF or to pressure exerted on tibialis anterior muscle. On the other hand, decreased M wave amplitude was minimal with LFF while it was pronounced with HFF. Close correlations were found between fatigue-induced activation of group IV afferents and decreases in force or M wave amplitude, but their strength was significantly higher with LFF compared to HFF. Thus, electrically induced fatigue activates group IV muscle afferents with a prominent effect of low-frequency stimulation. The mechanism of muscle afferent stimulation does not seem to be due to the sole increase in extracellular potassium concentration, but also by the efflux of muscle metabolites, present during fatiguing contractions at low rate of stimulation.