Longitudinal study of the contractile and electrical properties of single human thenar motor units

Serial motor unit number estimates have shed important light on the extent and rates of motoneuron losses in aging and amyotrophic lateral sclerosis. However, the estimates alone provide few clues to the health and functional status of surviving motor units. A reliable means for assessing the functional status of the surviving motor units would therefore by a welcome addition to our present tools for studying motor units. Examining the physiological properties of samples of motor units drawn at intervals during the course of a motoneuronal disease suffers from the important limitation that the samples may not be representative of one another. The latter problem could be circumvented by serially studying the same motor units. This study describes a noninvasive technique capable of longitudinally tracking the contractile and electrical properties of specific single thenar motor units in healthy subjects, in some instances over several years. The technique proved to be reasonably reliable and provided information on a wide range of contractile and electrical properties of motor units. Such an approach could serve as a potentially powerful and sensitive means of studying the life histories of single motor units in aging, diseases of the motoneuron, and in the latter instances, the responses of the motoneurons to treatment.     

Responses of human single motor units to transcranial magnetic stimulation

The authors analyse the results of 51 unicompartmental knee prostheses with 1 to 12 years follow-up (mean follow-up: 5 years). The results were evaluated using the scoring system of the "Knee Group" of the SO.B.C.O.T. (Société Belge de Chirurgie Orthopédique et de Traumatologie). This analysis demonstrates that the quality of the results depends on implant positioning. The authors suggest positioning the tibial implant parallel with the healthy plateau and slightly distal, i.e. to position the tibial implant perpendicular to the epiphyseal axis and not to the mechanical axis, as is systematically done with the usual tibial cutting guides. When this ideal positioning was respected, 77.5% of the patients had a score above 90 points (out of a possible maximum of 100 points) and 12.5% had a score between 75 and 89 points. When this condition was not respected, none of the knees obtained more than 75 points. The difference was statistically significant (p = 0.0001).     

Directional tuning of single motor units

The directional activity of whole muscles has been shown to be broadly and often multimodally tuned, raising the question of how this tuning is subserved at the level of single motor units (SMUs). Previously defined rules of SMU activation would predict that units of the same muscle (or at least of the same neuromuscular compartment) are activated homogeneously with activity peaks in the same "best" direction(s). In the present study, the best directions of SMUs in human biceps (both heads) and deltoid (anterior, medial, and posterior portions) were determined by measuring the firing rate and threshold force of units for recruitment during isometric force ramps in many different directions. For all muscles studied, neighboring motor units could have significantly different best directions, suggesting that each muscle receives multiple directional commands. Furthermore, 17% of the units sampled clearly had a second-best direction, consistent with a convergence of different directional commands onto the same motoneuron. The best directions of the units changed gradually with location in the muscle. Best directions did not cluster into separate groups, thus, not supporting the existence of clearly distinguished neuromuscular compartments. Instead, the results reveal a more gradually distributed activation of the biceps and deltoid motoneuron pools. A model is proposed in which the central control mechanism optimizes the fulfillment of the continuously changing directional force requirements of a movement by gradually recruiting and derecruiting those units ideally suited for the production of the required force vector at any given time.     

Mechanical properties and behaviour of motor units in the tibialis anterior during voluntary contractions

The present work was carried out to analyse the properties and behaviour of Tibialis anterior motor units (MUs) during voluntary contractions in humans. A total of 528 single MU mechanical properties was recorded in 10 subjects by means of the spike-triggered averaging (STA) technique. MU recruitment thresholds and discharge frequencies were recorded during linearly increasing maximal voluntary contraction (MVC). The results indicate a mean (+/- SD) MU torque of 25.5 +/- 21.5 mN.m. and a mean time-to-peak of 45.6 +/- 13.6 ms. A comparison of the average MU twitch torque with that of the muscle allowed an estimate of about 300 MUs in the Tibialis anterior. A positive linear relationship was recorded between the MU twitch torque and the recruitment threshold. The mean minimal and maximal discharge frequencies of MUs were 8.4 +/- 3.0 Hz and 33.2 +/- 14.7 Hz, respectively. The results of the present work indicate that MU behaviour during voluntary contractions is different in the tibialis anterior and in the adductor pollicis.     

Organization of single motor units in feline sartorius

1. We depleted single motor units in feline sartorius muscles of glycogen by stimulating their motoneurons intracellularly. We mapped the intramuscular distribution of depleted fibers by inspecting histological cross-sections throughout the length of sartorius. 2. We selected ten depleted motor units for detailed study and quantitative analysis. Nine motor units were located in the anterior head of sartorius. One was located in a muscle whose distal half appeared to have been damaged some time before the acute experiment. A single motor unit was located in the medial head of sartorius. 3. Five motor units were composed of fast-twitch glycolytic (FG) muscle fibers, two of fast-twitch oxidative glycolytic (FOG) muscle fibers, and three of slow-twitch oxidative (SO) muscle fibers. Estimates of the numbers of depleted fibers in motor units of anterior sartorius indicated that FG motor units were larger (mean 566 fibers) than FOG and SO motor units (SO mean 190, FOG mean 156 fibers). The SO motor unit in the damaged muscle had 550 fibers. One motor unit depleted in the medial head of sartorius had 270 fibers with FG profiles. 4. Muscle fibers belonging to each anterior motor unit were never distributed throughout the whole cross-section of anterior sartorius at any proximodistal level. Furthermore, fibers were distributed nonuniformly along the proximodistal axis of the muscle. In most muscles at least a few depleted fibers were found at all proximodistal levels. However, in one normal muscle and the damaged muscle, depleted fibers were confined to the proximal end. 5. The fibers in the medial motor unit were confined to a strip that did not extend across the whole cross-section of the muscle head. Fibers within this strip were scattered quite evenly from origin to insertion. This medial FG motor unit occupied a smaller territory and contained fewer fibers than anterior motor units of the same histochemical type. 6. These results show that sartorius motor units are not distributed uniformly in the mediolateral plane; those in anterior sartorius were distributed asymmetrically in the proximodistal axis as well. This finding has important functional implications for the way in which we model force development and transmission in sartorius and other long muscles.     

Discharge pattern of single motor units in the tonic vibration reflex of human triceps surae

Using a single fibre EMG electrode the firing pattern of 46 motor units in the triceps surae has been studied during vibration of the Achilles tendon at frequencies of 25--200 Hz. Potentials activated in the tonic vibration reflex (TVR) were phase-locked to the vibration cycle but tended to become somewhat less so with continued vibration. The firing pattern of voluntarily activated motor units became locked to the waveform by the application of the vibrator. The discharges of 21 motor units were studied during low threshold (sub-M wave) tetanic stimulation of the tibial nerve at 25--100 Hz. No evidence was found of synchronization of potentials activated in the resulting tonic contraction. During weak voluntary contractions, stimulation also failed to regularize voluntarily activated motor units. The findings can be reconciled by postulating that, in normal man, vibration activates monosynaptic and polysynaptic pathways, the latter circuit being adequate to generate reflex contraction, while the former merely affects the temporal patterning of the motor outflow.     

Sprint-training effects on some contractile properties of single skinned human muscle fibres

The effects of sprint training on the contractile properties of human muscle fibres obtained by needle biopsy were investigated. Individual fibres were mechanically skinned and activated by Ca(2+)- and Sr(2+)-buffered solutions at pH 7.1, and allocated to distinct populations on the basis of their contractile characteristics. The majority of fibres sampled pre-training could be separated into the three major fibre groups: Populations I (24/70, 34%), II (25/70, 36%) and III (18/70, 26%), which exhibited characteristics similar to those of histochemically classified type I, IIA and IIB fibres, respectively. The remainder (3/70, 4%) represented another fibre group, with intermediate characteristics. The muscle fibres were also activated by Ca2+ at a reduced pH of 6.6, to mimic the intracellular acidification that occurs during intense exercise. Lowering pH increased the threshold for contraction by Ca2+, reduced Ca2+ sensitivity, and increased the steepness of the force-pCa relationship, in all fibres sampled from the three major fibre groups. Maximum force was not significantly reduced in any fibre population. In the post-training sample, the three major fibre types were present in different proportions: Populations I (10/52, 19%), II (20/52, 38.5%) and III (11/52, 21%). Three other fibre groups sampled in low numbers exhibited contractile characteristics intermediate between Population I and Population II. Following sprint training all of the three main fibre populations exhibited higher thresholds for contraction by, and lower sensitivities to, Sr2+ but not Ca2+, compared with the fibres sampled pre-training. Maximum force was significantly lower in Population II fibres after sprint training. At pH 6.6, post-trained Population III fibres exhibited even lower Ca2+ sensitivity, with concomitant increases in the threshold for contraction and force-pCa curve steepness.     

A longitudinal study of the pathophysiological changes in single human thenar motor units in amyotrophic lateral sclerosis

The sequence of pathophysiological changes in amyotrophic lateral sclerosis (ALS) at the single motor unit (MU) level is not well understood. Using a recently described technique, a comprehensive range of physiological properties in two thenar MUs in ALS were intensively studied. In the first MU, despite a marked decline in the ability of the subject to voluntarily recruit the MU, the physiological properties of this MU remained remarkably stable over a 2-year period. In contrast, the physiological properties of the other MU declined rapidly over 5 months despite the fact that this MU could be recruited with ease throughout the study period. These differences between the progressively dysfunctional changes in these two MUs illustrates the value of such longitudinal studies of specific MUs in improving our understanding of the evolution of changes in single motoneurons in ALS. The broader application of longitudinally tracking the pathophysiological changes of the surviving MUs may prove to be a sensitive measure of disease progression and in evaluating the effectiveness of treatments.     

The contractile properties of the elderly human diaphragm

It has previously been reported that aging is associated with a substantial decrease in diaphragm strength. To test this hypothesis we studied 15 (10 male, 5 female) subjects with a mean age of 29 (range 21 to 40) and 15 elderly (10 male, 5 female) subjects, mean age 73 (range 67 to 81). We measured transdiaphragmatic pressure (Pdi) during a maximal sniff (Sniff Pdi) and during bilateral cervical magnetic stimulation (CMS) of the phrenic nerve roots (Tw Pdi). Additionally in 17 subjects (9 elderly and 8 young) the Pdi elicited by paired CMS (pTw Pdi) was obtained at interstimulus intervals ranging from 10 to 999 ms (1 to 100 Hz). There was considerable overlap between groups. Mean Sniff Pdi in the elderly was 119 cm H2O compared with 136 cm H2O for the young subjects; this represented a median reduction of 18 cm H2O or 13% (p = 0.05, 95% Cl of difference 0 to 33 cm H2O). Mean Twitch Pdi in the elderly was 26.8 cm H2O compared with 35.2 cm H2O, a median reduction of 8 cm H2O or 23% (p = 0.004, 95% Cl 3 to 13 cm H2O). At 10 Hz the elderly tended to generate a higher fraction of the Pdi obtained at 100 Hz than the young, but this trend did not achieve statistical significance (p = 0.11). We conclude that aging is associated with a reduction in diaphragm strength. However the magnitude of the reduction is small and may be offset by a leftward shift of the force-frequency relationship.     

Recruitment stability in masseter motor units during isometric voluntary contractions

Recruitment of single motor units (SMUs) of the masseter muscle was studied using macro representation (MacroRep) as the indicator of motor unit size. When subjects followed a slow isometric force ramp, units were usually recruited in order of MacroRep size. However, pooling the data from repeated ramps in the same subject resulted in a weak relationship between MacroRep size and force recruitment threshold, probably due to marked variations in the relative contributions of the jaw muscles, and varying levels of cocontraction, in the development of total bite force in each ramp. The force recruitment thresholds of individual SMUs showed marked variability, but recruitment threshold stability was improved when expressed as a percentage of maximum surface electromyographic (SEMG) activity in the ipsilateral masseter. Therefore the SEMG recruitment threshold was concluded to be a more stable and accurate indicator of the SMU's position in the recruitment hierarchy in a given muscle. It was concluded that SMUs in masseter are recruited according to the size principle, and that when investigating recruitment in jaw muscles, SEMG recruitment threshold should be used in preference to force recruitment threshold.     

Response properties of single units in an association area of the kitten neocortex

Two percept-genetic methods, the Meta-Contrast Technique (MCT) and the Serial Afterimage Test (AI), were used to study adaptive aspects of perception in a group of 37 subjects suffering from manifest anxiety. The perceptual processes laid bare by these methods reflected not only anxiety but also defensive formations. MCT and AI correlated with each other in these respects and, when combined, correlated with rated symptoms. In accordance with the theory behind percept-genetic methods, anxiety signs appearing late in the test serials were more likely to correspond to manifest symptoms than signs which appeared early. The AI test scores seemed to reflect a difference between anxiety experienced as an inherent quality of life and anxiety as a foreign force struggling for dominance.     

Gender differences in isometric contractile properties and fatigability in elderly human muscle

This study had two purposes: to compare the isometric contractile characteristics and fatigability in the elbow flexors (EF) and ankle dorsiflexors (DF) in older males and females (60-80 years), and to determine the effects of almost 2 years of twice weekly dynamic weight-lifting training on these properties One hundred nine male and female subjects completed the 22-month intervention, 54 in the exercise group and 55 in the control group. Isometric contractile properties and fatigue characteristics were assessed at baseline, 10 months, and 22 months. At baseline, males were significantly stronger (p < .05) than females in evoked twitch torque (Pt) and maximal voluntary strength (MVC) in both EF and DF. Females exhibited significantly less fatigue (expressed either absolutely or relatively) than males in both Pt and MVC during a 3-min protocol of intermittent MVCs (5 s contraction; 2 s rest). There were no significant effects of the 22 months of dynamic training on either the isometric contractile properties or the fatigue characteristics, suggesting a very strong and persistent specificity of training effect.     

On the preservation of contractile proteins during storage of human platelets

A study has been undertaken to determine the rate at which stored platelets lose their ability to respond to stimuli and to establish whether this decrease in function could be ascribed to the storage-induced proteolysis of prominent platelet proteins observed by others. Platelet concentrates were stored at 4 degrees C and 25 degrees C for up to 14 days, and their ability to secrete and aggregate in response to appropriate stimuli was determined at 6, 96, and 192 hr after venipuncture. At each time point the protein complement of the platelets was also monitored by SDS-polyacrylamide gel electrophoresis to assess the extent of intracellular protein degradation. Platelets from concentrates stored at either temperature exhibited a decreased ability to respond to stimuli as storage time increased. After 8 days of storage at 4 degrees C and up to 9 days at 25 degrees C, no proteolysis of major platelet proteins was observed; however, complete loss of platelet function was observed. This strongly indicates that a decrease in platelet function should not be causally linked to degraded contractile-structural proteins and that extending the functional life of platelets during storage is still an attainable goal since proteolysis is not the inevitable result of short-term storage.     

The reorganization of motor units in motor neuron disease

We studied 78 patients with motor neuron disease (MND) using concentric needle electromyography. Analysis on weak and maximal effort was performed using our own, fully automated, computer method, EMG-LAB. In addition to the conventional parameters of single motor unit action potentials (MUAPs) and interference pattern, new criteria were applied: the range of the acting motor units and the functional recruitment order. A total of 375 muscles of MND patients and 120 control muscles were investigated. The electromyographic data were analyzed separately in five groups of muscles, classified A, B, C, D, and E according to their clinical condition. Those results allowed us to discern six neurophysiological stages (N(0,1,2,3,4,5)) from the early to the most advanced phase. It has been confirmed that reinnervation in MND is adequate to compensate for the loss of over 50% of motor neurons but it is only a transitory phase in the morbid course. At stages N(O-5), the electrophysiological data reflect structural and functional integrity of the functioning motor units. Evaluation of not only single MUAPs but also of the full range of acting motor units and their recruitment order allowed a deeper look into the underlying pathophysiological mechanisms.     

Chronic administration of taurine to aged rats improves the electrical and contractile properties of skeletal muscle fibers

Fimbriae or pili are essential adherence factors usually found in pathogenic bacteria to aid colonization of host cells. Three major structural pilin genes, fimA, sfaA, and papA, from Escherichia coli natural isolates were examined and nucleotide sequence data revealed elevated levels of both synonymous and nonsynonymous site variation at these loci. Examination of synonymous site variation shows a fivefold increase in fimA sites, relative to the housekeeping gene mdh; and similarly the sfaA and papA genes have increased synonymous sites variation relative to fimA. Nonsynonymous site variation is also elevated at all three loci but, in particular, at the papA locus (kN = 0.44). The kN/kS ratio for the three genes are among the highest yet reported for E. coli genes. Regional variation in nucleotide polymorphism within each of the genes reveal hypervariable segments where nonsynonymous substitutions exceed synonymous substitutions. We propose that at the fimA, papA, and sfaA genes, diversifying selection has brought about the increase levels of polymorphism.     

Frequency domain electrical conductivity measurements of the passive electrical properties of human lymphocytes

Use of liposomes as carriers of gentamicin for intrabronchial pulmonary delivery was investigated in rabbits. Gentamicin, in isotonic glutamic acid buffer, pH 4.5, was encapsulated in multilamellar vesicles (MLVs) and administered intrabronchially. Higher drug concentrations were found at the pulmonary site of liposome instillation for 1 day as compared with free unencapsulated antibiotic. When time-course distributions of gentamicin given in the liposomal or free form were measured in bronchoalveolar lavages (BAL), similar accumulations were observed up to 4 h, but the drug remained longer (24 h) after administration of the liposomal formulation. Higher amounts of antibiotic were detected in BAL supernatant 1 h after instillation of plain gentamicin; this difference stopped being significant after 4 h. A microbiological assay outlined the bacteriostatic activity of gentamicin released from MLVs and recovered in BAL supernatant. Liposomal gentamicin accumulated in the BAL cell pellet 1 h after intrabronchial instillation; it decreased progressively but minute amounts were still detected after 1 day. On the contrary, no gentamicin was found in the pellet at any time after free drug administration. Comparison of aminoglycoside concentrations in plasma and kidneys indicated lower and constant levels when the liposomal form was instilled. Liposome encapsulation altered the disposition of gentamicin in a way suggesting improved pulmonary concentration and lower systemic toxicity.     

N-acetylcysteine depresses contractile function and inhibits fatigue of diaphragm in vitro

We have previously shown that antioxidant enzymes (superoxide dismutase and catalase) depress contractility of unfatigued diaphragm fiber bundles and inhibit development of acute fatigue. In the present study, we tested for similar effects of N-acetyl-cysteine (NAC), a nonspecific antioxidant approved for clinical use. Diaphragms were excised from deeply anesthetized rats. Fiber bundles were removed, mounted isometrically at 37 degrees C, and stimulated directly using supramaximal current intensity. Studies of unfatigued muscle showed that 10 mM NAC reduced peak twitch stress (P < 0.0001), shortened time to peak twitch stress (P < 0.002), and shifted the stress-frequency curve down and to the right (P < 0.05). Fiber bundles incubated in 0.1-10 mM NAC exhibited a dose-dependent decrease in relative stresses developed during 30-Hz contraction (P < 0.0001) with no change in maximal tetanic (200 Hz) stress. NAC (10 mM) also inhibited acute fatigue. Throughout 10 min of intermittent contraction at 30-40 Hz, treated bundles developed higher stresses than time-matched control bundles (P < 0.0001). NAC concentrations > or = 30 mM were toxic, causing a prompt irreversible decrease in maximal tetanic stress (P < 0.0001). Because NAC effects mimic the effects of other antioxidant agents with different mechanisms of action, we conclude that exogenous antioxidants exert stereotypical effects on contractile function that differ between unfatigued and fatiguing muscle. Unlike antioxidant enzymes, however, NAC has been approved for clinical use and may be used in future studies of human muscle fatigue.