Development of various types of muscle fibers in the quadriceps femoris and the soleus during human ontogenesis

By means of histological methods for revealing adenosine triphosphatase of myosin (pH 4.6) and succinate dehydrogenase activity, using postmortem material, development of various muscle fibers of the femoral m. quadriceps and m. soleus has been studied in human ontogenesis. The first stage of rearrangements lasts from the 5th-6th month of the uterine development up to 2 years of age and is characterized by formation (from non-differentiated) of oxidative, glycolytic and oxidative-glycolytic fibers. During the period from 2 up to 7-8 years of age the ratio in the types changes slightly, but transversal section size of the muscle fiber increases intensively. Then from 11 up to 17 years of age, together with maximal increment of the fibers transversal section, there is an essential change in the type relation. By the 17th years of age, in the femoral m. quadriceps the part of the fibers with glycolytic type of energy supply increases, while in the m. soleus the oxidative fibers become more numerous. By the 70th years of age in the femoral m. quadriceps relative amount of intermediate fibers increases.     

Mechanomyography of the human quadriceps muscle during incremental cycle ergometry

The mechanical activity of the human quadriceps muscle during maximal incremental cycle ergometry was investigated by mechanomyography (MMG). MMG and surface electromyography (EMG) recordings of vastus lateralis muscle activity were obtained from nine males. Cycle ergometry was performed at 60 rev/min and work load was incremented step wise by 20 W (3.2 Nm) every minute until volitional fatigue. The mean amplitudes of MMG (mMMG) and EMG (mEMG) during the contraction phase were calculated from the last six contractions in each load. The duration, load and work rate of exercise at exhaustion were 13.3 (1.6) min, 44.1 (5.5) Nm, 276.7 (34.7) W, respectively. A linear relationship between mMMG and load was evident in each subject (r = 0.868–0.995), while mEMG seemed to dissociate as the load became greater. In the grouped mean data, mMMG was linearly related to load whether aligned to the absolute (r = 0.995) or maximal (r = 0.995) load. Involvement of the noise component was further investigated by studying passive cycling by four subjects. Pedals were rotated passively for the first half of each stage (PAS) and the subject then pushed the pedals for the second half (ACT). In the lighter load region, the mMMG of ACT was as small as that of PAS. However, the change in the mMMG of PAS was very small compared with that of ACT. In conclusion, this study demonstrates a linear relationship between the mMMG of the quadriceps muscle and work load during maximal incremental cycle ergometry. The effect of movement noise was thought to be small and stable. Accepted: 22 April 1997     

Ultrastructural changes in the red muscle fibers of the quadriceps muscle of the rat femur during increased motor activity

Experiments on rats were made to study the effect of physical exercise running in a treadbahn on ultrastructure of skeletal muscle fibers. The biphasic mechanism of muscle contractile activity was shown. The processes of destruction occurring in red muscle fibers of the intensely working quadriceps femoris were manifested by enlargement of T system and sarcoplasmic reticulum elements, mitochondrial matrix swelling, cryst destruction, vacuolar degeneration of part of the mitochondria, and destruction of individual myofibrils. In addition to destructive changes, the muscle exhibited the recovery processes--physiological regeneration. Those processes included large accumulations of the mitochondria beneath the plasma membrane of muscle fibers, the presence of small mitochondria, division of the mitochondria, transformation of myosatellitocytes to myoblasts, the presence of centrioles in endotheliocytes, and so forth.     

Biphasic expression of slow myosin light chains and slow tropomyosin isoforms during the development of the human quadriceps muscle

Using a two-dimensional electrophoresis technique coupled with sensitive silver staining, we have investigated the chronology of appearance of the myosin light chain and tropomyosin isoforms during early stages of human quadriceps development. Our results show that slow myosin light chains and the slow tropomyosin isoform are not detected at 6 weeks of gestation. These isoforms transiently appear between 12.5 weeks and 15 weeks of gestation and then disappear. The slow myosin light chains are re-expressed at 31 weeks of gestation and the slow tropomyosin isoform later at 36 weeks of gestation, and normally remained expressed into the adulthood. Our study thus reveals a biphasic expression of the slow myosin light chains and the slow tropomyosin isoform in developing human quadriceps muscle.     

Rate of fatigue during repeated submaximal contractions of human quadriceps muscle.

Our purpose was to determine the effect of eight different combinations of contraction intensity, duration, and rest on the rate of fatigue in vastus lateralis muscle. A single combination consisted of contractions at 30 or 70% maximal voluntary contraction (MVC), held for 3 or 7 s with 3- or 7-s rest intervals. Contractions were repeated until the subject could not hold the force for the requisite duration. At regular intervals during each experiment, a brief MVC, a single twitch, and the response to eight stimulation pulses at 50 Hz were elicited. The rate of fatigue was the rate of decline of MVC calculated from regression analysis. Mean rate of fatigue (n = 8) ranged from 0.3 to 25% MVC/min and was closely related (r = 0.98) to the product of the relative force and the duty cycle. Force from 50 Hz stimulation fell linearly and in parallel with MVC. Twitch force was first potentiated and then fell twice as fast as 50 Hz stimulation and MVC (p less than 0.05). Differentiated twitch contraction and relaxation rates were higher at potentiation and lower at the limit of endurance, compared with control values (p less than 0.05). The maximal electromyogram decreased 25% and the submaximal EMG increased to maximal by the end of the protocol, indicating that the entire motor unit pool had been recruited. The close relation between rate of fatigue and the force x time product probably reflects the off-setting interaction of contraction amplitude, duration, and rest interval. This occurs despite the changes in twitch characteristics and the apparent recruitment of fast fatiguing motor units.     

Characteristics of the muscle mechanoreflex during quadriceps contractions in humans

We examined muscle sympathetic nerve activity (MSNA) in thenonexercising lower limb during repetitive static quadricepscontraction paradigm at 25% maximal voluntary contraction in eightmen. Subjects performed 20-s contractions with 5-s rest periods for upto 12 contractions. Although the workload was constant, we found that MSNA amplitude rose as a function of contraction number [0.6 ln (amplitude/min)/contraction]; this suggests chemicalsensitization of the muscle reflex response. We employedsignal-averaging techniques and then integrated the data to examine theonset latency of the MSNA response as a function of the 25-scontraction-rest period. We observed an onset latency of ~4-6 s.Moreover, although the onset latency did not appear to vary as afunction of contraction number, the rate of MSNA increase tookapproximately four contractions to reach a steady-state rate of rise;this suggests contraction-induced sensitization. The onset latencyreported here is similar to findings in recent animal studies, but itis at odds with latencies determined in prior human handgripcontraction studies. We believe our data suggest that1) mechanically sensitive afferentscontribute importantly to the MSNA response to the paradigm employedand 2) these afferents may besensitized by the chemical products of muscle contraction.     

Voluntary activation level and muscle fiber recruitment of human quadriceps during lengthening contractions.

Voluntary activation levels during lengthening, isometric, and shortening contractions (angular velocity 60 degrees/s) were investigated by using electrical stimulation of the femoral nerve (triplet, 300 Hz) superimposed on maximal efforts. Recruitment of fiber populations was investigated by using the phosphocreatine-to-creatine ratio (PCr/Cr) of single characterized muscle fibers obtained from needle biopsies at rest and immediately after a series of 10 lengthening, isometric, and shortening contractions (1 s on/1 s off). Maximal voluntary torque was significantly higher during lengthening (270 +/- 55 N.m) compared with shortening contractions (199 +/- 47 N.m, P < 0.05) but was not different from isometric contractions (252 +/- 47 N.m). Isometric torque was higher than torque during shortening (P < 0.05). Voluntary activation level during maximal attempted lengthening contractions (79 +/- 8%) was significantly lower compared with isometric (93 +/- 5%) and shortening contractions (92 +/- 3%, P < 0.05). Mean PCr/Cr values of all fibers from all subjects at rest were 2.5 +/- 0.6, 2.0 +/- 0.7, and 2.0 +/- 0.7, respectively, for type I, IIa, and IIax fibers. After 10 contractions, the mean PCr/Cr values for grouped fiber populations (regardless of fiber type) were all significantly different from rest (1.3 +/- 0.2, 0.7 +/- 0.3, and 0.8 +/- 0.6 for lengthening, isometric, and shortening contractions, respectively; P < 0.05). The cumulative distributions of individual fiber populations after either contraction mode were significantly different from rest (P < 0.05). Curves after lengthening contractions were less shifted compared with curves from isometric and shortening contractions (P < 0.05), with a smaller shift for the type IIax compared with type I fibers in the lengthening contractions. The results indicate a reduced voluntary drive during lengthening contractions. PCr/Cr values of single fibers indicated a hierarchical order of recruitment of all fiber populations during maximal attempted lengthening contractions.     

Distribution of myosin mRNA during development and regeneration of skeletal muscle fibers

Myosin mRNA distribution among subcellular compartments of anterior tibialis muscles in rabbit is monitored by in situ hybridization. A high density of mRNA was widely distributed throughout myotubes from 29-day fetal muscle and from regenerating adult muscle. All cytoplasmic spaces contained mRNA except where scattered myofibrils and centrally located nuclei were found. In fibers from 22-week-old rabbits, myosin mRNA was concentrated under the sarcolemma and excluded from the consolidated myofibrils and peripheral nuclei. The dispersal of mRNA through the cytoplasm in myotubes suggests that translation of myosin is widespread and that rapid myofibril assembly can occur throughout the fiber.     

Activation of human quadriceps femoris muscle during dynamic contractions: effects of load on fatigue.

Muscle fatigue is both multifactorial and task dependent. Electrical stimulation may assist individuals with paralysis to perform functional activities [functional electrical stimulation (FES), e.g., standing or walking], but muscle fatigue is a limiting factor. One method of optimizing force is to use stimulation patterns that exploit the catchlike property of skeletal muscle [catchlike-inducing trains (CITs)]. Although nonisometric (dynamic) contractions are important parts of both normal physiological activation of skeletal muscles and FES, no previous studies have attempted to identify the effect that the load being lifted by a muscle has on the fatigue produced. This study examined the effects of load on fatigue during dynamic contractions and the augmentation produced by CITs as a function of load. Knee extension in healthy subjects was electrically elicited against three different loads. The highest load produced the least excursion, work, and average power, but it produced the greatest fatigue. CIT augmentation was greatest at the highest load and increased with fatigue. Because CITs were effective during shortening contractions for a variety of loads, they may be of benefit during FES applications.     

Demonstration of ATPase activity in skeletal muscle fibers of the triceps muscle of arm in white rats during development

On the base of modified calcium and method from Zebe and Falk (1964) for demonstration of ATPase activity results at the long head of the triceps muscle of white rats aged 1, 3, 6 weeks and 3 month were shown. This method is proper to make visible alteration of the localisation of the reaction product during the postnatal development. Further investigations are necessary in order to correlate the steps of differentiation of muscle fibres with the strength of ATPase activity.     

Biomechanics of human quadriceps muscles during electrical stimulation

The quadriceps muscles of neurologically intact and spinal cord injured (SCI) human subjects were stimulated with constant current pulses. Up to three, separately adjustable stimulating electrodes over the motor points for vastus medialis (VM), vastus lateralis (VL) and rectus femoris (RF) muscles were used to maximize torque generation while minimizing discomfort. The torque generated by stimulation increased as the knee was slowly flexed to about 1 rad (50-60 degrees) and decreased beyond that point (a 'negative slope' on a torque-angle curve). Despite this region of negative slope the force generated by small oscillations remained positively correlated to the angle changes. When the knee was slowly extended again from a flexed position, the torque continued to decline and therefore showed a large degree of 'hysteresis'. Of the three heads studied, only stimulation of RF muscle generally produced this behavior. VL and VM had torques that increased monotonically with knee flexion over the range studied. The torques generated with electrical stimulation of normal subjects represented up to about 30% of maximum voluntary contraction. When subjects generated similar torques voluntarily, the negative slope region and substantial hysteresis were not observed. Thus, SCI subjects may be adversely affected by hysteresis during electrically-induced transitions from sitting to standing and vice versa, while normal subjects are not.     

Differentiation of muscle fibers of various types during postnatal development of the ventral serrate muscle in a rat]

In the denticulate ventral muscle of Wistar rats at the age of 1 day--2 months activity of NAD-N-dehydrogenase, succynic dehydrogenase and cytochrome oxidase has been determined in transversal cryostat sections. Quantitative estimation of the enzymes activity has been carried on by the plag-method. With age, general tendency to increasing activity of the enzymes mentioned is noted, but the dynamics of the increase is peculiar for every enzyme. Analysing the histograms on muscular fibre distribution according to their optical density, it is possible to estimate the dynamics. Simultaneously, the width of variational series, central statistical moments, indices of asymmetry and excess are also estimated. The whole course of the muscular fibre development, in accordance to the range and moments of distribution, can be devided into four main stages: stable, initial stage, slow increase of events, rapid increase of events and stabilization of the process. The stages mentioned pass gradually one into another making it possible to mark transitional stages (5--11, 15--19, 34--60 days). Using standard indices, it is possible to characterize more strictly the processes occurring in the course of muscular fibre differentiation. Lack of parallelism in the dynamics of asimilarity and excess can be treated as variety in differentiation of muscular fibres with middle and large optic density, and parallelism in dynamics--as their simultaneous differentiation. By comparing the curves it can be concluded that up to the 12--14th days, variety in differentiation of muscular fibres occurs, while after the 14th day their differentiation is more regular and simultaneous. The method of cytophotometry with subsequent mathematical processing of the results helps to determine the stages of muscular fibres differentiation.     

Mechanomyogram from the different heads of the quadriceps muscle during incremental knee extension

To investigate the time- and frequency-domain responses of mechanomyograms (MMGs) during the progressive fatigue induced by intermittent incremental contractions, a surface MMG was obtained from the three muscle heads of the quadriceps muscle in seven subjects while they performed isometric knee extensions lasting 7.6 min. Isometric intermittent incremental contractions started at 1% of the maximal voluntary contraction (MVC) for 3 s, with a 3-s relaxation period in between each contraction, and the contraction level was increased by 1% of MVC for every contraction (by 10% of MVC per min) up to exhaustion. Separate contractions with sufficient rest periods were also conducted to serve for the MMG characteristics without fatigue. The integrated MMG (iMMG) was linearly related to force in all of the muscles when fatigue was not involved. With regard to the incremental contractions, the relationship exhibited an ascending-descending shape, but the behavior was not the same for the individual muscle heads, especially for the rectus femoris muscle. A steep increase in the median frequency of MMG from around 60% of MVC corresponded to a decrease in iMMG. These results suggest that analysis of MMG in the time- and frequency-domain during an incremental protocol is a useful way of characterizing the motor unit recruitment strategy and fatigue properties of individual muscles. Accepted: 19 March 1998     

Microphotometric determination of glycogen in single fibres of human quadriceps muscle

Synopsis A technique for the quantitation of glycogen in single fibres of human skeletal muscle is described. By using microphotometry the loss of glycogen from cryostat sections during a PAS-staining procedure was shown to be negligible. Further, it was found that nearly all the PAS-positive material (98.5%) inside a muscle fibre is glycogen. A significantly higher mean glycogen concentration (P<0.001) was found in type II fibres than in type I fibres in the resting quadriceps muscle of sedentary young males. The coefficient of variation for the glycogen concentration within each fibre type was found to be 17% and 15% for type I and type II respectively. The specificity of the PAS-staining technique for glycogen was confirmed by a statistically significant correlation (r=0.78,P<0.001) between the glycogen concentration measured biochemically and that calculated from microphotometry and area and thickness measurements. With the technique described, it seems possible to measure the glycogen concentration of single muscle fibres in serial sections and to calculate this in standard biochemical terms.     

Effect of arterial oxygenation on quadriceps fatigability during isolated muscle exercise

The effect of various levels of oxygenation on quadriceps muscle fatigability during isolated muscle exercise was assessed in six male subjects. Twitch force (Q(tw)) was assessed using supramaximal magnetic femoral nerve stimulation. In experiment 1, maximal voluntary contraction (MVC) and Q(tw) of resting quadriceps muscle were measured in normoxia [inspired O(2) fraction (Fi(O(2))) = 0.21, percent arterial O(2) saturation (Sp(O(2))) = 98.4%, estimated arterial O(2) content (Ca(O(2))) = 20.8 ml/dl], acute hypoxia (Fi(O(2)) = 0.11, Sp(O(2)) = 74.6%, Ca(O(2)) = 15.7 ml/dl), and acute hyperoxia (Fi(O(2)) = 1.0, Sp(O(2)) = 100%, Ca(O(2)) = 22.6 ml/dl). No significant differences were found for MVC and Q(tw) among the three Fi(O(2)) levels. In experiment 2, the subjects performed three sets of nine, intermittent, isometric, unilateral, submaximal quadriceps contractions (62% MVC followed by 1 MVC in each set) while breathing each Fi(O(2)). Q(tw) was assessed before and after exercise, and myoelectrical activity of the vastus lateralis was obtained during exercise. The percent reduction of twitch force (potentiated Q(tw)) in hypoxia (-27.0%) was significantly (P < 0.05) greater than in normoxia (-21.4%) and hyperoxia (-19.9%), as were the changes in intratwitch measures of contractile properties. The increase in integrated electromyogram over the course of the nine contractions in hypoxia (15.4%) was higher (P < 0.05) than in normoxia (7.2%) or hyperoxia (6.7%). These results demonstrate that quadriceps muscle fatigability during isolated muscle exercise is exacerbated in acute hypoxia, and these effects are independent of the relative exercise intensity.     

Electrophysiological characteristics of muscle fibers during denervation

Experimental material obtained on mouse extensor digitorum longus was presented concerning the dynamics of changes in the membrane potential (MP) of muscle fibres after chronic denervation (1-12 days), the effect of ouabain and increase of extracellular potassium on MP of normal and denervated muscles, changes of input resistance and volumes of muscle fibres after denervation.