Intrafibre distribution of succinate dehydrogenase in cat tibialis anterior motor units.

Using isolated ventral root filament stimulation and glycogen depletion techniques, 14 motor units from the cat tibialis anterior were studied. Based on their mechanical properties, the units were classified as either slow-fatigue resistant, fast-fatigue resistant, fast-fatigue intermediate, or fast-fatigable. Quantitative histochemical and computer assisted image analysis techniques were used to determine the activity of succinate dehydrogenase in a population of fibres in each unit. In addition, the intrafibre distribution of succinate dehydrogenase activity was measured in those same fibres by calculating the enzymatic activity of circumferential layers every 0.5 microns starting from the fibre edge to its centre. It was established that enzymatic activity and radial distance were linearly related in the fibres. A range in succinate dehydrogenase activity (mean coefficient of variation, 29%) was observed among the fibres of a unit. In contrast, the intrafibre distribution of that activity was rather consistent (mean variation, 4%) across the fibres of a unit. Further, the intrafibre distribution was similar among the fibres of units classified as the same type. However, the intrafibre distribution was disparate among the different unit types. These data suggest that the intrafibre distribution of mitochondrial enzymes may contribute to the mechanical properties of a motor unit. In this regard, a hypothesis is proposed that describes how the absolute activity of a mitochondrial enzyme, and the intrafibre distribution of that activity, may interactively contribute to the fatigue resistance of a unit.     

Interspecies differences of motor units properties in the medial gastrocnemius muscle of cat and rat

The purpose of the study was to analyze the interspecies differences of motor unit contractile properties in two most frequently studied mammals: cats and rats. A total sample of 166 motor units (79 in cats and 85 in rats) was investigated in the medial gastrocnemius muscle. Considerable differences were found in composition of the studied muscle. In cats, fast fatigable, fast resistant and slow units formed 68, 18 and 14% of the investigated population, whereas in rats 36, 52 and 12%, respectively. The contraction and relaxation times of motor units in the cat muscle were evidently longer than in the rat and the border values for fast/slow motor units division in these species were 44 and 20 ms, respectively. The mean values of twitch and tetanic forces appeared to be 7-8 times lower in rats, for fast, while 2-5 times for slow motor units. Also variability between the strongest and the weakest units within each type revealed differences 10-60 times in cats, whereas only 3.5-14 times in rats. The summation of twitches into tetanus for fast units was comparable in both species, but for S units was evidently more effective in the cat. In fast motor units' tetanic contractions evident interspecies differences concerned sag appearance and profiles of unfused tetani of FF and FR units. Differences in contractile properties described in the study may depend on the size, number and innervation ratio of motor units in the muscle of cat and rat, as well as their biochemical variability. Differences in composition of motor unit types and uneven mechanisms of force development may reflect biological adaptation to variable behaviour of cats and rats.     

Investigation of motor unit recruitment during stimulated contractions of tibialis anterior muscle

This work investigated motor unit (MU) recruitment during transcutaneous electrical stimulation (TES) of the tibialis anterior (TA) muscle, using experimental and simulated data. Surface electromyogram (EMG) and torque were measured during electrically-elicited contractions at different current intensities, on eight healthy subjects.EMG detected during stimulation (M-wave) was simulated selecting the elicited MUs on the basis of: (a) the simulated current density distribution in the territory of each MU and (b) the excitation threshold characteristic of the MU. Exerted force was simulated by adding the contribution of each of the elicited MUs. The effects of different fat layer thickness (between 2 and 8 mm), different distributions of excitation thresholds (random excitation threshold, higher threshold for larger MUs or smaller MUs), and different MU distributions within the muscle (random distribution, larger MU deeper in the muscle, smaller MU deeper) on EMG variables and torque were tested.Increase of the current intensity led to a first rapid increase of experimental M-wave amplitude, followed by a plateau. Further increases of the stimulation current determined an increase of the exerted force, without relevant changes of the M-wave. Similar results were obtained in simulations.Rate of change of conduction velocity (CV) and leading coefficient of the second order polynomial interpolating the force vs. stimulation level curve were estimated as a function of increasing current amplitudes. Experimental data showed an increase of estimated CV with increasing levels of the stimulation current (for all subjects) and a positive leading coefficient of force vs. stimulation current curve (for five of eight subjects). Simulations matched the experimental results only when larger MUs were preferably located deeper in the TA muscle (in line with a histochemical study). Marginal effect of MU excitation thresholds was observed, suggesting that MUs closer to the stimulation electrode are recruited first during TES regardless of their excitability.     

Recovery of rat tibialis anterior motor unit properties following partial denervation.

The recovery of selected mechanical, morphological, and metabolic properties of rat tibialis anterior fast motor units was determined following partial denervation (n = 7) or partial denervation and hemispinal cord transection (n = 5) and compared with age-matched control units (n = 7). Following 1-12 months of recovery, the mechanical properties of each unit were measured and the fibres depleted of glycogen by using standard ventral root filament stimulation techniques. Quantitative histochemical techniques were used to determine cross-sectional area and the activities of succinate dehydrogenase and alpha-glycerolphosphate dehydrogenase in individual unit fibres. Partial denervation increased the mean fibre area but decreased alpha-glycerolphosphate dehydrogenase activity. Succinate dehydrogenase was unchanged in the denervated groups. The variability in area and enzymatic activities among the unit fibres was unchanged. However, the interrelationship between the enzymes was altered by both denervation procedures. Succinate dehydrogenase activity was directly related to fatigue resistance and inversely related to tetanic tension across the units. These findings suggest that a motor unit reestablishes many of its properties despite marked changes to the composition of the unit brought about by partial denervation. In addition, a reduction in the neuromuscular activity of units during reorganization had a limited effect on recovery.     

Tetanic depression in fast motor units of the cat gastrocnemius muscle.

Ability of muscle fibers to generate force is decreased when higher frequency of stimulation of motor units immediately follows lower frequency. This phenomenon called tetanic depression was found in rat medial gastrocnemius. However, it was not clear whether tetanic depression occurred only in rat muscle or it concerns all mammals. This study was conducted on motor units of cat medial gastrocnemius. Analyses were made at three successive trains of stimulation: 30 Hz, 20 and 30 Hz and again 30 Hz (the first pattern) or 40 Hz, 25 and 40 Hz and 40 Hz (the second pattern). In all fast units force generated within the middle tetanus was lower than force generated at the same, but constant frequency of stimulation applied earlier or later. The mean tetanic depression in 30 Hz tetani amounted to 10.9% for fast fatigable (FF) and 15.9% for fast resistant (FR) motor units, whereas in 40 Hz tetani mean values were 5.6% and 7.3% for FF and FR motor units, respectively. In slow motor units tetanic depression was not observed. These results proved the existence of tetanic depression in the feline muscle and indicated that its intensity depends on the fusion of tetanus. It has been concluded, that the tetanic depression is a general property of fast motor units in mammals.     

Nonlinear summation of force in cat tibialis anterior: a muscle with intrafascicularly terminating fibers.

The complex connective tissue structure of muscle and tendon suggests that forces from two parts of a muscle may not summate linearly, particularly in muscles with intrafasciculary terminating fibers, such as cat tibialis anterior (TA). In four anesthetized cats, the TA was attached to a servomechanism to control muscle length and record force. The ventral roots were divided into two bundles, each innervating about half the TA, so the two parts could be stimulated alone or together. Nonlinear summation of force (F(nl)) was measured during isometric contractions. F(nl) was small and negative, indicating less than linear summation of the parts, which is consistent with the predicted F(nl) of muscle fibers connected in series. F(nl) was more significant when smaller parts of the muscle were tested (21.8 vs. 8% for whole muscle). These data were fit to a model where both parts of the muscle were assumed to stretch a common elasticity. Compensatory movements of the servomechanism showed the common elasticity is very stiff, and the model cannot account for F(nl) in cat TA.     

Force-velocity properties of fatigue-resistant units in cat fast-twitch muscle after fatigue

The isometric and force-velocity properties of an identified and uniform population of fast-twitch, fatigue-resistant (FR) fibers within the flexor digitorum longus (FDL) muscle were investigated before, immediately after, and during recovery from a fatiguing repetitive isometric stimulus regime (40 Hz for 330 ms every s for 180 s) in the anesthetized cat. It was necessary to determine the smallest fraction of muscle that had the same force-velocity properties as the whole muscle. This was approximately 15% for FDL; if the fraction was less, the maximum speed of shortening was depressed and the a/Po value increased. Motor units were enlarged by partial denervation of the muscle, causing the intact motoneurons to sprout and incorporate more muscle fibers; FR units showed the greatest increase. Immediately after the fatigue regime, maximum isometric tetanic tension declined to 67% but subsequently recovered to 90% of the control value by the end of the 60-min recovery period. Maximum speed of shortening dropped to 71% of the control but after 30 min had recovered and did not differ significantly from control values. It is concluded that the capacity for recovery from fatigue is greater for FR units than for a whole muscle, which also contains fast-fatiguable units, and that the mechanisms involved in the recovery of the maximum isometric tension and maximum speed of shortening are independently regulated.     

Associations between force and fatigue in fast-twitch motor units of a cat hindlimb muscle

Associations were quantified between the control force and fatigue-induced force decline in 22 single fast-twitch-fatigable motor units of 5 deeply anesthetized adult cats. The units were subjected to intermittent stimulation at 1 train/s for 360 s. Two stimulation patterns were delivered in a pseudo-random manner. The first was a 500-ms train with constant interpulse intervals. The second pattern had the same number of stimuli, mean stimulus rate, and stimulus duration, but the stimulus pulses were rearranged to increase the force produced by the units in the control (prefatigue) state. The associations among the control peak tetanic force of these units, 3 indices of fatigue, and total cumulative force during fatiguing contractions were dependent, in part, on the stimulation pattern used to produce fatigue. The associations were also dependent, albeit to a lesser extent, on the force measure (peak vs. integrated) and the fatigue index used to quantify fatigue. It is proposed that during high-force fatiguing contractions, neural mechanisms are potentially available to delay and reduce the fatigue of fast-twitch-fatigable units for brief, but functionally relevant, periods. In contrast, the fatigue of slow-twitch fatigue-resistant units seems more likely to be controlled largely, if not exclusively, by metabolic processes within their muscle cells.     

The motor units of cat medial gastrocnemius: electrical and mechanical properties as a function of muscle length.

The effects of changing muscle length on the mechanical properties of 89 motor units from adult cat medial gastrocnemius have been studied in eight experiments. Few differences were found between the effects of length on tetanic tension, twitch tension, twitch-tetanus ratio, twitch contraction time, twitch half relaxation time, rate of force development and electrical activity for fast contracting (twitch contraction time less than or equal to 45 msec) and slowly contracting (greater than 45 msec) units. Those differences that did appear did not persist when these two groups were matched by tetanic tension. It is concluded that the biophysical mechanisms responsible for the changes in mechanical and electrical properties with length must be similar for fast and slow twitch units and not related to potential differences in their muscle fiber type. The effects of changing muscle length on the mechanical properties of the eight whole muscles suggest that changes in force output with length are of minor importance during normal movements as the muscle is found to be electrically active over a relatively narrow range of lengths close to the optimum length for tetanus of the whole muscle. The very shortest muscle lengths at which there is only minimal force development are not used in natural movements, while the declining limb of the length tension curve is at muscle lengths beyond the maximum in situ length.     

Intramuscular pressure of human tibialis anterior muscle detects age-related changes in muscle performance

Intramuscular pressure (IMP) reflects forces produced by a muscle. Age is one of the determinants of skeletal muscle performance. The present study aimed to test whether IMP mirrors known age-related muscular changes. We simultaneously measured the tibialis anterior (TA) IMP, compound muscle action potential (CMAP), and ankle torque in thirteen older adults (60–80 years old) in vivo by applying different stimulation intensities and frequencies. We found significant positive correlations between the stimulation intensity and IMP and CMAP. Increasing stimulation frequency caused ankle torque and IMP to increase. The electromechanical delay (EMD) (36 ms) was longer than the onset of IMP (IMPD) (29 ms). Compared to the previously published data collected from young adults (21–40 years old) in identical conditions, the TA CMAP and IMP of older adults at maximum intensity of stimulation were 23.8% and 39.6% lower, respectively. For different stimulation frequencies, CMAP, IMP, as well as ankle torque of older adults were 20.5%, 24.2%, and 13.2% lower, respectively. Surprisingly, the EMD did not exhibit any difference between young and older adults and the IMPD was consistent with the EMD. Data supporting the hypotheses suggest that IMP measurement is an indicator of muscle performance in older adults.     

Further observations concerning the motor innervation of the tensor tympani muscle of the cat

The cat tensor tympani muscle presented an uncommon ultrastructural organization of neuromuscular junctions compared with those in the other striated muscles. In cross sections, individual neuromuscular junctions had very extended contact area of the nerve terminal and muscle fiber, the terminal bouton was covering as a "calyx" the postjunctional muscle fiber. Long basal lamina was interposed between them. The sarcolemma at the level of the nerve terminal had multiple infoldings along its length, or smooth postjunctional muscle membrane was found beneath endings on both fiber types.     

Heterogeneity of spindle units in the cat tenuissimus muscle

Three tandem spindles and their nerve supplies, reconstructed by light microscopy of serial transverse sections of the cat tenuissimus muscle, were compared to single spindle units. Each tandem spindle consisted of one large unit containing a dynamic bag1, a static bag2, and several static chain fibers (b1b2c unit) linked by the bag2 fiber to a small unit containing only a bag2 and chain fibers (b2c unit). Most features of primary afferents, secondary afferents, and motor neurons were qualitatively and quantitatively similar in both single and tandem b1b2c units. However, b1b2c units of tandem spindles had a lower density of skeletofusimotor innervation than did single b1b2c spindles. The b2c spindle units differed greatly from single or tandem b1b2c units. The b2c spindle units had fewer intrafusal fibers and incoming axons than either the tandem or single b1b2c units. The motor innervation of b2c units was typified by nonselective gamma axons that coinnervated both bag2 and chain fibers, in contrast to the regular occurrence of both selective and nonselective motor axons in b1b2c spindle units. The afferent located at the equator of b2c units differed in size, branching pattern, and intrafusal distribution of its ending from both the primary and secondary sensory axons of b1b2c units and, therefore, might represent a third category of spindle afferent. Thus, cat tenuissimus muscles contain three types of spindle units that differ in the number and organization of muscular and neural elements. These differences in structure and neural organization among tenuissimus spindle units may be a source for generation of different sensory signals in response to common mechanical or fusimotor stimuli.     

Proteomic analysis of rat soleus and tibialis anterior muscle following immobilization

A proteomic analysis was performed comparing normal slow twitch type fiber rat soleus muscle and normal fast twitch type fiber tibialis anterior muscle to immobilized soleus and tibialis anterior muscles at 0.5, 1, 2, 4, 6, 8 and 10 days post immobilization. Muscle mass measurements demonstrate mass changes throughout the period of immobilization. Proteomic analysis of normal and atrophied soleus muscle demonstrated statistically significant changes in the relative levels of 17 proteins. Proteomic analysis of normal and atrophied tibialis anterior muscle demonstrated statistically significant changes in the relative levels of 45 proteins. Protein identification using mass spectrometry was attempted for all differentially regulated proteins from both soleus and tibialis anterior muscles. Four differentially regulated soleus proteins and six differentially regulated tibialis anterior proteins were identified. The identified proteins can be grouped according to function as metabolic proteins, chaperone proteins, and contractile apparatus proteins. Together these data demonstrate that coordinated temporally regulated changes in the proteome occur during immobilization-induced atrophy in both slow twitch and fast twitch fiber type skeletal muscle.     

Damage to different motor units from active lengthening of the medial gastrocnemius muscle of the cat.

Slow-twitch motor units in the medial gastrocnemius muscle of the anesthetized cat were found to have an average optimum length for active tension that was 0.8 +/- 0.5 (SE) mm longer than the whole muscle optimum. For fast-twitch units (time to peak < 50 ms), the average optimum was 1.3 +/- 0.3 mm shorter than the whole muscle optimum. After the muscle had been subjected to 10 stretches while maximally activated, beginning at the whole muscle optimum length, the optimum lengths of the 27 fast-twitch motor units shifted significantly further in the direction of longer muscle lengths (mean 4.3 +/- 0.3 mm) than for the eight slow-twitch units (2.1 +/- 0.4 mm). A shift in the muscle's length-tension relation was interpreted as being due to sarcomere disruption. Statistical analysis showed that a motor unit's optimum length for a contraction, relative to the whole muscle optimum, was a better indicator of the unit's susceptibility to damage from active lengthenings than was motor unit type.     

Transcranial magnetic stimulation during resistance training of the tibialis anterior muscle.

During the first few weeks of resistance training, maximal voluntary contraction (MVC) force increases at a faster rate than can be accounted for by increases in protein synthesis. This early increase in MVC force has been attributed to neural mechanisms but the sources have not been identified. The purpose of this study was to measure changes in cortical excitability with transcranial magnetic stimulation during 4 weeks of resistance training of the tibialis anterior muscle. Ten individuals performed 6 sets of 10 MVCs 3 times per week for 4 weeks and ten participated as a control group. There were no changes in any parameters tested in the control group over the 4 weeks. In the training group, TA muscle strength increased significantly by 10% at week 2 and by 18% at week 4. As hypothesized, cortical excitability during resistance training also increased. The amplitude of the TA surface EMG motor evoked potential elicited by TMS during a low-level contraction increased by 32% after training with no change in the M-wave. These data indicate that there may be an increase in cortical excitability during the first few weeks of resistance training of the TA muscle.