Double-sigmoid model for fitting fatigue profiles in mouse fast- and slow-twitch muscle

We present a curve-fitting approach that permits quantitative comparisons of fatigue profiles obtained with different stimulation protocols in isolated slow-twitch soleus and fast-twitch extensor digitorum longus (EDL) muscles of mice. Profiles from our usual stimulation protocol (125 Hz for 500 ms, evoked once every second for 100-300 s) could be fitted by single-term functions (sigmoids or exponentials) but not by a double exponential. A clearly superior fit, as confirmed by the Akaiki Information Criterion, was achieved using a double-sigmoid function. Fitting accuracy was exceptional; mean square errors were typically <1% and r(2) > 0.9995. The first sigmoid (early fatigue) involved approximately 10% decline of isometric force to an intermediate plateau in both muscle types; the second sigmoid (late fatigue) involved a reduction of force to a final plateau, the decline being 83% of initial force in EDL and 63% of initial force in soleus. The maximal slope of each sigmoid was seven- to eightfold greater in EDL than in soleus. The general applicability of the model was tested by fitting profiles with a severe force loss arising from repeated tetanic stimulation evoked at different frequencies or rest periods, or with excitation via nerve terminals in soleus. Late fatigue, which was absent at 30 Hz, occurred earlier and to a greater extent at 125 than 50 Hz. The model captured small changes in rate of late fatigue for nerve terminal versus sarcolemmal stimulation. We conclude that a double-sigmoid expression is a useful and accurate model to characterize fatigue in isolated muscle preparations.     

Junctional transmission in fast- and slow-twitch mammalian motor units

The phenomena of facilitation and tetanic potentiation of end-plate potentials (EPPs) were investigated in the predominantly fast-twitch extensor digitorum longus (EDL) and the predominantly slow-twitch soleus (Sol) muscles of the rat, using extra- and intracellular recording methods. When the quantal content of the EPPs was reduced to average values of 2-3, facilitation and potentiation could be studied in both muscles without the masking effects of neuromuscular depression. Under these conditions, the facilitation and potentiation at the neuromuscular junctions of EDL were different from those at the junctions of Sol. The potentiation during sustained activation was higher in EDL than in Sol at stimulation frequencies varying between 10 and 100 Hz. The same tendency was found for the facilitation measured by paired stimuli; the facilitation in EDL was generally higher than in Sol with interpulse intervals between 300 and 10 ms. The dependence of facilitation and potentiation on the stimulation frequency was also different in EDL and Sol. As the interpulse interval decreased from 200 to 25 ms, the facilitation in Sol increased more steeply than in EDL, whereas with further decrease from 25 to 10 ms, the facilitation in Sol did not increase at all, whereas the facilitation in EDL exhibited a substantial increase. The same general pattern was observed for potentiation: the potentiation in Sol increased more steeply than in EDL between 10 and 40 Hz (100- to 25-ms intervals) and more moderately than in EDL between 40 and 100 Hz (25- to 10-ms intervals). Another paradigm, repetitive short trains, induced a progressive increase in potentiation at the neuromuscular junctions of both muscles. The cumulative effect of potentiation was more prominent in EDL than in Sol. The differences in potentiation between EDL and Sol were also evident from intracellular recordings. These recordings also revealed that the amplitude of miniature EPPs remained unaltered in both muscles before and during the stimulus trains. Comparisons of the EPPs elicited before and at the end of tetanic trains revealed that the onset time of the EPPs was delayed in Sol but not in EDL during the train. This latency shift was also observed in the focally recorded nerve terminal potentials and end-plate currents in Sol, but not in EDL. The differences in facilitation and potentiation between EDL and Sol may be due to heterogeneities of the transmitter release machinery of their nerve terminals or differences in the capability of their presynaptic nerves to conduct action potentials at high frequencies.(ABSTRACT TRUNCATED AT 400 WORDS)     

Effects of adrenaline on excitation-induced stimulation of the sodium-potassium pump in rat skeletal muscle

Experiments were performed on isolated rat soleus and extensor digitorum longus (EDL) muscles of 4-week-old rats. In the soleus, direct electrical stimulation for 10 min induced a frequency-dependent increase in the ouabain-suppressible 86Rb+ uptake, which was maximal (+110%) at a frequency of 2 Hz. In the EDL this frequency only induced a 31% increase. A supramaximal concentration of adrenaline (10 mumol l-1) stimulated ouabain-suppressible 86Rb+ uptake by 80% and 27% in soleus and EDL, respectively. The combined effect of stimulation at 2 Hz and adrenaline was not significantly larger than each of the interventions alone in either of the muscles. The fractional loss of 22Na+ from soleus muscle was increased by around 50% by the exposure to adrenaline, electrical stimulation at 2 Hz or a combination of both. The effect of electrical stimulation on 22Na+ efflux was not prevented by addition of propranolol (1 or 10 mumol l-1). The results indicate that the stimulation of active Na+-K+ transport induced by adrenaline or electrical stimulation is much more pronounced in soleus (slow-twitch) muscle than in EDL (fast-twitch) muscle. Since it has been suggested that an accumulation of K+ ions in the extracellular space may play a role in the development of fatigue (Bigland-Ritchie 1984), our findings might be related to the fact that slow-twitch muscles have a much higher resistance to fatigue than fast-twitch muscles (Burke et al. 1971).     

The Effects of Dietary Creatine Supplements on the Contractile Properties of Rat Soleus and Extensor Digitorum Longus Muscles

Daily creatine supplements (0.258 g kg(-1) ) were administered to adult male Wistar rats (n = 7) in the drinking water. Age matched rats (n = 6) acted as controls. After 5-6 days, contractile properties were examined in soleus and extensor digitorum longus (EDL) muscle strips in vitro at 30 degrees C. In soleus muscles, creatine supplements decreased the half-relaxation time of the isometric twitch from 53.6 +/- 4.3 ms in control muscles to 48.4 +/- 5.5 ms but had no effect on twitch or tetanic tension or on twitch contraction time. In EDL muscles twitch tension, tetanic tension, twitch contraction and half-relaxation times were all unaffected by creatine supplements. Creatine supplements increased the fatigue resistance of the soleus muscles but had no effect on that of the EDL muscles. After a 5 min low-frequency fatigue test, tension (expressed as a percentage of initial tension) was 56 +/- 3 % in control soleus muscles, whereas that in the creatine-supplemented muscles was 78 +/- 6 % (P < 0.01). In the EDL muscles, the corresponding values were 40 +/- 2 % and 41 +/- 9 %, respectively. The force potentiation which occurred in the EDL muscles during the initial 20-30 s of the fatigue test was 170 +/- 10 % of initial tension in the control muscles 24 s after the initial stimulus train but was reduced (P < 0.01) to 130 +/- 20 % in the creatine-supplemented muscles. In conclusion, soleus muscle endurance was increased by creatine supplements. EDL endurance was unaffected but force potentiation during repetitive stimulation was decreased. Experimental Physiology (2001) 86.2, 185-190.     

The influence of secretin on the secretion of pepsin in response to acid stimulants in the anaesthetized cat.

1. The problem of selectivity during reinnervation of skeletal muscle fibres was investigated in the rat using the fast-twitch extensor digitorum longus (EDL) and the slow-twitch soleus muscles and their nerves. 2. After an operation on these nerves permitting them to compete for reinnervation of one or the other muscle (hereafter called Y-union), virtually the total isometric tetanic tension of EDL muscle could be elicited by stimulating the EDL nerve, while stimulating the soleus nerve yielded little or no tension. In the case of the soleus muscle, stimulation of either nerve elicited about half of the total isometric tetanic tension. 3. During the course of reinnervation of these muscles in non-competitive situations, the time course of increase in the ratio of tension elicited by nerve stimulation to that by direct stimulation was slower in the case of soleus nerve reinnervating EDL muscle, compared with cross-reinnervation in the reverse direction or reinnervation of each muscle by its own nerve. 4. Crushing the common peroneal nerve 12 days after a Y-union in an attempt to retard the EDL nerve did not favour reinnervation of the EDL by soleus nerve, but crushing the nerve again or just once at 1 month after the original operation produced substantial partial reinnervation of the EDL by the soleus nerve. 5. It is concluded that soleus nerve fibres form functioning neuromuscular synapses on EDL muscle fibres only with difficulty. The pattern of reinnervation reveals characteristic differences between fast-twitch and slow-twitch muscles on the one hand and between their respective nerves on the other.     

Muscle performance following fatigue induced by isotonic and quasi-isometric contractions of rat extensor digitorum longus and soleus muscles in vitro

AIM: To study the effect of contraction mode on fatigue development. METHODS: Muscle fatigue was induced by isotonic and quasi-isometric contractions in rat soleus (SOL) and extensor digitorum longus (EDL) muscles, using identical stimulation protocol (60 Hz, 400 ms s-1) for 100 s in SOL and 60 s in EDL. Fatigue was quantified as the decline in peak values of shortening, shortening velocity, relaxation and work during the isotonic contractions, and, correspondingly, of force, rate of force development, relaxation and work during the quasi-isometric contractions. Maximal test contractions (60 Hz, 1.5 s) performed before and after fatigue were analysed for decline in force development (Fmax), rate of force development (dF/dtmax) and relaxation (-dF/dtmax). RESULTS: Fmax declined to significantly lower values after isotonic than after quasi-isometric fatiguing contractions (fatigued in percentage of unfatigued): 58.5 +/- 6.4% vs. 64.4 +/- 7.0% in SOL, and 30.4 +/- 4.1% vs. 33.3 +/- 3.6% in EDL, respectively. The same pattern was seen for dF/dtmax which decreased to: 46.3 +/- 9.9% vs. 52.3 +/- 8.5% in SOL, and 19.1 +/- 4.3% vs. 22.3 +/- 3.2% in EDL after isotonic and quasi-isometric contractions, respectively. Similarly, when comparing fatigue development during the two contraction modes, the respective fatigue variables decreased more rapidly and to lower levels during isotonic vs. quasi-isometric contractions. During maximal test contractions, the dynamic fatigue variables (+/-dF/dtmax) declined to significantly lower levels than Fmax. CONCLUSIONS: Fatigue development was significantly larger during isotonic vs. quasi-isometric contractions. The use of force as the only experimental fatigue variable may underestimate the functional impairment of fatigued muscle, neglecting the fatigue effect on time and length dimensions.     

Adaptation of rat extensor digitorum longus to overload and increased activity

Rat extensor digitorum longus (EDL) muscles were overloaded by removal of the synergist tibialis anterior (TA). The weight of the overloaded muscle was increased 15 days after the initial operation and remained higher throughout the period studied (153 days). The times to peak twitch tension and half relaxation remained unaltered, but the twitch and tetanic tensions developed by the overloaded EDL muscles increased. The overloaded EDL muscles became significantly more fatigue resistant. In a separate group of animals the overloaded EDL muscle was also chronically stimulated at 10 Hz. The additional stimulation altered the response of the EDL to overload in that the time to peak twitch tension of the muscle was slightly prolonged. There was no increase in twitch or tetanic tension in spite of the increase in muscle weight, but the electrical stimulation led to a further increase in fatigue resistance above that seen in overloaded muscles. The histochemical and immunocytochemical examination of the muscle revealed that there was a moderate increase in succinate dehydrogenase activity in the muscles overloaded only, but a considerable increase in those overloaded muscles that were also stimulated. There was no obvious change in the number of muscle fibres that reacted with an antibody to slow myosin in either overloaded only or overloaded and stimulated EDL muscles. Thus the addition of continuous activity to overload induced a slowing of contraction and prevented the increase of force usually induced by overload.     

Slow skeletal muscles of the mouse have greater initial efficiency than fast muscles but the same net efficiency

The aim of this study was to determine whether the net efficiency of mammalian muscles depends on muscle fibre type. Experiments were performed in vitro (35°C) using bundles of muscle fibres from the slow-twitch soleus and fast-twitch extensor digitorum longus (EDL) muscles of the mouse. The contraction protocol consisted of 10 brief contractions, with a cyclic length change in each contraction cycle. Work output and heat production were measured and enthalpy output (work + heat) was used as the index of energy expenditure. Initial efficiency was defined as the ratio of work output to enthalpy output during the first 1 s of activity. Net efficiency was defined as the ratio of the total work produced in all the contractions to the total, suprabasal enthalpy produced in response to the contraction series, i.e. net efficiency incorporates both initial and recovery metabolism. Initial efficiency was greater in soleus (30 ± 1%; n = 6) than EDL (23 ± 1%; n = 6) but there was no difference in net efficiency between the two muscles (12.6 ± 0.7% for soleus and 11.7 ± 0.5% for EDL). Therefore, more recovery heat was produced per unit of initial energy expenditure in soleus than EDL. The calculated efficiency of oxidative phosphorylation was lower in soleus than EDL. The difference in recovery metabolism between soleus and EDL is unlikely to be due to effects of changes in intracellular pH on the enthalpy change associated with PCr hydrolysis. It is suggested that the functionally important specialization of slow-twitch muscle is its low rate of energy use rather than high efficiency.     

Activation of glycogen phosphorylase by electrical stimulation of isolated fast-twitch and slow-twitch muscles from rat

The influence of muscle contraction, induced by electrical stimulation, on the activity of glycogen phosphorylase, the contents of high-energy phosphates, hexose-monophosphates and lactate have been studied in isolated extensor digitorum longus (EDL) and soleus muscles from rats. The activity of phosphorylase a + b was about nine times higher in fast twitch muscles (EDL) than in slow-twitch soleus and remained unchanged during the stimulation. A pronounced increase of phosphorylase a occurred during the stimulation in EDL muscle. Stimulation with a frequency of 50 Hz for 10 s and 2 Hz for 90 s resulted in a 44-fold and five-fold increase in phosphorylase a, respectively. In contrast, stimulation of soleus muscle resulted in only a minor increase of phosphorylase a. The rate of glycogenolysis increased in both muscles during the stimulation but the increase was four to five times higher in the EDL than in soleus muscle. The content of phosphocreatine (PCr) before stimulation was much higher in EDL than in soleus but similar after the stimulation. This resulted in a three- to four-fold higher release of inorganic phosphate (Pi) in EDL than in soleus during contraction. Pi has previously been shown to be present in a limiting amount for the activity of phosphorylase and the increase during contraction is of importance for increasing the glycogenolytic rate. It is concluded that the higher glycogenolytic capacity in fast-twitch muscles compared to slow-twitch muscles is due to: (1) higher content of phosphorylase a + b, (2) higher degree of transformation of the enzyme into the a form during contraction, and (3) higher content of PCr, which liberates a large amount of Pi during contraction.     

Age-dependent fatigue in single intact fast- and slow fibers from mouse EDL and soleus skeletal muscles

In the present work, we investigate age-dependent changes in isometric endurance in response to repetitive stimulation in single intact fast- and slow-twitch muscle fibers from young and old mice. To examine this issue we performed in vitro experiments in manually dissected EDL and soleus muscle fibers. We examined the force generation capacity of fibers in response to two stimulation protocols characterized by different inter-tetanic intervals, named short (1-s) and long interval (3.65-s). Fatigability was measured according to the fatigue index (FI, ratio between the maximum tension recorded in the last over the first tetanus in a train of pulses), the time course of the FI and sag (gradual decrease in force during a partially fused tetanic contraction). Fibers were classified according to the FI using two different criteria previously used in the literature (first criterion: FI > or = 1, 075-099, 0.5-074 and < 0.5; second criterion: FI > or = 1, 0.75-0.99, 0.25-0.74 and < 0.25). The fatigue index distribution recorded in the population of fibers corresponding to EDL and soleus muscles from young and old mice studied with the short and long interval protocols was not statistically different. In summary, these results support the concept that the decline in mechanical performance with aging is not related with changes in fatigability of individual fast- or slow twitch muscles fibers.     

Velocity recovery cycles of C fibres innervating human skin

The purpose of this study was to determine whether induced expression of the Ca²⁺ buffering protein parvalbumin (PV) in slow-twitch fibres would lead to alterations in physiological, biochemical and molecular properties reflective of a fast fibre phenotype. Transgenic (TG) mice were generated that overexpressed PV in slow (type I) muscle fibres. In soleus muscle (SOL; 58 % type I fibres) total PV expression was 2- to 6-fold higher in TG compared to wild-type (WT) mice. Maximum twitch and tetanic tensions were similar in WT and TG but force at subtetanic frequencies (30 and 50 Hz) was reduced in TG SOL. Twitch time-to-peak tension and half-relaxation time were significantly decreased in TG SOL (time-to-peak tension: 39.3 ± 2.6 vs. 55.1 ± 4.7 ms; half-relaxation time: 42.1 ± 3.5 vs. 68.1 ± 9.6 ms,   P < 0.05  for TG vs. WT, respectively;   n = 8  –10). There was a significant increase in expression of type IIa myosin heavy chain (MHC) and ryanodine receptor at the mRNA level in TG SOL but there were no differences in MHC expression at the protein level and thus no difference in fibre type. Whole muscle succinate dehydrogenase activity was reduced by 12 ± 0.4 % in TG SOL and single fibre glycerol-3-phosphate dehydrogenase activity was decreased in a subset of type IIa fibres. These differences were associated with a 64 % reduction in calcineurin activity in TG SOL. These data show that overexpression of PV, resulting in decreased calcineurin activity, can alter the functional and metabolic profile of muscle and influence the expression of key marker genes in a predominantly slow-twitch muscle with minimal effects on the expression of muscle contractile proteins.     

Functional regeneration in the hindlimb skeletal muscle of the mdx mouse

Summary The pattern of spontaneous skeletal muscle degeneration and clinical recovery in hindlimb muscles of the mdx mutant mouse was examined for functional and metabolic confirmation of apparent structural regeneration. The contractile properties, histochemical staining and myosin light chain and parvalbumin contents of extensor digitorum longus (EDL) and soleus (Sol) muscles of mdx and age-matched control mice were studied at 3–4 and 32 weeks. Histochemical staining (myofibrillar ATPase and NADH-tetrazolium reductase) revealed no significant change in slow-twitch-oxidative (SO) or fast-twitch-oxidative-glycolytic (FOG) fibre type proportions in mdx Sol apart from the normal age-related increase in SO fibres. At 32 weeks mdx EDL, however, showed significantly smaller fast-twitch-glycolytic (FG) and larger FOG proportions than those in control EDL. These fibre type distributions were confirmed by differential staining with antibodies to myosin slow-twitch and fast-twitch heavy chain isozymes. Frequency distribution of cross-sectional area for each fibre type showed a wider than normal range of areas especially in FOG fibres of mdx Sol, and FG fibres of mdx EDL, supporting previous observations using autoradiography of myofibre regeneration. Isometric twitch and tetanic tensions in Sol were significantly less than in controls at 4 weeks, but by 32 weeks, values were not different from age-matched controls. In mdx EDL at 3 weeks, twitch and tetanus tensions were significantly less, and time-to-peak twitch tensions were significantly faster than in control EDL. By 32 weeks, mdx EDL twitch and tetanus tensions expressed relative to muscle weight continued to be significantly lower than in age-matched controls, despite normal absolute tensions. The maximum velocity of shortening in 32-week mdx EDL was significantly lower than in control EDL. Myosin light chain distribution in mdx Sol exhibited significantly less light chain 2-slow (LC2s) and more light chain 1b-slow(LC1bs) at 32 weeks than age-matched control Sol. Gels of EDL from 32-week-old mdx mice showed significantly less light chain 2-fast-phosphorylated (LC2f-P) and light chain 3-fast (LC3f) and significantly more light chain 1-fast (LC1f) and light chain 2-fast (LC2f), but normal parvalbumin content compared to age-matched controls. These observations suggest that mdx hindlimb muscles are differentially affected by the disease process as it occurs in murine models of dystrophy. However, the uniqueness of mdx Sol and to a lesser extent EDL is that they also undergo an important degree of functional regeneration which is able to compensate spontaneously for degenerative influences of genetic origin. The mdx mutant may therefore be an important model for the study of regeneration by skeletal muscle, and of the nerve-muscle interactions which enable or restrict that regeneration.     

Disuse of rat muscle in vivo reduces protein kinase C activity controlling the sarcolemma chloride conductance

Muscle disuse produced by hindlimb unloading (HU) induces severe atrophy and slow-to-fast fibre type transition of the slow-twitch soleus muscle (Sol). After 2 weeks HU, the resting ClC-1 chloride conductance ( g _Cl) of sarcolemma, which controls muscle excitability, increases in Sol toward a value typical of the fast-twitch EDL muscle. After 3 days of HU, the g _Cl increases as well before initiation of fibre type transition. Since ClC-1 channels are acutely silenced by PKC-dependent phosphorylation, we studied the modulation of g _Cl by PKC and serine–threonine phosphatase in Sol during HU, using a number of pharmacological tools. We show that a fraction of ClC-1 channels of control Sol are maintained in an inactive state by PKC basal activity, which contributes to the lower g _Cl in control Sol compared to EDL. After 14 days of HU, PKC/phosphatase manipulation produces effects on Sol g _Cl that corroborate the partial slow-to-fast transition. After 3 days of HU, the early increase of g _Cl in Sol is entirely attributable to a reduction of PKC activity and/or activation of phosphatase, maintaining ClC-1 channels in a fully active state. Accordingly, we found that HU reduces expression of PKCα, ɛ, and θ isoenzymes in Sol and EDL muscles and reduces total PKC activity. Moreover, we show that the rheobase current is increased in Sol muscle fibres as soon as after 3 days of HU, most probably in relation to the increased g _Cl. In conclusion, Sol muscle disuse is characterized by a rapid reduction of PKC activity, which reduces muscle excitability and is likely to contribute to disuse-induced muscle impairment.     

Skeletal muscle adaptation to physical training and beta-adrenergic blockade in spontaneously hypertensive rats

Summary The effects of training alone or in combination with long-term, non-selective, -adrenergic blockade on histochemical and biochemical properties of fast-twitch [extensor digitorum longus muscle (EDL)] and slow-twitch [soleus muscle (Sol)] muscle were analyzed in spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto strain rats (WKY). Fiber type distribution of Sol was drastically modified in SHR with fewer type I fibers and more type IIA fibers. No such histochemical alterations were observed in EDL. While prolonged swimming training remained ineffective in inducing both histochemical and biochemical improvement in WKY, SHR displayed a significant enhancement of capillarization and oxidative capacity in both Sol and EDL. However, in long-term -blocks rats training failed to improve significantly the oxidative capacity of SHR muscles, suggesting that -adrenoreceptor stimulation is necessary for a fully efficient adaptation of muscular metabolism to physical training.     

The influence of catecholamines on Na,K transport in slow- and fast-twitch muscles of the rat

The effects of catecholamines on active sodium and potassium transport was compared in slow- (SOL) and fast-twitch (EDL) skeletal muscles of the rat. Stimulation of active Na+-extrusion and K+-uptake induced by adrenaline (6-30 mumol . l-1) and isoprenaline (1-40 mumol . l-1) was markedly greater in slow- than in fast-twitch muscle. In sodium-preloaded muscles the maximal stimulation of 24Na-efflux induced by adrenaline was about 2-fold higher in SOL than in EDL. Isoprenaline caused a 2.4-fold increase in ouabain-sensitive 86Rb influx in SOL muscle, but failed to alter the ouabain-sensitive influx in EDL. The stimulating action of isoprenaline on 86Rb influx in EDL was due to an increase in the ouabain-insensitive fraction of Rb uptake. The effects of catecholamines of fast- and slow-twitch muscles were probably due to the accumulation of cyclic AMP, however the fact that there were no significant differences between the nucleotides levels in fast- and slow-twitch muscle suggests the participation of other mechanism as well. The results presented suggest that cyclic AMP-induced stimulation of ouabain-insensitive transport of cation in the isolated EDL muscle of the rat is similar to that of barnacle muscle.     

Muscle performance following fatigue induced by isotonic and quasi‐isometric contractions of rat extensor digitorum longus and soleus muscles in vitro

Aim: To study the effect of contraction mode on fatigue development. Methods: Muscle fatigue was induced by isotonic and quasi‐isometric contractions in rat soleus (SOL) and extensor digitorum longus (EDL) muscles, using identical stimulation protocol (60 Hz, 400 ms s^?1) for 100 s in SOL and 60 s in EDL. Fatigue was quantified as the decline in peak values of shortening, shortening velocity, relaxation and work during the isotonic contractions, and, correspondingly, of force, rate of force development, relaxation and work during the quasi‐isometric contractions. Maximal test contractions (60 Hz, 1.5 s) performed before and after fatigue were analysed for decline in force development (F_max), rate of force development (dF/dt_max) and relaxation (?dF/dt_max). Results: F _max declined to significantly lower values after isotonic than after quasi‐isometric fatiguing contractions (fatigued in percentage of unfatigued): 58.5 ± 6.4% vs. 64.4 ± 7.0% in SOL, and 30.4 ± 4.1% vs. 33.3 ± 3.6% in EDL, respectively. The same pattern was seen for dF/dt_max which decreased to: 46.3 ± 9.9% vs. 52.3 ± 8.5% in SOL, and 19.1 ± 4.3% vs. 22.3 ± 3.2% in EDL after isotonic and quasi‐isometric contractions, respectively. Similarly, when comparing fatigue development during the two contraction modes, the respective fatigue variables decreased more rapidly and to lower levels during isotonic vs. quasi‐isometric contractions. During maximal test contractions, the dynamic fatigue variables (±dF/dt_max) declined to significantly lower levels than F_max. Conclusions: Fatigue development was significantly larger during isotonic vs. quasi‐isometric contractions. The use of force as the only experimental fatigue variable may underestimate the functional impairment of fatigued muscle, neglecting the fatigue effect on time and length dimensions.     

Limited oxygen diffusion accelerates fatigue development in mouse skeletal muscle

Isolated whole skeletal muscles fatigue more rapidly than isolated single muscle fibres. We have now employed this difference to study mechanisms of skeletal muscle fatigue. Isolated whole soleus and extensor digitorum longus (EDL) muscles were fatigued by repeated tetanic stimulation while measuring force production. Neither application of 10 m m lactic acid nor increasing the [K⁺] of the bath solution from 5 to 10 m m had any significant effect on the rate of force decline during fatigue induced by repeated brief tetani. Soleus muscles fatigued slightly faster during continuous tetanic stimulation in 10 m m [K⁺]. Inhibition of mitochondrial respiration with cyanide resulted in a faster fatigue development in both soleus and EDL muscles. Single soleus muscle fibres were fatigued by repeated tetani while measuring force and myoplasmic free [Ca²⁺] ([Ca²⁺]_i). Under control conditions, the single fibres were substantially more fatigue resistant than the whole soleus muscles; tetanic force at the end of a series of 100 tetani was reduced by about 10% and 50%, respectively. However, in the presence of cyanide, fatigue developed at a similar rate in whole muscles and single fibres, and tetanic force at the end of fatiguing stimulation was reduced by ∼80%. The force decrease in the presence of cyanide was associated with a ∼50% decrease in tetanic [Ca²⁺]_i, compared with an increase of ∼20% without cyanide. In conclusion, lactic acid or [K⁺] has little impact on fatigue induced by repeated tetani, whereas hypoxia speeds up fatigue development and this is mainly due to an impaired Ca²⁺ release from the sarcoplasmic reticulum.     

Properties of motor units in nerve-intact autografts of cat extensor digitorum longus muscles

1. In cats, isometric contractile properties were measured on five extensor digitorum longus (EDL) muscles and four EDL muscle grafts 150-270 days after autografting with nerves intact. Comparisons were made between the properties of whole muscles and grafts and between 36 motor units in control EDL muscles and 41 motor units in grafts. 2. The time-to-peak twitch force (TPT) of 23 +/- 1.7 (SE) ms for grafts was significantly prolonged compared with the value of 17 +/- 0.7 ms observed for whole EDL muscles. The mean values for the TPT of motor units were not different from the respective values for whole grafts or for whole muscles. The maximum specific force of whole grafts of 19.7 +/- 0.6 (SE) N/cm2 was significantly less than the control value of 23.6 +/- 0.6 N/cm2, an observation consistent with all previous data on the maximum specific force of grafts and control muscles. 3. Based on the presence or absence of sag and an index of fatigue, motor units were classified as fast fatigable (FF), fast intermediate (FI), fast fatigue-resistant (FR), and slow (S). Motor units were classified 33% FF, 22% FI, 27% FR, and 17% S in control muscles and 17% FF, 43% FI, 29% FR, and 12% S in autografted muscles. Compared with control muscles, the number of small FF units increased significantly in the autografts, but no significant difference was observed in the fatigue properties of motor units.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of long-term cold exposure on contractile muscles of rats

The effects of 20-week cold exposure on contractile properties of soleus and extensor digitorum longus (EDL) muscles and plasma hormone levels were studied in rats. Twenty male Wistar rats (5 week old) were randomly divided into 2 groups (n = 10 each): cage-control and cold-exposed. The rats in the cold-exposed group were immersed in shoulder-deep water (approximately 18 degrees C) for 1 h/d, 5 d/week, for 20 weeks. The temperature and humidity of the animal room with 12:12 h light-dark cycle were maintained at approximately 23 degrees C and 55%, respectively. The rats were pair-fed powdered diets. The electromyogram activities in soleus and EDL were elevated by cold exposure. The body weight and absolute soleus wet weight of the cold-exposed group were significantly less than controls at the end of experiment. The one-half relaxation time and contraction time of EDL were significantly longer in the cold-exposed group than in the control group. The rate of twitch tension development, normalized by the maximum twitch tension, in EDL of the cold-exposed group was less than in the control group. Further, the fatigue resistance of EDL, but not of soleus, in response to train stimulation at 10 Hz was improved by cold exposure. The plasma levels of thyroid hormones, 3,5,3'-triiodothyronine and thyroxine, were significantly greater in cold-exposed group. Similar changes were also seen in the plasma catecholamine levels in the cold-exposed group (p > 0.05). It is suggested that long-term cold exposure causes a shift of the contractile properties of fast-twitch EDL muscle toward the slow-twitch type. The results also indicated that the characteristics of muscles responded more strongly to an increased activity level than to the elevation of plasma hormones.