3246671
Wheel-running exercise alters rat diaphragm action potentials and their regulation by K+ channels.
Endurance exercise modifies regulatory systems that control skeletal muscle Na+ and K+ fluxes, in particular Na+-K+-ATPase-mediated transport of these ions. Na+ and K+ ion channels also play important roles in the regulation of ionic movements, specifically mediating Na+ influx and K+ efflux that occur during contractions resulting from action potential depolarization and repolarization. Whether exercise alters skeletal muscle electrophysiological properties controlled by these ion channels is unclear. The present study tested the hypothesis that endurance exercise modifies diaphragm action potential properties. Exercised rats spent 8 wk with free access to running wheels, and they were compared with sedentary rats living in conventional rodent housing. Diaphragm muscle was subsequently removed under anesthesia and studied in vitro. Resting membrane potential was not affected by endurance exercise. Muscle from exercised rats had a slower rate of action potential repolarization than that of sedentary animals (P = 0.0098), whereas rate of depolarization was similar in the two groups. The K+ channel blocker 3,4-diaminopyridine slowed action potential repolarization and increased action potential area of both exercised and sedentary muscle. However, these effects were significantly smaller in diaphragm from exercised than sedentary rats. These data indicate that voluntary running slows diaphragm action potential repolarization, most likely by modulating K+ channel number or function.
25519681
Membrane potential and contractures in segments cut from rat fast and slow twitch muscles
Contractile responses associated with depolarization caused by an increase in [K]0 or by voltage-clamp steps were compared for fast and slow mammalian twitch muscles. The contractions and contractures of isolated mammalian muscle fibres cut into 0.5–1.0 cm lenghts are similar to those obtained from intact cells. The slope of the relationship between the membrane potential and the log [K]0 is similar in slow (46 mV±0.8) and in fast fibres (48 mV±1.1). This relation is not significantly modified in sodium-free or Cl-free solution. The K-contractures of cut sections of slow and fast fibres are transient and a full repriming of the response is only observed when the [K]·[Cl] product is kept constant. The contractile threshold in fast fibres is at 20–30 mM [K]0 (−52 to −43 mV) and in slow muscle at 10–15 mM [K]0 (−62 to −55 mV). Under voltage-clamp conditions, the contractile responses of both types of muscle show two components. In Na-free solution or in presence of TTX (5×10−7 g/ml) the first component is abolished and the second remaining component is similar to that developing during K-contractures. In iliacus fibres, the contracture threshold is between −50.5 mV and −40.5 mV and in soleus fibres between −66 mV and −56 mV; these values are close to those obtained with K-rich depolarizing fluids. The S-shaped curve of the contracture vs membrane potential relation is similar to that found in frog muscles except that the contractile responses are graded over a larger range of membrane potentials (−50 to + 30 mV in fast and −55 to +10 mV in slow muscle).
36422807
Use of ion channel blockers in studying the regulation of skeletal muscle contractions
SummaryEffects of K+- and Cl−-channel blockers on the muscle contraction of mouse diaphragm in response to direct electrical muscle stimulation were studied. K+-channel blockers (0.1–1 mmol/1 4-aminopyridine, 0.4–1.2 mmol/l uranyl nitrate and 2-30 mmol/l tetraethylammonium chloride) and a Cl−-channel blocker (0.01–0.03 mmol/1 9-anthracene carboxylic acid) increased the contractile amplitudes in a limited extent not to exceed over 50% of control. However, the sequential applications of two different channel blockers at a rather low concentration markedly increased the contractile responses mostly over 300% of control except the combination of 4-aminopyridine and uranyl nitrate. It appears that two K+-channel blockers synergistically exerted their effects rather than additionally in the regulation of muscle contractions. Investigation on the possible mechanism of the synergistic action of K+-channel blockers suggested that prolongation of action potential durations was in a linear correlation with the increased contractions. On the other hand, the contractile potentiation induced by combination of K+- and Cl−-channel blockers was attributed to the production of repetitive action potential firings (150±12 Hz) upon a single electrical stimulation. Similar to Cl−-channel blocker, low Cl− as well as low Ca2+ enchanced K+-channel blockers in producing contractile potentiation accompanied with stimulus-bound repetitive discharges. Tetrodotoxin at a concentration of 0.03 μmol/l which did not affect the twitches evoked by electrical stimulations completely inhibited the contractile potentiation induced by the combined application of K+- and Cl−-channel blockers. It was believed that these studies on the contractions of mouse diaphragm carried out in the physiological salt solution provided a better approach in exploring the possible functions of K+- and Cl−-channels in the regulation of skeletal muscle contractions. Moreover, because of the different K+-channels inhibited by these blockers (4-aminopyridine and uranyl nitrate majorly on delayed rectifier K+-channel and tetraethylammonium ion on Ca2+-activated and ATP-sensitive K+-channel), it was concluded that the different types of K+-channels as well as Cl−-channels exert their effects in a synergistic manner on the regulation of the skeletal muscle contractions.
44877881
Electrophysiologic and inotropic effects of K+-channel blockade in aged diaphragm.
The aminopyridines block several types of potassium (K+) channels and exert a direct inotropic effect on skeletal muscle by prolonging the duration of the action potential. Aging influences skeletal muscle Cl- channels and their regulation, and affects both resting whole-cell K+ conductance and adenosine triphosphate (ATP)-sensitive K+ channels, although in opposite directions. The present study tested the hypothesis that aging affects diaphragm-muscle K+ channels responsible for repolarization of the action potential and force production. Diaphragms of young adult (age 3 to 4 mo) and old (age 20 to 21 mo) male Fischer 344 rats was studied in vitro at 37 degrees C. The K+-channel blocker 3,4-diaminopyridine (DAP, 0.3 mM) did not alter resting membrane potential or action-potential height, overshoot, or rate of depolarization of either young-adult or old muscle. However, DAP slowed the rate of repolarization of the action potential and increased the action-potential area in young-adult and old muscle; the time for the action potential to repolarize by 80% increased from 0.59 +/- 0.02 ms (mean +/- SE) to 3.37 +/- 0.68 ms (p < 0.05) in young-adult muscle and from 0.87 +/- 0.06 ms to 2.52 +/- 0.54 ms (p < 0.05) in old muscle, whereas the action-potential area increased from 56 +/- 3 mVms to 193 +/- 34 mVms (p < 0.05) in young-adult muscle and from 72 +/- 5 mVms to 134 +/- 20 mVms (p < 0. 05) in old muscle. The action-potential area was not different in young-adult and old diaphragm without DAP, but was significantly larger in young-adult than in old diaphragm with DAP (p < 0.05). The functional consequence was that DAP increased diaphragm isometric twitch force by 181 +/- 12% (p < 0.05) in young-adult muscle and by 144 +/- 24% (p < 0.05) in old muscle; the increase was significantly greater in young-adult than in old muscle (p < 0.05). These data suggest an aging-associated reduction in, or reduced DAP sensitivity of, diaphragm K+ conductance during action potentials, which most likely reflects aging-associated alterations in delayed-rectifier K+ conductance. Although the inotropic effect of DAP was greater for young-adult than for old diaphragm muscle, the difference was sufficiently modest to show that DAP has substantial inotropic effects in old muscle.
1020240
Modulation of diaphragm action potentials by K(+) channel blockers.
K(+) channels regulate diaphragm contractility. The present study examined the electrophysiological mechanisms accounting for diversity among K(+) channel blockers in their inotropic actions on the diaphragm. Rat diaphragmatic muscle fibers were recorded intracellularly in vitro at 37 degrees C. Apamin and charybdotoxin (Ca2+)-activated K(+) channel blockers) did not alter resting membrane potential or action potentials. Glibenclamide (ATP-sensitive K(+) channel blocker) slowed action potential repolarization by 12% (P<0.05) and increased action potential area by 25% (P<0.005). Tetraethylammonium (which blocks several types of K(+) channels) increased action potential overshoot by 20% (P<0.01) and prolonged action potential rise time by 17% (P<0.02). 4-Aminopyridine and 3,4-diaminopyridine (which also block several types of K(+) channels) slowed action potential repolarization by 163% (P<0.0001) and 253% (P<0.0001), and increased action potential area by 183% (P<0.0001) and 298% (P<0.0001), respectively. Slowing of repolarization for the aminopyridines was especially marked at voltages approaching resting membrane potential, thereby changing action potential repolarization from a first to a second order decay. Previously reported variability in inotropic effects among K(+) channel blockers correlated significantly with the extent to which they slowed action potential repolarization and increased action potential area, but not with changes in other action potential properties.
96149557
Ionic channels of excitable membranes
PART I Classical biophysics of the squid giant axon The superfamily of voltage-gated channels Voltage-gated calcium channels Potassium channels and chloride channels Ligand-gated channels of fast chemical synapses Modulation, slow synaptic action, and second messengers Sensory transduction and excitable cells Calcium dynamics, epithelial transport, and intercellular coupling PART II Elementary properties of ions in solution Elementary properties of pores Counting channels Structure of channel proteins Selective permeability: Independence Selective permeability: Saturation and binding Classical mechanisms of block Structure-function studies of permeation and block Gating mechanisms: Kinetic thinking Gating: Voltage sensing and inactivation Modification of gating in voltage-sensitive channels Cell biology and channels Evolution and origins
24797569
Plasma potassium changes with high intensity exercise.
1. Exercise seems to change the extracellular potassium concentration far beyond the narrow limits seen in resting subjects. To examine alterations in plasma potassium concentration during exercise, twenty healthy, well‐trained men ran on the treadmill at 6 deg inclination with catheters inserted in the femoral vein and artery. 2. During 1 min exhausting exercise plasma potassium concentration rose in parallel in the vein and artery, reaching peak post‐exercise values of 8.34 +/‐ 0.23 mmol l‐1 and 8.17 +/‐ 0.29 mmol l‐1. After 3 min recovery the potassium concentration was 0.50 +/‐ 0.05 mmol l‐1 below pre‐exercise values. Both the rise of plasma potassium concentration during exercise and the decline during recovery followed exponential time courses with a half‐time of 25 s. 3. Exercise at reduced intensity showed that the peak post‐exercise potassium concentration was linearly related to the exercise intensity. Individual resting, peak and nadir values were proportionally related. 4. The increased potassium concentration during exercise can be explained in full by the electrical activity in the exercising muscles. Repeated 1 min exhausting exercise bouts revealed no relationship between potassium concentration and plasma pH nor glycogen break‐down. 5. All of the observations fit a simple model of potassium efflux from active muscle and elimination from blood with the following characteristics: the efflux increases (decreases) stepwise at the onset (end) of exercise, and the efflux rate during exercise increases with exercise intensity. Potassium is eliminated from blood by a proportional regulator which may be the Na(+)‐K+ pump of the exercising muscle. Extracellular potassium is indirectly linked to the pump stimulus, and the rate of reuptake is proportional to the extracellular accumulation. Thus no limited maximal power for potassium uptake was found. The post‐exercise undershoot of 0.5 mmol l‐1 can be explained by a higher gain of the pump after exercise. 6. The large, rapid changes in the plasma potassium concentration during and after exercise is due to the first order kinetics of the reuptake mechanism rather than to a limited power to take up potassium.
22717644
Role of potassium in the regulation of systemic physiological function during exercise.
In exercise, potassium (K+) is released from contracting muscle predominately through K+ channels associated with the repolarization phase of the action potential. Increases in extracellular K+ are directly related to increases in metabolic rate and may reach concentrations as high as 8-9 mM in the arterial blood during exhaustive work. Exercise-induced hyperkalaemia has been implicated in several physiological processes, in particular skeletal muscle fatigue, hyperaemia, pressor reflex, arterial chemosensitivity and myocardial stability. There is no direct evidence to show that hyperkalaemia causes muscle fatigue, although raised extracellular [K+] may contribute to fatigue during prolonged tetani by depressing the propagation of the action potential down the t-tubule system, thus impairing the release of Ca2+ from the sarcoplasmic reticulum. The vasodilating properties of K+ may transiently contribute to the early phase of exercise hyperaemia and interact synergistically with other vasoactive substances to cause relaxation by hyperpolarizing K+ channels in vascular smooth muscle. Hyperkalaemia has been implicated in the regulation of arterial blood pressure through activation of the muscle afferent reflex where potassium-depolarized C fibres may contribute to a reflex increase in arterial blood pressure. K+ can also increase ventilation and the sensitivity of the ventilatory response to hypoxia through direct excitation of the arterial chemoreceptors. Finally, to maintain myocardial electrical stability in exercise, there is a beneficial interaction between raised K+ and catecholamines on the heart, so that when they combine, each offsets the other's deleterious effects.
17590265
Dynamics and consequences of potassium shifts in skeletal muscle and heart during exercise.
Since it became clear that K(+) shifts with exercise are extensive and can cause more than a doubling of the extracellular [K(+)] ([K(+)](s)) as reviewed here, it has been suggested that these shifts may cause fatigue through the effect on muscle excitability and action potentials (AP). The cause of the K(+) shifts is a transient or long-lasting mismatch between outward repolarizing K(+) currents and K(+) influx carried by the Na(+)-K(+) pump. Several factors modify the effect of raised [K(+)](s) during exercise on membrane potential (E(m)) and force production. 1) Membrane conductance to K(+) is variable and controlled by various K(+) channels. Low relative K(+) conductance will reduce the contribution of [K(+)](s) to the E(m). In addition, high Cl(-) conductance may stabilize the E(m) during brief periods of large K(+) shifts. 2) The Na(+)-K(+) pump contributes with a hyperpolarizing current. 3) Cell swelling accompanies muscle contractions especially in fast-twitch muscle, although little in the heart. This will contribute considerably to the lowering of intracellular [K(+)] ([K(+)](c)) and will attenuate the exercise-induced rise of intracellular [Na(+)] ([Na(+)](c)). 4) The rise of [Na(+)](c) is sufficient to activate the Na(+)-K(+) pump to completely compensate increased K(+) release in the heart, yet not in skeletal muscle. In skeletal muscle there is strong evidence for control of pump activity not only through hormones, but through a hitherto unidentified mechanism. 5) Ionic shifts within the skeletal muscle t tubules and in the heart in extracellular clefts may markedly affect excitation-contraction coupling. 6) Age and state of training together with nutritional state modify muscle K(+) content and the abundance of Na(+)-K(+) pumps. We conclude that despite modifying factors coming into play during muscle activity, the K(+) shifts with high-intensity exercise may contribute substantially to fatigue in skeletal muscle, whereas in the heart, except during ischemia, the K(+) balance is controlled much more effectively.
188537
Exercise‐induced hyperkalaemia can be reduced in human subjects by moderate training without change in skeletal muscle Na, K‐ATPase concentration
Abstract. In 15 conscripts, venous plasma potassium was followed during exercise on a training bicycle before and after 10 weeks of moderate physical training and a putative relationship with skeletal muscle Na, K‐ATPase was evaluated. Peak plasma potassium concentration obtained at exhaustion was 61 pM 0.2 and 5.6 pM 0.2 mmol l‐1 (mean pM SEM, n= 14, P < 0.05) before and after training, respectively. Throughout the exercise period and within the first minutes of rest plasma potassium concentration was 0.2–0.5 mmol l‐1 higher before than after training. Neither peak values nor peak rises in plasma potassium concentration before nor after training were correlated to the 3H‐ouabain binding site (Na, K‐ATPase) concentration in vastus lateralis muscle. The results indicate that net loss of potassium from the skeletal muscle pool during exercise is reduced after training, that the heart during exercise may be exposed to a smaller rise in plasma potassium concentration after training than before, and that moderate improvement of capacity to clear extracellular potassium during exercise may be due to increased activity of existing Na, K‐pumps in resting skeletal muscle fibres. This may reduce muscle fatigue, increase physical performance and explain the paradoxical observation that, despite an increased catecholamine response, there is a reduced risk of cardiac events after training.
30857044
Muscle cell electrical hyperpolarization and reduced exercise hyperkalemia in physically conditioned dogs.
Contracting muscle cells release K ions into their surrounding interstitial fluid, and some of these ions, in turn, enter venous plasma. Thereby, intense or exhaustive exercise may result in hyperkalemia and potentially dangerous cardiotoxicity. Training not only reduces hyperkalemia produced by exercise but in addition, highly conditioned, long-distance runners may show resting hypokalemia that is not caused by K deficiency. To examine the factors underlying these changes, dogs were studied before and after 6 wk of training induced by running on the treadmill. Resting serum [K] fell from 4.2 +/- 0.2 to 3.9 +/- 0.3 meq/liter (P less than 0.001), muscle intracellular [K] rose from 139 +/- 7 to 148 +/- 14 meq/liter (P less than 0.001), and directly measured muscle cell membrane potential (Em) in vivo rose from -92 +/- 5 to -103 +/- 5 mV (P less than 0.001). Before training, resting Em of isolated intercostal muscle in vitro was -87 +/- 5 mV, and after incubation in 10(-4) M ouabain, Em fell to -78 +/- 5 mV. After training, resting Em of intercostal muscle rose to -95 +/- 4, but fell to -62 +/- 4 mV during incubation in 10(-4) M ouabain. The measured value for the Em was not completely explained by the increased ratio of intracellular to extracellular [K] or by the potassium diffusion potential. Skeletal muscle sarcolemmal Na,K-ATPase activity (microM inorganic phosphate mg-1 protein h-1) increased from 0.189 +/- 0.028 to 0.500 +/- 0.076 (P less than 0.05) after training, whereas activities of Mg2+ -dependent ATPase and 5'nucleotidase did not change. In untrained dogs, exercise to the point of exhaustion elevated serum [K] from 4.4 +/- 0.5 to 6.0 +/- 1.0 meq/liter (P less than 0.05). In trained dogs, exhaustive exercise was associated with elevation of serum [K] from 3.8 +/- 0.3 to 4.2 +/- 0.4 (NS). The different response of serum [K] to exercise after training was not explainable by blood pH. Basal insulin levels rose from 7.0 +/- 0.7 microU/ml in the untrained dogs to 9.9 +/- 1.0 microU/ml (P less than 0.05) after training. Although insulin might have played a role in the acquired electrical hyperpolarization, the reduced exercise-produced hyperkalemia after training was not reversed by blockade of insulin release with somatostatin. Although the fundamental mechanisms underlying the cellular hyperpolarization were not resolved, our observations suggest that increased Na-K exchange across the sarcolemmal membrane, the increase of Na,K-ATPase activity and possibly increased electrogenicity of the sodium pump may all play a role in the changes induced by training.
25497297
Sprint training increases human skeletal muscle Na(+)-K(+)-ATPase concentration and improves K+ regulation.
This study investigated the effects of sprint training on muscle Na(+)-K(+)-adenosinetriphosphatase (ATPase) concentration, plasma [K+] regulation, muscle performance, and fatigue during severe intermittent exercise. Six untrained male subjects underwent intensive cycle-sprint training for 7 wk. Muscle biopsies were taken at rest from the vastus lateralis muscle before and after 7 wk of training and were assayed for Na(+)-K(+)-ATPase concentration using vanadate-facilitated [3H]ouabain binding to intact samples. Before and after the training period, subjects performed four maximal 30-s exercise bouts (EB) on a cycle ergometer, each separated by a 4-min recovery. Arterialized venous blood samples were drawn immediately before and after each sprint bout and were analyzed for plasma [K+]. The work output was significantly elevated (11%) across all four EBs after training. The muscle [3H]ouabain binding site concentration was significantly increased (16%) from 333 +/- 19 to 387 +/- 15 (SE) pmol/g wet wt after training but was unchanged in muscle obtained from three control subjects. Plasma [K+] rose by 1-2 mmol/l with each EB and declined rapidly by the end of each recovery period. The increases in plasma [K+] resulting from each EB were significantly lower (19%) after training. The ratios of rise in plasma [K+] relative to work output during each EB were also significantly lower (27%) after training. The increased muscle [3H]ouabain binding site concentration and the reduced ratio of rise in [K+] relative to work output with exercise are both consistent with improved plasma and skeletal muscle K+ regulation after sprint training.
25062592
Training increases the concentration of [3H]ouabain-binding sites in rat skeletal muscle.
Exercise is associated with a net loss of K+ from the working muscles and an increased plasma K+ concentration, indicating that the capacity for intracellular reaccumulation of K+ is exceeded. Training reduces the exercise-induced rise in plasma K+, and an increased plasma [K+] may interfere with physical performance. Since the clearing of K+ from the extracellular space depends on the capacity for active K+ uptake in skeletal muscle, the effects of training and inactivity on the total concentration of (Na+ + K+)-ATPase was determined. Following 6 weeks of swim training, the concentration of [3H]ouabain-binding sites in rat hindlimb muscles was up to 46% (P less than 0.001) higher than in those obtained from age-matched controls. Whereas muscle Na+, K+ contents remained unchanged, the concentration of citrate synthase increased by up to 76% (P less than 0.001). Training induced no change in the [3H]ouabain-binding-site concentration in the diaphragm, but in the heart ventricles, the K+-dependent 3-O-methylfluorescein phosphatase activity increased by 20% (P less than 0.001). Muscle inactivity induced by denervation, plaster immobilisation or tenotomy reduced the [3H]ouabain-binding-site concentration by 20-30% (P less than 0.02-0.001) within 1 week. In conclusion, training leads to a significant and reversible rise in the concentration of (Na+ + K+)-ATPase in muscle cells. This may be of importance for the beneficial effects on physical performance by improving the maximum capacity for K+ clearance.
6406915
Habitual exercise enhances neuromuscular transmission efficacy of rat soleus muscle in situ.
Rat motor nerve terminals and the endplates they interact with exhibit changes to varying patterns of use, as when exposed to increased activation in the form of endurance exercise training. The extent to which these changes affect neuromuscular transmission efficacy is uncertain. In this study, the effects of habitual exercise on the electrophysiological properties of neuromuscular transmission in rat soleus muscle were investigated using a novel in situ approach. Consistent with previous reports, miniature endplate potential frequency was enhanced by habitual exercise. Other passive properties, such as resting membrane potential, miniature endplate potential amplitude, and "giant" miniature endplate potential characteristics were unaltered by the training program. Full-size endplate potentials were obtained by blocking soleus muscle action potentials with mu-conotoxin GIIIb. Quantal content values were 91.5 and 119.9 for control and active groups, respectively (P < 0.01). We also measured the rate and extent of endplate potential amplitude rundown during 3-s trains of continuous stimulation at 25, 50, and 75 Hz; at 50 and 75 Hz, we found both the rate and extent of rundown to be significantly attenuated (10--20%) in a specific population of cells from active rats (P < 0.05). The results establish the degree of activity-dependent plasticity as it pertains to neuromuscular transmission in a mammalian slow-twitch muscle.
6346504
Effects of DAP on diaphragm force and fatigue, including fatigue due to neurotransmission failure.
Among the aminopyridines, 3,4-diaminopyridine (DAP) is a more effective K+ channel blocker than is 4-aminopyridine (4-AP), and, furthermore, DAP enhances neuromuscular transmission. Because 4-AP improves muscle contractility, we hypothesized that DAP would also increase force and, in addition, ameliorate fatigue and improve the neurotransmission failure component of fatigue. Rat diaphragm strips were studied in vitro (37 degrees C). In field-stimulated muscle, 0.3 mM DAP significantly increased diaphragm twitch force, prolonged contraction time, and shifted the force-frequency relationship to the left without-altering peak tetanic force, resulting in increased force at stimulation frequencies < or = 50 Hz. During 20-Hz intermittent stimulation, DAP increased diaphragm peak force compared with control during a 150-s fatigue run and, furthermore, significantly improved maintenance of intratrain force. The relative contribution of neurotransmission failure to fatigue was estimated by comparing the force generated by phrenic nerve-stimulated muscles with that generated by curare-treated field-stimulated muscles. DAP significantly increased force in nerve-stimulated muscles and, in addition, reduced the neurotransmission failure contribution to diaphragm fatigue. Thus DAP increases muscle force at low-to-intermediate stimulation frequencies, improves overall force and intratrain fatigue during 20-Hz intermittent stimulation, and reduces neurotransmission failure.
34919331
Training-induced alterations in young and senescent rat diaphragm muscle.
The current study sought to examine the effects of chronic endurance treadmill running on oxidative capacity and capillary density in specific diaphragm muscle fiber types in young (5 mo) and senescent (greater than or equal to 23 mo) female Fischer 344 rats. Both young and senescent animals trained at approximately 75% of maximal O2 consumption for 1 h/day 5 days/wk for 10 wk. Plantaris citrate synthase activity was significantly increased (P less than 0.01) in both young and old trained groups. Densitometric analysis of succinate dehydrogenase (SDH) activity in diaphragm type I, IIa, and IIb muscle fibers was done using a computerized image-processing system. There were no age-related differences in SDH activity between the young and old groups for any of the fiber types. In addition, SDH activity was found to be significantly increased (P less than 0.05) in all three fiber types in both the young and senescent trained animals compared with their sedentary counterparts. Fiber size and capillary density did not differ between young and senescent rats, nor did exercise affect this measure. Each fiber, irrespective of type, had an average of approximately four capillaries in contact with it. However, type IIb fibers had a significantly lower capillary density per unit area than type I or IIa muscle fibers. The results indicate that the senescent costal diaphragm maintains its ability to adapt to an increased metabolic demand brought about by locomotor exercise. Of further interest is the finding that training adaptations occurred in all three fiber types, suggesting that increased work of breathing from moderate exercise leads to recruitment of all three fiber types.(ABSTRACT TRUNCATED AT 250 WORDS)
21278428
Exercise-induced cellular alterations in the diaphragm.
Limited data exist concerning the effects of exercise training on cellular oxidative capacity in the diaphragm of senescent animals. In this study we examined the changes in cellular oxidative capacity, muscle cell cross-sectional area (CSA), and capillarity within the costal diaphragm of senescent animals after a 10-wk endurance-training program. Twelve 24-mo-old female Fischer 344 rats were divided into either a sedentary control group (n = 6) or exercise training group (n = 6). The trained animals exercised on a motor-driven treadmill (60 min/day, 5 days/wk) at a work rate equal to approximately 55-65% VO2max. Capillaries were identified histologically and fiber types determined using adenosinetriphosphatase (ATPase) histochemistry. Succinate dehydrogenase (SDH) activity and CSA in individual fibers were measured using a computerized image analysis system. Exercise training did not increase (P > 0.05) the capillary-to-fiber ratio for any fiber type. However, training significantly decreased CSA (P < 0.05) and increased capillary density (capillary number/CSA) (P < 0.05) in type I, type IIa, and type IIb fibers. Furthermore, exercise training resulted in small but significant increase in SDH activity (P < 0.05) in type I and IIa fibers, whereas training did not alter SDH activity (P > 0.05) in type IIb fibers. These data demonstrate that endurance training in senescent animals results in small relative improvements in both oxidative capacity and capillary density in costal diaphragmatic type I and IIa muscle fibers. The increase in both capillary density and fiber SDH activity was largely due to a reduction in fiber CSA.
30865666
Aging and respiratory muscle metabolic plasticity: effects of endurance training.
We examined the oxidative and antioxidant enzyme activities in respiratory and locomotor muscles in response to endurance training in young and aging rats. Young adult (4-mo-old) and old (24-mo-old) female Fischer 344 rats were divided into four groups: 1) young trained (n = 12), 2) young untrained (n = 12), 3) old trained (n = 10), and 4) old untrained (n = 6). Both young and old endurance-trained animals performed the same training protocol during 10 wk of continuous treadmill exercise (60 min/day, 5 days/wk). Compared with young untrained animals, the young trained group had significantly elevated (P less than 0.05) activities of 3-hydroxyacyl-CoA dehydrogenase (HADH), glutathione peroxidase (GPX), and citrate synthase (CS) in both the costal diaphragm and the plantaris muscle. In contrast, training had no influence (P greater than 0.05) on the activity of lactate dehydrogenase within the costal diaphragm in young animals. In the aging animals, training did not alter (P greater than 0.05) activities of CS, HADH, GPX, or lactate dehydrogenase in the costal diaphragm but significantly (P less than 0.05) increased CS, HADH, and GPX activities in the plantaris muscle. Furthermore, training resulted in higher activities of CS and HADH in the intercostal muscles in the old trained than in the old untrained animals. Finally, activities of CS, HADH, and GPX were significantly (P less than 0.05) lower in the plantaris in the old untrained than in the young untrained animals; however, CS, HADH, and GPX activities were greater (P less than 0.05) in the costal diaphragm in the old sedentary than in the young untrained animals.(ABSTRACT TRUNCATED AT 250 WORDS)
21296156
Endurance training reduces the rate of diaphragm fatigue in vitro.
PURPOSE
The present study examined the effects of endurance training on the contractile and biochemical properties of the rat costal diaphragm in vitro.


METHODS
Sixty-four rats were divided into two groups: exercise trained (T) and control (C). Training consisted of treadmill running 5 d x wk(-1), 60 min x d(-1) at approximately 70% of VO2max, over a 10-wk period.


RESULTS
Control diaphragm strips produced an average of 12% less force from minute 15 to 50 of a 60-min in vitro fatigue protocol, compared with the T diaphragm strips (P < 0.01). T diaphragms had 10.1% higher citrate synthase (CS) and 12.1% higher superoxide dismutase (SOD) activities compared with the C (P < 0.05). Despite a significant decrease (P < 0.05) in Type IIb myosin heavy chains (MHC) and an increase (P < 0.05) in Type I MHC in T diaphragms, maximal shortening velocity (Vmax) in the diaphragm was not different between T and C animals. No differences were observed in specific force or the relative proportions of myosin light chains between groups.


CONCLUSIONS
These findings suggest that endurance training reduces the rate of diaphragm fatigue in vitro but has no effect on Vmax or specific force.
10140491
Impact of endurance concentric contraction training on acute force deficit following in vitro lengthening contractions
Abstract. The purposes of this study were to (1) determine whether endurance training employing solely concentric contractions would reduce force deficit following lengthening contractions, and (2) to determine if aged skeletal muscle would respond similarly from training, compared with young animals. Young (3-month) and old (23-month) male Fischer 344 rats were randomly assigned to either a control or an exercise training group. Exercise training consisted of 10 weeks of treadmill running (15% grade, 45 min/day, and 5 days/week) such that by the end of training the young and old rats were exercising at 27 and 15 m/min, respectively (~70% $$
\dot V{\rm O}_{{\rm 2max}} 
$$
). After training, the percent decrease in maximal isometric force (Po) of the diaphragm muscle was measured in vitro at 26°C immediately after a series of 20 lengthening contractions (20% strain from optimal muscle length, one contraction every 5 s), and at 5 and 10 min post-injury. At 10 min post-injury, Po averaged ~66% of initial Po in the exercise group (young and old combined) and ~61% of initial Po in the control groups (young and old combined) (P<0.05). This difference represents an attenuation of force deficit by ~13% in the trained animals, compared with controls. These findings indicate that 10 weeks of endurance training employing solely concentric contractions is beneficial in minimizing acute force deficit after a bout of lengthening contractions in both young and old rats.
606239
Treadmill running produces both positive and negative physiological adaptations in Sprague-Dawley rats.
Exercise training produces a vast array of physiological adaptations, ranging from changes in metabolism to muscle mitochondrial biogenesis. Researchers studying the physiological effects of exercise often use animal models that employ forced exercise regimens that include aversive motivation, which could activate the stress response. This study examined the effect of forced treadmill running (8 wk) on several physiological systems that are sensitive to training and stress. Forced treadmill running produced both positive and negative physiological adaptations. Indicative of positive training adaptations, exercised male Sprague-Dawley rats had a decrease in body weight gain and an increase in muscle citrate synthase activity compared with sedentary controls. In contrast, treadmill running also resulted in the potentially negative adaptations of adrenal hypertrophy, thymic involution, decreased serum corticosteroid binding globulin, elevated lymphocyte nitrite concentrations, suppressed lymphocyte proliferation, and suppressed antigen-specific IgM. Such alterations in neuroendocrine tissues and immune responses are commonly associated with chronic stress. Thus treadmill running produces both positive training adaptations and potentially negative adaptations that are indicative of chronic stress. Researchers employing forced activity need to be aware that this type of exercise procedure also produces physiological adaptations indicative of chronic stress and that these changes could potentially impact other measures of interest.
8633510
Developmental expression of voltage‐sensitive K+ channels in mouse skeletal muscle and C2C12 cells
The developmental expression of voltage‐sensitive K+ channels was analyzed by Northern blot in mouse skeletal muscle. Of nine Shaker‐like genes studied, eight are expressed in this mammalian muscle. Their expression is differentially regulated during development. The mouse cell line C2C12 has been used to study expression of voltage‐sensitive K+ channels during in vitro myotube differentiation. Different voltage‐sensitive K+ channel messages are also expressed in these cells which display a pattern of expression depending upon the differentiation stage. The message for the very peculiar K+ channel or IsK type could only be detected by polymerase chain reaction on skeletal muscle mRNA.
39074918
Expression of the potassium channel KV3.4 in mouse skeletal muscle parallels fiber type maturation and depends on excitation pattern
We report the detailed expression pattern of the voltage‐dependent potassium channel KV3.4 (rat homologue, Raw3) in mouse skeletal muscle. Using semi‐quantitative RT‐PCR, we show that its expression is detectable at embryonic day 17 and rises to adult levels within 2 weeks after birth. Expression is fiber type‐dependent, with mRNA levels being 5–6‐fold lower in the mixed slow/fast soleus muscle than in the fast tibialis anterior and extensor digitorum longus muscles. Fast muscles from myotonic mice exhibit low KV3.4 mRNA levels similar to those of wild‐type soleus. In denervated extensor digitorum longus, KV3.4 expression declines to perinatal levels. We conclude that KV3.4 expression in mouse skeletal muscle is regulated by the pattern of excitation.
40269827
Tolbutamide, but not glyburide, affects the excitability and contractility of unfatigued frog sartorius muscle.
The goal of this study was to characterize the effects of tolbutamide and glyburide, two known KATP channel blockers, on intact, unfatigued sartorius muscle fibres of the frog, Rana pipiens. Tetanic contractions were elicited by field stimulation with 200 ms long train of pulses (0.5 ms, 6 V, 140 Hz). Resting and action potentials were measured using conventional microelectrodes. At pHo 7.2 (extracellular pH), the tetanic force was unaffected by 0.5 mM and 1.0 mM tolbutamide, but at 2.0 mM it decreased by 15.5 +/- 1.0%. The effect of tolbutamide on the tetanic force was significantly greater at pHo 6.4: all three tolbutamide concentrations caused a significant decrease in tetanic force, being 62.3 +/- 9.4% at 2 mM. In the presence of tolbutamide a large number of fibres became unexcitable at pHo 6.4, but not at pHo 7.2. Glyburide at 10 microM, on the other hand, caused a 5-7% decrease in tetanic force at both pHo 6.4 and 7.2, but no further decreases in tetanic force were observed when the glyburide concentration was increased up to 100 microM. Unlike tolbutamide, glyburide did not affect the excitability of muscle fibres, but significantly prolonged the repolarization phase of action potentials, especially at pHo 6.4. We suggest that several of the tolbutamide effects reported in this study cannot be accounted for by a direct effect on KATP channels, and that the large decrease in membrane excitability and muscle contractility in the presence of tolbutamide must seriously be taken into consideration when this channel blocker is used to study the physiological role of KATP channels in intact muscle fibres.
26109190
Effect of exercise conditioning on excitation-contraction coupling in aged rats.
We studied aged (24-26 mo) Fischer 344 rats after they underwent 8 wk of moderate exercise conditioning. Right ventricular papillary muscles were loaded with the calcium indicator aequorin. Electrophysiological recordings were also performed. Time to peak isometric tension in muscles from exercised aged rats (EAR) was shorter than in those from unexercised aged rats (UAR) (126 +/- 7 vs. 167 +/- 7 ms; P less than 0.01). Time to 50% relaxation from peak isometric tension was also shorter in EAR than in UAR (88 +/- 3 vs. 119 +/- 12 ms; P less than 0.05). There was a trend toward decrease in time to peak light and a significant decrease in time to 50% decline from peak light (33 +/- 4 ms in EAR vs. 59 +/- 17 ms in UAR; P = 0.001). Action potential amplitude was smaller in EAR than in UAR (67 +/- 4 vs. 82 +/- 3 mV; P = 0.003); however, action potential duration was longer (137 +/- 6 ms in EAR vs. 100 +/- 10 ms in UAR; P = 0.005). Right ventricular-to-body weight ratios revealed no evidence of hypertrophy in EAR compared with UAR. Cardiac tissue norepinephrine content was significantly greater in EAR than in UAR (1,212 +/- 25 vs. 630 +/- 105 ng/tissue; P = 0.02). In summary, exercise reversed the age-related prolongation of isometric contraction and associated intracellular calcium transient in the aged rat while it prolonged the transmembrane action potential. In addition, exercise in aged rats resulted in an increase in cardiac norepinephrine content.
6725366
Endurance training alters outward K+ current characteristics in rat cardiocytes.
The effect of endurance run training on outward K+ currents with rapidly inactivating (I(to)) and sustained or slowly inactivating (I(sus)) characteristics was examined in left ventricular (LV) cardiocytes isolated from sedentary (Sed) and treadmill-trained (Tr) female Sprague-Dawley rats. Isolated LV cardiocytes were used in whole cell patch-clamp studies to characterize whole cell I(to) and I(sus). Peak I(to) was greatest in cells isolated from the Tr group. When I(to) was corrected for cell capacitance to yield a current density, most, but not all, of the Sed vs. Tr differences in I(to) magnitude were eliminated. Regardless of how I(to) was expressed (e.g., I(to) or I(to) density), the time required to achieve a peak value was markedly shortened in the cardiocytes isolated from the Tr group. Training elicited a reduction in I(sus) density. Action potential characteristics were determined in Sed and Tr cardiocytes in primary culture. Training did not affect resting membrane potential, whereas peak membrane potential was reduced and time to peak membrane potential was prolonged in the Tr group. In addition, time to 50% repolarization was significantly increased in cells from the Tr group. Collectively, these data indicate that I(to) and I(sus) characteristics are altered by training in isolated LV cardiocytes. These alterations in I(to) and I(sus) may be responsible, at least in part, for the training-induced alterations in action potential configuration in cardiocytes in primary culture.
24395709
Influence of exercise on excitation-contraction coupling in rat myocardium.
The present studies were conducted to investigate further the mechanisms by which the myocardium adapts to exercise training. Sixty female rats were randomly divided into sedentary control (group C) and trained (group T) groups. Group T was progressively trained for 12 wk. After the rats were killed, left ventricular papillary muscles were mounted in a tissue bath for mechanical studies. Muscles from group T generated greater peak isometric twitch tension per unit cross-sectional area than muscles from group C with [Ca2+]o ranging from 0.25 to 3.5 mM. Analyses of these data indicated that the Km for Ca2+ was not different but that the predicted number of sarcolemmal Ca2+ binding sites was 63% higher in group T. The ATPase activity of the purified cardiac myofibrils was not different between the two groups in the pCa range of 8.53-4.42. Action potentials were recorded with microelectrodes impaled into left ventricular muscle fibers of the subendocardium. Although there was no difference in the resting membrane potential, overshoot, or 90% duration, there was a significant prolongation of the action potential at 0 mV (20.2 +/- 1.0 vs 30.0 +/- 1.3 ms) in group T. These data further support the hypothesis that treadmill exercise enhances cardiac performance by increasing Ca2+ availability to the contractile element. This adaptation is mediated, at least in part, by a sarcolemmal adaptation induced by the exercise paradigm.
22839104
Voluntary exercise decreases progression of muscular dystrophy in diaphragm of mdx mice.
Effects of voluntary wheel running on contractile properties of diaphragm (DIA) and soleus (SOL) of dystrophic (mdx) and control (C57BL/10SNJ) mice were evaluated. In particular, we tested the hypothesis that daily voluntary running is not deleterious to muscle function in mdx mice. Both groups of mice ran extensively (control mice approximately 7 km/day, mdx mice approximately 5 km/day). Exercise increased maximal specific tetanus tension of mdx DIA from 1.02 +/- 0.04 to 1.33 +/- 0.06 kg/cm2 but did not restore it to the control level (2.55 +/- 0.17 kg/cm2). Maximal tetanus tension of sedentary mdx SOL (2.41 +/- 0.17 kg/cm2) was reduced compared with control (3.10 +/- 0.15 kg/cm2) and was not altered by running activity. Optimal length was significantly lower in DIA of mdx mice, and exercise did not change this. Fatigability and contractile properties of muscles measured in vitro were not altered by running activity with the exception of increased contraction time in mdx DIA. In conclusion, extensive wheel running is not deleterious to muscle function in mdx mice contrary to predictions of the "work overload" theory of muscular dystrophy. Rather, this exercise is beneficial for active tension generation of mdx DIA, the muscle most closely resembling muscles of patients with Duchenne muscular dystrophy.
34235747
Contractile and biochemical properties of diaphragm: effects of exercise training and fatigue.
The effect of high-intensity trained (6 X 4.5 min at 40 m/min, 15% grade, 2.5-min rest between bouts, 5 days/wk, for 6 wk) on contractile, biochemical, and fatigue properties of the rat diaphragm were examined. The exercise program produced significant elevations in the mitochondrial marker enzyme citrate synthase (mumol X g-1 X min-1) in the soleus (SOL) (27.2 +/- 1.5 vs. 46.7 +/- 2.4; mean +/- SE), deep vastus lateralis (DVL) (40.8 +/- 2.6 vs. 58.3 +/- 2.8), and superficial vastus lateralis (SVL) (8.5 +/- 0.6 vs. 11.4 +/- 0.7). No significant differences were observed in the crural (CRU) (45.9 +/- 2.0 vs. 44.0 +/- 2.3) or ventral costal (VEN) (41.5 +/- 2.0 vs. 45.8 +/- 2.6) diaphragmatic regions. Phosphofructokinase, the rate-limiting enzyme of glycolysis, significantly increased in the SOL (19.0 +/- 0.8 vs. 23.3 +/- 1.3 mumol X g-1 X min-1) and DVL (69.3 +/- 6.0 vs. 86.6 +/- 5.0), but no alterations were seen in the SVL (98.6 +/- 5.7 vs. 106.1 +/- 9.0), CRU (54.4 +/- 2.8 vs. 53.8 +/- 1.5), or VEN (44.7 +/- 2.4 vs. 46.4 +/- 1.4) posttraining. Diaphragm contractile properties, with the exception of an increased rate of fall in twitch tension, remained unchanged after training. Glycogen values were significantly higher in trained diaphragms at rest (6.54 +/- 0.39 vs. 4.86 +/- 0.41 mg/g) and during 1, 5, and 10 min of fatiguing stimulation. During fatigue no differences were observed in force, rate of rise in force, rate of fall in force, muscle lactate, ATP, or creatine phosphate in trained vs. control.(ABSTRACT TRUNCATED AT 250 WORDS)
26731672
Role of sarcolemma action potentials and excitability in muscle fatigue.
The purposes of this study were to characterize the alterations in the sarcolemma action potential (AP) waveform and sarcolemma excitability as a result of fatiguing stimulation of the frog semitendinosus muscle and to relate these changes to the decrease in the force-generating ability of the muscle. Trains of APs were recorded before and after stimulation (100-ms trains, 150 Hz, 1/s for 5 min). The resting membrane potential (RMP), AP overshoot (OS), and duration at 50% of peak magnitude (DUR) were -84.3 +/- 2.0 mV, 19.5 +/- 1.9 mV, and 1.3 +/- 0.1 ms, respectively, before stimulation. The stimulation protocol caused RMP to depolarize to -75.1 +/- 2.0 mV, OS to fall to 7.3 +/- 1.9 mV, and DUR to increase to 2.5 +/- 0.4 ms. RMP and OS recovered fully in 5 min after the cessation of stimulation, whereas DUR was still prolonged. Before the stimulation protocol, AP frequency matched the stimulation frequency at all stimulation rates < or = 150 Hz. At 200-Hz stimulation, AP frequency was 192 +/- 6 Hz. After 5 min of stimulation, AP frequency matched the stimulation frequency only at < or = 60 Hz. At 100-, 150-, and 200-Hz stimulation, AP frequencies were 89 +/- 8, 84 +/- 17, and 79 +/- 15 Hz, respectively. Because of a decreased fusion frequency at fatigue, the fall in the sarcolemma AP frequency did not contribute to the decreased force. The stimulation-induced alterations in the AP waveform were moderate and unlikely to have caused fatigue. However, the alterations in AP may have been more extreme in the depths of the transverse tubules.