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Influence of microtubules on vascular smooth muscle contraction.

Microtubules are ubiquitous in eukaryotic cells and play key roles in many cellular activities. The purpose of this study was to investigate the influence of microtubules on vascular smooth muscle contraction. Quantitative immunocytochemical analysis of rat aortic tissue revealed that, relative to the control group, colchicine (15 muM, 90 min) and nocodazole (15 muM, 90 min) decreased the microtubule density by 40-50% while taxol (10 muM, 90 min) increased the microtubule density by 33%. Isometric contraction studies demonstrated that both colchicine and nocodazole caused an upward shift in the phenylephrine (10(-8) to 10(-5) M) dose-response curve while taxol caused no significant change when compared to the control group. Potassium chloride (30 mM) induced 55 +/- 5% P0 contraction in DMSO treated vessel rings. The active tension increased to 73 +/- 5% P0 and 71 +/- 6% P0 after pretreatment of the aortic rings with colchicine or nocodazole, respectively. Taxol did not cause a significant change in the active tension (56 +/- 7% P0). These results indicate that microtubule depolymerization enhances isometric contraction of vascular smooth muscle and this enhanced contraction is not receptor dependent. Pretreatment of the aortic rings with an inhibitor of nitric oxide synthase (NOS) (Nomega-nitro-L-arginine) did not change the increased contractile response to phenylephrine due to microtubule depolymerization suggesting that this phenomenon is not mediated by endothelium dependent relaxation.

Animals↗

Activation varies among the knee extensor muscles during a submaximal fatiguing contraction in the seated and supine postures.

Ten young men sustained an isometric contraction of the knee extensor muscles at 20% of the maximum voluntary contraction (MVC) torque on three separate occasions in a seated posture. Subjects performed an isometric knee extension contraction on a fourth occasion in a supine posture. The time to task failure for the seated posture was similar across sessions (291 +/- 84 s; P > 0.05), and the MVC torque was similarly reduced across sessions after the fatiguing contraction (42 +/- 12%). The rate of increase in electromyograph (EMG) activity (%MVC) and torque fluctuations during the fatiguing contractions were similar across sessions. However, the rate of increase in EMG differed among the knee extensor muscles: the rectus femoris began at a greater amplitude (31.5 +/- 11.0%) compared with the vastus lateralis and vastus medialis muscles (18.8 +/- 5.3%), but it ended at a similar value (45.4 +/- 3.1%). The time to task failure and increase in EMG activity were similar for the seated and supine tasks; however, the reduction in MVC torque was greater for the seated posture. These findings indicate that the time to task failure for the knee extensor muscles that have a common tendon insertion did not alter over repeat sessions as had been observed for the elbow flexor muscles (Hunter SK and Enoka RM. J Appl Physiol 94: 108-118, 2003).

Adult↗

Acute and sustained effects of isometric and lengthening muscle contractions on high-energy phosphates and glycogen metabolism in rat tibialis anterior muscle.

Previous studies have shown that lengthening contractions, in contrast to isometric contractions, readily result in sustained malfunctioning of the exercised muscles. The present study was performed to investigate whether an exercise period with many (240) lengthening contractions (LC) results in alterations in muscle high-energy phosphates and inosine monophosphate (IMP) content, different from muscles performing a few (60) lengthening or a few (60) or many (240) isometric contractions (IC). Moreover, we sought for a possible cause(s) of the inability to replenish muscle glycogen stores following LC. Rat tibialis anterior muscles were subjected in vivo to either 60 or 240 LC or IC. Structural muscle damage occurred only after 240 LC. The fact that tissue glycogen levels declined to a similar extent during LC and IC suggests that the energy demand was comparable during both types of exercise. Nevertheless, the observation that on the one hand tissue stores of adenine nucleotides showed a greater decline, and on the other hand the tissue content of IMP increased to a significantly higher level after LC than after IC, clearly indicates that muscle energy metabolism is more disturbed during LC than during IC. The high tissue levels of IMP may contribute to impaired mechanical function as previously observed in muscles subjected to LC. In contrast to 240 IC, 24 hours after 240 LC, tissue glycogen stores and high-energy phosphate levels were not restored to control values. The present findings indicate that depressed glycogen synthase activity and impaired activity of the mitochondrial marker enzyme cytochrome C oxidase probably contribute to a continuous disturbance of energy metabolism in the exercised muscles during the 24 hours following 240 LC.

Adenosine Diphosphate↗

[Role of K+, Na+, and Ca2+ ions in the cardiodepressive action of blood plasma in burn shock and the crush syndrome].

Ion-selective electrodes were employed to measure the concentration of K+, Na+ and Ca2+ in blood plasma of rabbits with burn shock or crush syndrome (CS). No significant changes in the plasma concentration of Na+, and Ca2+ were found under both pathological conditions. The plasma concentration of K+ in burn shock significantly increased from 3.06 +/- 0.73 (control) to 5.28 +/- 2.65 mM (n = 10), whereas in CS from 3.42 +/- 1.03 to 4.92 +/- 1,29 mM (n = 8). The rise of K+ concentration in the control plasma to the maximal values seen in the "burn" and "syndrome" plasma led to an increase in the duration of intracellular action potentials (AP) but did not substantially change the amplitude of isometric contractions of the papillary muscles of rabbit heart. Meanwhile the similar rise of the duration of intracellular AP during perfusion of the papillary muscles with the "burn" and "syndrome" plasma was accompanied by an appreciable drop of the amplitude of isometric contractions. It is suggested that elevation of K+ concentration in blood plasma, inducing an increase in the duration of intracellular AP of cardiocytes may be responsible for changes in the ECG in burn and CS. At the same time inhibition of myocardial contractility in burn shock and CS is virtually not linked with hyperkalemia.

Animals↗

The energy cost of relaxation in control and hypertrophic rabbit papillary muscles.

The energy cost of the onset and relaxation phases of cardiac isometric contractions has been investigated by ergometer controlled-length releases occurring at different times during the contraction cycle, to test the hypothesis that the energy cost of relaxation is normally small. Energy flux has been measured myothermically in 20 or 30 contractions of rabbit papillary muscles. The ergometer releases took place after different delays, starting during the latency period and incrementing in 50 ms steps, until eventually, releases were occurring late into the relaxation phase. The release step was kept constant and of a magnitude sufficient to prevent significant redevelopment of active stress at any release interval. The rate of release was several times greater than the maximum shortening velocity of the papillary muscle preparations. The heat production in each train of contractions was measured, but in order to estimate the total energy output, the elastic energy in the muscle-lever system which was removed by the ergometer release had to be added to the heat. This was estimated by integration of the stress-strain relationship found for each muscle. In normal animals the contraction peak, at 27 degrees C and a 1.0 Hz stimulus rate, was located between the 215 and 265 ms release times, at which point the total energy flux was estimated to be 80%-90% of that measured in a normal isometric contraction. Additional experiments were performed in a group of volume-overloaded hearts and the data were compared with results from sham-operated controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Recovery of isometric grip strength after Colles' fracture: a prospective two-year study.

Grip strength during short and sustained maximal voluntary isometric contractions was measured in 28 females and 5 males with displaced Colles' fracture involving the distal radio-ulnar joint. The patients were randomized into two groups, treated either through immobilization with plaster cast or with external fixation. The recovery of isometric grip strength was followed over a two-year period. A significant difference was registered between women with plaster casts and women with external fixators six weeks after the fracture. Regaining of grip strength occurred up to one year after the fracture. The pattern of recovery was slower for women with primary external fixation. Neither the dominant nor the non-dominant injured side regained short or sustained maximal voluntary isometric contraction. The dominant injured side showed no significant difference between sides but the non-dominant injured side remained significantly weaker. It is thus important to identify hand dominance. Pain during measurements was reduced after two years, but about one-fifth of the patients still perceived pain. The present findings may serve as guidance in physiotherapy for these patients.

Aged↗

Time-resolved X-ray diffraction studies of myosin head movements in live frog sartorius muscle during isometric and isotonic contractions.

Using the facilities at the Daresbury Synchrotron Radiation Source, meridional diffraction patterns of muscles at ca 8 degrees C were recorded with a time resolution of 2 or 4 ms. In isometric contractions tetanic peak tension (P0) is reached in ca 400 ms. Under such conditions, following stimulation from rest, the timing of changes in the major reflections (the 38.2 nm troponin reflection, and the 21.5 and 14.34/14.58 nm myosin reflections) can be explained in terms of four types of time courses: K1, K2, K3 and K4. The onset of K1 occurs immediately after stimulation, but that of K2, K3 and K4 is delayed by a latent period of ca 16 ms. Relative to the end of their own latent periods the half-times for K1, K2, K3 and K4 are 14-16, 16, 32 and 52 ms, respectively. In half-times, K1, K2, K3 lead tension rise by 52, 36 and 20 ms, respectively. K4 parallels the time course of tension rise. From an analysis of the data we conclude that K1 reflects thin filament activation which involves the troponin system; K2 arises from an order-disorder transition during which the register between the filaments is lost; K3 is due to the formation of an acto-myosin complex which (at P0) causes 70% or more of the heads to diffract with actin-based periodicities; and K4 is caused by a change in the axial orientation of the myosin heads (relative to thin filament axis) which is estimated to be from 65-70 degrees at rest to ca 90 degrees at P0. Isotonic contraction experiments showed that during shortening under a load of ca 0.27 P0, at least 85% of the heads (relative to those forming an acto-myosin complex at P0) diffract with actin-based periodicities, whilst their axial orientation does not change from that at rest. During shortening under a negligible load, at most 5-10% of the heads (relative to those forming an acto-myosin complex at P0) diffract with actin-based periodicities, and their axial orientation also remains the same as that at rest. This suggests that in isometric contractions the change in axial orientation is not the cause of active tension production, but rather the result of it. Analysis of the data reveals that independent of load, the extent of asynchronous axial motions executed by most of the cycling heads is no more than 0.5-0.65 nm greater than at rest.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Effects of neuromuscular-blocking drugs in rats in vivo: direct measurements in the diaphragm and tibialis anterior muscle.

BACKGROUND: Effects of neuromuscular-blocking drugs are often investigated in rodents using both the diaphragm in vitro and the tibialis anterior muscle in vivo. Differences in the properties of these muscles cannot be neglected. We evaluated the neuromuscular effects of drugs on the rat diaphragm in vivo in comparison with the tibialis anterior muscle, directly measuring twitch tension with a force transducer. METHODS: Respectively using sternotomy and an approach via the gluteal space, transducers were attached to the diaphragm and the tibialis anterior muscle. Phrenic and sciatic nerves were stimulated supramaximally by single square pulses to record isometric contraction of muscles. Pancuronium, vecuronium, or rocuronium was infused continuously at a rate adjusted repeatedly until stable tension was obtained. Effective doses for 50% (ED 50) and 90% (ED 90) depression of tension were calculated, using a cumulative infusion dosing method. RESULTS: Isometric contraction of the diaphragm could be recorded despite movements of the heart and lungs. For all three drugs, both mean ED 50 and ED 90 were more than twice as large for the diaphragm as for the tibialis anterior (P<0.05). In addition, slopes of dose-response curves for the diaphragm were significantly less steep than those of the tibialis anterior for the three drugs. CONCLUSION: Neuromuscular-blocking effects on the diaphragm can be recorded in vivo. According to direct contractile force measurement, the diaphragm is more resistant to non-depolarizing neuromuscular-blocking drugs than the tibialis anterior muscle.

Animals↗

Estimates of cellular mechanics in an arterial smooth muscle.

Estimates of force generation or shortening obtained from smooth muscle tissues are valid for individual cells only if each cell is contracting homogeneously and if cells anatomically arranged in series are mechanically coupled. These two assumptions were tested and shown to be valid for the pig carotid media under certain conditions. Homogeneity of cellular responses in carotid strips was estimated from the motion of markers on the tissue during K+ -induced isometric contractions. When tissues were stretched to L0 (the optimum length for force generation), there was little marker movement on stimulation. However, considerable marker movement was observed on stimulation at shorter muscle lengths, reflecting localized shortening or stretching. The mechanical coupling of the very small cells in the media was determined by measuring the dependence of cell length on tissue length. Tissues were fixed with glutaraldehyde during isometric contractions at various tissue lengths (0.4--1.1 x L0). The fixed tissues were macerated with acid and the lengths of the dispersed cells were measured. Cell lengths were broadly distributed at all muscle lengths. However, the direct proportionality between mean cell length and muscle length (as a fraction of L0) indicated that cells which are anatomically in series are coupled force-transmitting structures. We conclude that valid estimates of cellular mechanical function in this preparation can be obtained from tissue measurements at lengths greater than about 0.9L0.

Animals↗

Central versus peripheral adaptations following eccentric resistance training.

Aim of the present investigation was to study the effects of an eccentric training on the neuromuscular properties of the plantar-flexor muscles. The experiment was carried out on 14 males divided into two groups (eccentric and control). Eccentric training consisted of six sets of six eccentric contractions at 120 % of one maximal concentric repetition and it was performed four times a week during four weeks. Before and after the 4-wk period, the plantar-flexor torque and the associated electromyographic activity were recorded during voluntary contractions (isometric, concentric and eccentric) and electrically induced contractions (twitch and tetanus), in order to distinguish central from peripheral adaptations. For the eccentric group, voluntary torque significantly increased after training independent of the action mode (relative gains 14 - 30 %, p < 0.05). This was associated with an increase in agonist EMG activity during isometric action and a decrease in antagonist coactivation in concentric (-27 %) and eccentric actions (-22 %) (p < 0.05). Voluntary activation level significantly increased from 80 +/- 5 % to 91 +/- 2 % (p < 0.05). Some of the twitch contractile properties (peak torque and maximal rate of twitch tension relaxation) were significantly modified (p < 0.05), but no changes were observed for the tetanus characteristics. These results allowed to conclude that the torque gains observed after the present training were more likely associated to central adaptations, affecting both agonist and antagonist muscles.

Adaptation, Physiological↗

The role of prostaglandins in the spontaneous motility of the fallopian tube.

Spontaneous isometric contractions of the human fallopian tube were measured in vitro. The tubes were obtained during the follicular phase of the menstrual cycle in 16 experiments and during the luteal phase in 23 experiments. The area under the curve and the frequency of spontaneous isometric contractions, together with the amount of prostaglandins E and F produced by the tube, were measured. Using each tissue as its own control, the effects of adding indomethacin or papeverine to the tissue both were assessed. Indomethacin, a prostaglandin synthetase inhibitor, significantly reduced the prostaglandin output without affecting the tubal motility. By contrast, papaverine, which is a smooth muscle relaxant but not an inhibitor of prostaglandin synthesis, significantly reduced both motility and prostaglandin output. It is concluded that prostaglandins do not have a direct role in the regulation of the spontaneous motility of the human fallopian tube.

Fallopian Tubes↗

The inotropic effects of a synthetic pyrethroid decamethrin on isolated guinea pig atrial muscle.

Pyrethroid insecticides have been reported to increase transmembrane sodium influx and inhibit ion-dependent ATPases in insect, squid, and toad nerve tissues. Since changes in sarcolemmal ion fluxes and inhibition of membrane-bound ATPases can alter myocardial contractility, the effects of a potent synthetic pyrethroid, decamethrin, on mammalian myocardium were examined using isolated left atrial muscle of the guinea-pig heart electrically stimulated at 1.5 Hz. Decamethrin (0.1-10 microM) increased the force of isometric contraction in a dose-dependent manner without affecting the resting tension. Propranolol (5 microM) reduced the magnitude of the pyrethroid's inotropic effect; however, a significant inotropic effect was still observed with 1 and 10 microM decamethrin in the presence of this concentration of propranolol. Similar results were noted in the presence of 500 microM procainamide and in left atrial muscle obtained from reserpine-treated animals. The results suggest that decamethrin increased the force of myocardial contraction by two mechanisms; indirectly by releasing catecholamines from sympathetic nerve terminals and directly by an action on the myocardium. Inotropic concentrations of the pyrethroid did not inhibit partially purified rat brain or guinea pig heart Na+, K+-ATPase. Moreover, twitch tension recordings of isometric contractions showed that, in the presence of propranolol, decamethrin increased tension development without changing either the time to peak tension or the duration of contraction. Tetrodotoxin almost completely abolished the positive inotropic effect of decamethrin. Thus, the positive inotropic effect of decamethrin is apparently due to an increase in transmembrane sodium influx which causes catecholamine release from the sympathetic nerve terminals and also directly enhances muscle contraction.

Animals↗

Estrogen protects against hypertension in the spontaneously hypertensive rat, but its protective mechanism is unrelated to impaired arterial muscle relaxation.

OBJECTIVES: To determine if estrogen acutely and directly alters arterial muscle relaxation, if estrogen is responsible for gender dichotomy in hypertension, and if arterial muscle from female spontaneously hypertensive rats (SHR) is slow to relax as is muscle from male SHR compared with arterial muscle of normotensive Wistar-Kyoto rats (WKY). METHODS: Relaxation rates of isometrically contracted arterial muscle from male rats were measured before and after addition of beta-estradiol. Blood pressure (BP) was monitored in intact male and female SHR and WKY and in ovariectomized and estrone-treated ovariectomized SHR and WKY. Relaxation rates of maximum isometric contractions of arterial muscle excised from male SHR and WKY and female SHR and WKY with varying chronic estrogen status were measured. RESULTS: Beta-estradiol had no direct, acute effect on arterial muscle force or relaxation. Intact and estrone-implanted SHR females had significantly lower BP than males. Ovariectomized SHR developed high BP equivalent to that of males. Arterial relaxation was slower in both male and female SHR compared with WKY. CONCLUSIONS: Estrogen lowers BP in female SHR. Strain differences in relaxation rates are independent of gender and estrogen status. Estrogen has no effect on arterial muscle relaxation, suggesting another mechanism for the protective effect of estrogen in hypertension.

Analysis of Variance↗

Relative changes with contraction in the central excitability state of the tibialis anterior and calf muscles.

F responses were recorded from the surface of the tibialis muscle and medial aspect of the soleus muscle in 14 normal subjects. The persistence (that is the fraction of measurable F responses found with a series of supramaximal stimuli) and average F amplitudes (measured peak-to-peak and based on at least five F responses) were determined both at rest and with isometric contraction with the ankle maintained at 90 degrees. Although the persistence at rest was significantly less in the tibialis anterior soleus than the (p less than 0.001), no significant difference was found with the muscles contracted. This was associated with a significant increase in both average F amplitudes and average F amplitude/direct motor response ratios in the tibialis anterior in comparison to the soleus. In four of the subjects, studies were also performed when the H reflex in the soleus muscle was eliminated by thigh compression. Comparable changes in both F response persistence and average F amplitude were found with and without an H reflex. These data indicate that, in contrast to the situation at rest, with isometric contraction the "central excitatory state" of the tibialis anterior is at least as great as in its antagonist antigravity muscles and that this is not due simply to increased large fiber reflex input associated with agonist contraction.

Adult↗

Muscle soreness and intramuscular fluid pressure: comparison between eccentric and concentric load.

This study investigates the dynamic and resting intramuscular pressures associated with eccentric and concentric exercise of muscles in a low-compliance compartment. The left and righ leg anterior compartments of eight healthy males (ages 22-32 yr) were exercised by either concentric or eccentric contractions of the same load (400 submaximal contractions at constant rate, 20/min for 20 min at a load corresponding to 15% of individual maximal dorsiflexion torque). Tissue fluid pressures were measured with the slit-catheter technique before, during, and after the exercise. Average peak intramuscular pressure generated during eccentric exercise (236 mmHg) was significantly greater than during concentric exercise (157 mmHg, P less than 0.001). Peak isometric contraction pressure in the eccentrically exercised compartment was significantly higher both within 20 min postexercise and on the second postexercise day (P less than 0.001). Resting pressure 2 days postexercise was significantly higher on the eccentrically exercised side (10.5 mmHg) compared with the concentrically exercised (4.4 mmHg, P less than 0.05). The ability to sustain tension during postexercise isometric contractions was impaired on the "eccentric" side. Soreness was exclusively experienced in the eccentrically exercised muscles. We conclude that eccentric exercise causes significant intramuscular pressure elevation in the anterior compartment, not seen following concentric exercise, and that this may be one of the factors associated with development of delayed muscle soreness in a tight compartment.

Adult↗

Force measurements by micromanipulation of a single actin filament by glass needles.

Single actin filaments (approximately 7 nm in diameter) labelled with fluorescent phalloidin can be clearly seen by video-fluorescence microscopy. This technique has been used to observe motions of single filaments in solution and in several in vitro movement assays. In a further development of the technique, we report here a method to catch and manipulate a single actin filament (F-actin) by glass microneedles under conditions in which external force on the filament can be applied and measured. Using this method, we directly measured the tensile strength of a filament (the force necessary to break the bond between two actin monomers) and the force required for a filament to be moved by myosin or its proteolytic fragment bound to a glass surface in the presence of ATP. The first result shows that the tensile strength of the F-actin-phalloidin complex is comparable with the average force exerted on a single thin filament in muscle fibres during isometric contraction. This force is increased only slightly by tropomyosin. The second measurement shows that the myosin head (subfragment-1) can produce the same ATP-dependent force as intact myosin. The magnitude of this force is comparable with that produced by each head of myosin in muscle during isometric contraction.

Actin Cytoskeleton↗

Force and contraction time relation in skeletal muscles of children and adults.

The force and contraction time relation was studied in twitch isometric contraction of flexor carpi ulnaris muscle (FCU) and triceps surae (TS) in 5-week-old infants, 3-year-old children and adults. Contraction time (CT), duration of the plateau of active state (PAS), and time of the decline of active state (DAS) do not change with increasing force of contraction of FCU in all three age groups and in TS in 5-week-old infants. CT and DAS decrease with increasing force of contraction in 3-year-old children and in adults. PAS remains unchanged with increasing force of TS in 3-year-old children and decreases in adults, but considerably less than CT and DAS. Evidence is adduced supporting the view that CT changes with increasing force of TS depend mainly on the elastic properties of the muscle.

Adult↗

Selective fatigue of fast motor units after electrically elicited muscle contractions.

The aim of the present study was to elucidate the electrophysiological manifestations of selective fast motor unit (MU) activation by electrical stimulation (ES) of knee extensor muscles. In six male subjects, test contraction measurement at 40% maximal voluntary contraction (MVC) was performed before and at every 5 min (5, 10, 15 and 20 min) during 20-min low intensity intermittent exercise of either ES or voluntary contractions (VC) at 10% MVC (5-s isometric contraction and 5-s rest cycles). Both isolated intramuscular MU spikes obtained from three sets of bipolar fine-wire electrodes and surface electromyogram (EMG) were simultaneously recorded and were analyzed by means of a computer-aided intramuscular spike amplitude-frequency analysis and frequency power spectral analysis, respectively. Results indicated that mean MU spike amplitude, particularly those MUs with relatively large amplitude, was significantly reduced while those MUs with small spike amplitude increased their firing rate during the 40% MVC test contraction after the ES. This was accompanied by the increased amplitude of surface EMG (rmsEMG). However, no such significant changes in the intramuscular and surface EMGs were observed after VC. These findings indicated differential MU activation patterns in terms of MU recruitment and rate coding characteristics during ES and VC, respectively. Our data strongly suggest the possibility of "an inverse size principle" of MU recruitment during ES.

Action Potentials↗