Changes In Contractile Properties Research Articles

Age- and Gender-Related Changes in Contractile Properties of Non-Atrophied EDL Muscle

BackgroundIn humans, ageing causes skeletal muscles to become atrophied, weak, and easily fatigued. In rodent studies, ageing has been associated with significant muscle atrophy and changes in the contractile properties of the muscles. However, it is not entirely clear whether these changes in contractile properties can occur before there is significant atrophy, and whether males and females are affected differently.Methods and ResultsWe investigated various contractile properties of whole isolated fast-twitch EDL muscles from adult (2–6 months-old) and aged (12–22 months-old) male and female mice. Atrophy was not present in the aged mice. Compared with adult mice, EDL muscles of aged mice had significantly lower specific force, longer tetanus relaxation times, and lower fatiguability. In the properties of absolute force and muscle relaxation times, females were affected by ageing to a greater extent than males. Additionally, EDL muscles from a separate group of male mice were subjected to eccentric contractions of 15% strain, and larger force deficits were found in aged than in adult mice.ConclusionOur findings provide further insight into the muscle atrophy, weakness and fatiguability experienced by the elderly. We have shown that even in the absence of muscle atrophy, there are definite alterations in the physiological properties of whole fast-twitch muscle from ageing mice, and for some of these properties the alterations are more pronounced in female mice than in male mice.

Changes in contractile properties of skinned single rat soleus and diaphragm fibres after chronic hypoxia

Hypoxia may be one of the factors underlying muscle dysfunction during ageing and chronic lung and heart failure. Here we tested the hypothesis that chronic hypoxia per se affects contractile properties of single fibres of the soleus and diaphragm muscle. To do this, the force-velocity relationship, rate of force redevelopment and calcium sensitivity of single skinned fibres from normoxic rats and rats exposed to 4 weeks of hypobaric hypoxia (410 mmHg) were investigated. The reduction in maximal force (P(0)) after hypoxia (p=0.031) was more pronounced in type IIa than type I fibres and was mainly attributable to a reduction in fibre cross-sectional area (p=0.044). In type IIa fibres this was aggravated by a reduction in specific tension (p=0.001). The maximal velocity of shortening (V (max)) and shape of the force velocity relation (a/P(0)), however, did not differ between normoxic and hypoxic muscle fibres and the reduction in maximal power of hypoxic fibres (p=0.012) was mainly due to a reduction in P(0). In conclusion, chronic hypoxia causes muscle fibre dysfunction which is not only due to a loss of muscle mass, but also to a diminished force generating capacity of the remaining contractile material. These effects are similar in the soleus and diaphragm muscle, but more pronounced in type IIa than I fibres.

Temporal Adaptive Changes in Contractility and Fatigability of Diaphragm Muscles from Streptozotocin-Diabetic Rats

Diabetes is characterized by ventilatory depression due to decreased diaphragm (DPH) function. This study investigated the changes in contractile properties of rat DPH muscles over a time interval encompassing from 4 days to 14 weeks after the onset of streptozotocin-induced diabetes, with and without insulin treatment for 2 weeks. Maximum tetanic force in intact DPH muscle strips and recovery from fatiguing stimulation were measured. An early (4-day) depression in contractile function in diabetic DPH was followed by gradual improvement in muscle function and fatigue recovery (8 weeks). DPH contractile function deteriorated again at 14 weeks, a process that was completely reversed by insulin treatment. Maximal contractile force and calcium sensitivity assessed in Triton-skinned DPH fibers showed a similar bimodal pattern and the same beneficial effect of insulin treatment. While an extensive analysis of the isoforms of the contractile and regulatory proteins was not conducted, Western blot analysis of tropomyosin suggests that the changes in diabetic DPH response depended, at least in part, on a switch in fiber type.

Possible Mechanisms for the Effect of Protein Sensitization on Contractile Function of Fast and Slow Muscles in Mice

We studied the mechanisms underlying the effect of immunobiological reorganization of the organism on contractile function of isolated skeletal muscles from mouse leg (fast muscle, m. extensor digitorum longus; and slow muscle, m. soleus). Protein sensitization was accompanied by changes in contractile properties of fast and slow skeletal muscles. These changes were differently directed in muscles with various phenotypes. The force of carbachol-induced contraction (cholinergic agonist) increased in the slow muscle, but decreased in the fast muscle. The direction of changes in the force of carbachol-induced contractions under conditions of protein sensitization in skeletal muscles correlates with changes in non-quantal secretion of acetylcholine in the endplate (H-effect). Opposite changes in functional properties of fast and slow muscles from mouse leg during protein sensitization are related to choline-mediated excitation of the muscle fiber membrane. Our results suggest that changes in contractile function of skeletal muscles during protein sensitization are associated with variations in choline-mediated excitation of the muscle fiber membrane and modification of electromechanical coupling.

Altered contractile properties of the gastrocnemius muscle poststroke

Spasticity, contracture and muscle weakness often occur together poststroke and cause considerable motor impairments to stroke survivors. The underlying changes in contractile properties of muscle fascicles are still not clear. The purpose of this study was to investigate the contractile property changes of the medial gastrocnemius muscle fascicles poststroke. Ten stroke survivors and ten healthy subjects participated in the study. The medial gastrocnemius fascicular length was measured at various combinations of ankle and knee positions using ultrasonography, with the muscle activated selectively using electrical stimulation. The stimulation intensity was kept constant across different ankle and knee positions to establish the active force-length relationship of the muscle fascicles. It was found that stroke survivors showed a shift of the force-length curve with a significantly shorter optimal fascicle length (33.2 +/- 3.2 mm) compared with that of healthy controls (47.4 +/- 2.7 mm) with P < 0.001. Furthermore, the width span of the fascicular force-length curve of stroke survivors was significantly narrower with steeper slopes than that of controls (P <or= 0.001), suggesting reduced number of sarcomeres along the fascicles and/or reduced sarcomere length poststroke. Regression analysis showed that the medial gastrocnemius fascicular length of stroke survivors varied significantly less with ankle and knee flexions (P <or= 0.001) than that of controls, suggesting shorter and stiffer muscle fascicles poststroke, which might be attributed to muscle architectural adaptation. This study showed that there are considerable changes in the contractile properties of muscle fascicles poststroke, which may contribute directly to the joint-level changes of decreased range of motion, increased stiffness, muscle weakness, and impaired motor functions in stroke survivors.

Effect of thyroid hormone on the contractility of self-organized heart muscle

Tissue-engineered heart muscle may provide an alternative treatment modality for end-stage congestive heart failure. We have previously described a method to engineer contractile heart muscle in vitro (termed cardioids). This study describes a method to improve the contractile properties of cardioids utilizing thyroid hormone (T3) stimulation. Cardioids were engineered by promoting the self-organization of primary neonatal cardiac cells into a contractile tissue construct. Cardioids were maintained in standard cell culture media supplemented with varying concentrations of T3 in the range 1-5ng/ml. The contractile properties of the cardioids were evaluated 48h after formation. Stimulation with T3 resulted in an increase in the specific force of cardioids from an average value of 0.52 +/- 0.16kPa (N = 6) for control cardioids to 2.42 +/- 0.29kPa (N = 6) for cardioids stimulated with 3ng/ml T3. In addition, there was also an increase in the rate of contraction and relaxation in response to T3 stimulation. Cardioids that were stimulation with T3 exhibited improved pacing characteristics in response to electrical pacing at 1-5Hz and an increase in the degree of spontaneous contractility. Changes in the gene expression of SERCA2, phospholamban, alpha-myosin heavy chain, and beta-myosin heavy chain correlated with the changes in contractile properties. This study demonstrates the modulation of the contractile properties of tissue-engineered heart muscle using T3 stimulation.

Adaptive responses to creatine loading and exercise in fast-twitch rat skeletal muscle

We investigated the effects of chronic creatine loading and voluntary running (Run) on muscle fiber types, proteins that regulate intracellular Ca2+, and the metabolic profile in rat plantaris muscle to ascertain the bases for our previous observations that creatine loading results in a higher proportion of myosin heavy chain (MHC) IIb, without corresponding changes in contractile properties. Forty Sprague-Dawley rats were assigned to one of four groups: creatine-fed sedentary, creatine-fed run-trained, control-fed sedentary, and control-fed run-trained animals. Proportion and cross-sectional area increased 10% and 15% in type IIb fibers and the proportion of type IIa fibers decreased 11% in the creatine-fed run-trained compared with the control-fed run-trained group (P < 0.03). No differences were observed in fast Ca2+-ATPase isoform SERCA1 content (P > 0.49). Creatine feeding alone induced a 41% increase (P < 0.03) in slow Ca2+-ATPase (SERCA2) content, which was further elevated by 33% with running (P < 0.02). Run training alone reduced parvalbumin content by 50% (P < 0.05). By comparison, parvalbumin content was dramatically decreased by 75% (P < 0.01) by creatine feeding alone but was not further reduced by run training. These adaptive changes indicate that elevating the capacity for high-energy phosphate shuttling, through creatine loading, alleviates the need for intracellular Ca2+ buffering by parvalbumin and increases the efficiency of Ca2+ uptake by SERCAs. Citrate synthase and 3-hydroxyacyl-CoA dehydrogenase activities were elevated by run training (P < 0.003) but not by run training + creatine feeding. This indicates that creatine loading during run training supports a faster muscle phenotype that is adequately supported by the existing glycolytic potential, without changes in the capacity for terminal substrate oxidation.

Effects of sex hormones on genioglossal muscle contractility and SR Ca 2+-ATPase activity in aged rat

This investigation was designed to examine whether short-term administration of sex hormones could produce changes in contractile properties and the sarcoplasmic reticulum Ca 2+-ATPase (SERCA) function in genioglossal muscle (GG) of aged male rats. Twenty-four aged male rats were randomly divided into three groups to receive an intramuscular injection of either 0.1 mg/kg oestrogen (group A), 2.5 mg/kg testosterone (group B), or 0.2 ml sterile peanut oil (group C, control), twice a week, during 4 weeks. After hormone treatment, in vivo isometric contractile properties were determined using surgically prepared GG muscles with platinum electrodes for stimulation of the medial branch of the hypoglossus nerve. Sarcoplasmic reticulum Ca 2+-ATPase activity was measured in muscle homogenates by detecting the amount of inorganic phosphorus ion released in a standard coupled enzyme assay. SERCA1 mRNA level was observed using a real-time quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR). It was found that, compared with group C, testosterone treatment decreased the fatigue resistance in GG muscles ( p < .05), whereas no change was observed in the isometric twitch ( P t) and tetanic tension ( P 0) ( p > .05). By contrast, in oestrogen treated GG muscles, no significant modification was found either in the contractile or in endurance properties ( p > .05). The change in GG fatigue resistance of group B was associated with a marked decrease in SR Ca 2+-ATPase activity when compared with that of the control group ( p < .01). Furthermore, SERCA1 mRNA level was also down regulated in group B ( p < .05). No prominent differences in SR Ca 2+-ATPase activity and SERCA1 mRNA expression existed between group A and group C ( p > .05). The present results show that exogenous testosterone produces more pronounced changes in GG muscle fatigue resistance than oestrogen does by acting at SR Ca 2+-ATPase activity and SERCA gene expression.

Age-related resistance to skeletal muscle fatigue is preserved during ischemia

During voluntary contractions, the skeletal muscle of healthy older adults often fatigues less than that of young adults, a result that has been explained by relatively greater reliance on muscle oxidative metabolism in the elderly. Our aim was to investigate whether this age-related fatigue resistance was eliminated when oxidative metabolism was minimized via ischemia induced by cuff (220 mmHg). We hypothesized that 1) older men (n = 12) would fatigue less than young men (n = 12) during free-flow (FF) contractions; 2) both groups would fatigue similarly during ischemia; and 3) reperfusion would reestablish the fatigue resistance of the old. Subjects performed 6 min of intermittent, maximal voluntary isometric contractions of the ankle dorsiflexors under FF and ischemia-reperfusion (IR) conditions. Ischemia was maintained for the first 3 min of contractions, followed by rapid cuff deflation and reperfusion for 3 additional minutes of contractions. Central activation, peripheral activation, and muscle contractile properties were measured at 3 and 6 min of contractions. Older men fatigued less than young men during FF (P </= 0.02), ischemia (P < 0.001), and reperfusion (P < 0.001). During FF, activation and contractile properties changed similarly across age groups. At the end of ischemia, central (P = 0.02) and peripheral (P </= 0.03) activation declined more in the young, with no effect of age on the changes in contractile properties. Thus age-related fatigue resistance was evident during FF and IR, indicating that differences in blood flow and oxidative metabolism do not explain the fatigue resistance of old age.

Change in contractile properties of human muscle in relationship to the loss of power and slowing of relaxation seen with fatigue.

Slow relaxation from an isometric contraction is characteristic of acutely fatigued muscle and is associated with a decrease in the maximum velocity of unloaded shortening (V(max)) and both these phenomena might be due to a decreased rate of cross bridge detachment. We have compared the change in relaxation rate with that of various parameters of the force-velocity relationship over the course of an ischaemic series of fatiguing contractions and subsequent recovery using the human adductor pollicis muscle working in vivo at approximately 37 degrees C in nine healthy young subjects. Maximal isometric force (F(0)) decreased from 91.0 +/- 1.9 to 58.3 +/- 3.5 N (mean +/- s.e.m.). Maximum power decreased from 53.6 +/- 4.0 to 17.7 +/- 1.2 (arbitrary units) while relaxation rate declined from -10.3 +/- 0.38 to -2.56 +/- 0.29 s(-1). V(max) showed a smaller relative change from 673 +/- 20 to 560 +/- 46 deg s(-1) and with a time course that differed markedly from that of slowing of relaxation, showing very little change until late in the series of contractions. Curvature of the force-velocity relationship increased (a/F(0) decreasing from 0.22 +/- 0.02 to 0.11 +/- 0.02) with fatigue and with a time course that was similar to that of the loss of power and the slowing of relaxation. It is concluded that for human muscle working at a normal physiological temperature the change in curvature of the force-velocity relationship with fatigue is a major cause of loss of power and may share a common underlying mechanism with the slowing of relaxation from an isometric contraction.

Changes in contractile properties of motor units of the rat medial gastrocnemius muscle after spinal cord transection

The effects of complete transection of the spinal cord at the level of Th9/10 on contractile properties of the motor units (MUs) in the rat medial gastrocnemius (MG) muscle were investigated. Our results indicate that 1 month after injury the contraction time (time-to-peak) and half-relaxation time were prolonged and the maximal tetanic force in most of the MUs in the MG muscle of spinal rats was reduced. The resistance to fatigue also decreased in most of the MUs in the MG of spinal animals. Moreover, the post-tetanic potentiation of twitches in MUs diminished after spinal cord transection. Criteria for the division of MUs into three types, namely slow (S), fast fatigue resistant (FR) and fast fatigable (FF), applied in intact animals, could not be directly used in spinal animals owing to changes in contractile properties of MUs. The 'sag' phenomenon observed in unfused tetani of fast units in intact animals essentially disappeared in spinal rats and it was only detected in few units, at low frequencies of stimulation only. Therefore, the MUs in spinal rats were classified as fast or slow on the basis of an adjusted borderline of 20 ms, instead of 18 ms as in intact animals, owing to a slightly longer contraction time of those fast motor units with the 'sag'. We conclude that all basic contractile properties of rat motor units in the medial gastrocnemius muscle are significantly changed 1 month after complete spinal cord transection, with the majority of motor units being more fatigable and slower than those of intact rats.

Contractile properties and myosin heavy chain composition of rat tongue retrusor musculature show changes in early adulthood after 19 days of artificial rearing

Previously, we showed that artificial rearing using the "pup in a cup" model results in decreased tongue activity and caused some minor alterations in the tongue retrusor musculature. However, the artificial rearing time frame previously chosen was brief (11 days). The purpose of the present investigation was to extend the artificial rearing period from postnatal days 3 to 21 (P21) to determine whether significant alterations occur as a result of this reduced tongue use. Several changes in contractile properties due to the artificial rearing process were observed, which fully recovered by postnatal days 41 to 42 (P41-2). These changes included a shorter twitch contraction time, shorter twitch half-relaxation time, and decreased fatigue resistance. Styloglossus muscle exhibited more neonatal myosin heavy chain (MHC) isoform at P21 for the artificially reared (AR) group. Changes that were persistent at P41-2 were also observed. Maximum tetanic tension was lower for the AR group at P21 and P41-2 compared with their dam-reared counterparts. Twitch tension was also lower by P41-2 in the AR group. At P41-2, the AR group exhibited an increase in MHC IIa and a decrease in MHC IIb for the styloglossus muscle. In addition, the AR group exhibited a decreased MHC IIb for the long head of the biceps brachii at P41-2. Our results are similar to other models of hindlimb immobilization and suspension. By extending our artificial rearing period, this reduced tongue activity induced acute changes and alterations in the tongue retrusor musculature that persisted into early adulthood.

Muscle Fatigue during High-Intensity Exercise in Children

Children are able to resist fatigue better than adults during one or several repeated high-intensity exercise bouts. This finding has been reported by measuring mechanical force or power output profiles during sustained isometric maximal contractions or repeated bouts of high-intensity dynamic exercises. The ability of children to better maintain performance during repeated high-intensity exercise bouts could be related to their lower level of fatigue during exercise and/or faster recovery following exercise. This may be explained by muscle characteristics of children, which are quantitatively and qualitatively different to those of adults. Children have less muscle mass than adults and hence, generate lower absolute power during high-intensity exercise. Some researchers also showed that children were equipped better for oxidative than glycolytic pathways during exercise, which would lead to a lower accumulation of muscle by-products. Furthermore, some reports indicated that the lower ability of children to activate their type II muscle fibres would also explain their greater resistance to fatigue during sustained maximal contractions. The lower accumulation of muscle by-products observed in children may be suggestive of a reduced metabolic signal, which induces lower ratings of perceived exertion. Factors such as faster phosphocreatine resynthesis, greater oxidative capacity, better acid-base regulation, faster readjustment of initial cardiorespiratory parameters and higher removal of metabolic by-products in children could also explain their faster recovery following high-intensity exercise.From a clinical point of view, muscle fatigue profiles are different between healthy children and children with muscle and metabolic diseases. Studies of dystrophic muscles in children indicated contradictory findings of changes in contractile properties and the muscle fatigability. Some have found that the muscle of boys with Duchenne muscular dystrophy (DMD) fatigued less than that of healthy boys, but others have reported that the fatigue in DMD and in normal muscle was the same. Children with glycogenosis type V and VII and dermatomyositis, and obese children tolerate exercise weakly and show an early fatigue. Studies that have investigated the fatigability in children with cerebral palsy have indicated that the femoris quadriceps was less fatigable than that of a control group but the fatigability of the triceps surae was the same between the two groups. Further studies are required to elucidate the mechanisms explaining the origins of muscle fatigue in healthy and diseased children. The use of non-invasive measurement tools such as magnetic resonance imaging and magnetic resonance spectroscopy in paediatric exercise science will give researchers more insight in the future.

Effect of joint angle on mechanomyographic amplitude during unfused and fused tetani in the human biceps brachii muscle

The influence of muscle length-dependent changes in contractile properties on the mechanomyogram (MMG) was investigated during evoked contractions of the biceps brachii muscle. The biceps of nine healthy subjects was stimulated by single twitches, unfused (10 Hz), and fused (30 Hz) frequencies at elbow joint angles of 75, 90, 105, 120, 135, and 150 degrees. During evoked contractions, the longitudinal movement was estimated by the fluctuation of torque signal, and the lateral movement was detected by MMG. Contraction time and half relaxation time were calculated from the single twitches. For repetitive stimulations, followed by eliminating DC component from torque signals, the root mean square values calculated from the torque and MMG signals, which were torque fluctuation and RMS-MMG, respectively. For the 10 Hz stimulation condition, reductions in the torque fluctuation and RMS-MMG were concomitantly observed with increasing elbow angle, and there was a significant correlation between the torque fluctuation and RMS-MMG. On the other hand, for 30 Hz stimulation, there were no significant differences in the torque fluctuation and RMS-MMG over all elbow angles, and no significant correlation between the two parameters. Moreover, the torque fluctuation and RMS-MMG for 10 Hz stimulation were correlated with the contraction time and half relaxation time obtained at each elbow angle, while there were no correlations for the 30 Hz condition. These data suggest that MMG could be a reliable tool to study the development of fusion and the changes in muscle contractile properties during repetitive unfused contractions.

Beta2-agonist administration increases sarcoplasmic reticulum Ca2+-ATPase activity in aged rat skeletal muscle.

Aging is associated with a slowing of skeletal muscle contractile properties, including a decreased rate of relaxation. In rats, the age-related decrease in the maximal rate of relaxation is reversed after 4-wk administration with the beta2-adrenoceptor agonist (beta2-agonist) fenoterol. Given the critical role of the sarcoplasmic reticulum (SR) in regulating intracellular Ca2+ transients and ultimately the time course of muscle contraction and relaxation, we tested the hypothesis that the mechanisms of action of fenoterol are mediated by alterations in SR proteins. Sarcoendoplasmic reticulum Ca2+-ATPase (SERCA) kinetic properties were assessed in muscle homogenates and enriched SR membranes isolated from the red (RG) and white (WG) portions of the gastrocnemius muscle in adult (16 mo) and aged (28 mo) F344 rats that had been administered fenoterol for 4 wk (1.4 mg/kg/day ip, in saline) or vehicle only. Aging was associated with a 29% decrease in the maximal activity (Vmax) of SERCA in the RG but not in the WG muscles. Fenoterol treatment increased the Vmax of SERCA and SERCA1 protein levels in RG and WG. In the RG, fenoterol administration reversed an age-related selective nitration of the SERCA2a isoform. Our findings demonstrate that the mechanisms underlying age-related changes in contractile properties are fiber type dependent, whereas the effects of fenoterol administration are independent of age and fiber type.

Changes in contractile properties and neuromuscular propagation evaluated by simultaneous mechanomyogram and electromyogram during experimentally induced hypothermia

The present study examined, whether or not mechanomyogram (MMG) amplitude and frequency component could reflect the contractile properties of the triceps surae muscles, composed of relatively slow soleus (SOL) and fast medial gastrocnemius (MG), during experimentally induced hypothermia condition. In eight male subjects, lying in prone position, supramaximal single twitch and repetitive electrical stimulations at 10 Hz were applied at the intramuscular temperatures of control (34 °C), 15, 20, and 25 °C, respectively. The hypothermia induced substantial reduction in muscle contractile properties, e.g. prolonged twitch contraction and half relaxation times, resulted in a highly significant reduction in the fluctuation of force signal during the repetitive stimulations. These changes were almost mirrored by the similar and significant reductions in the MMG amplitude in both SOL and MG. Power spectrum analysis revealed that peak frequency components of MMG and fluctuation of force were almost matched with the applied stimulation frequencies, independent of the temperature condition. These results strongly suggest that MMG analysis could be employed to study muscle contractile properties varying across different physiological conditions.

Increased antioxidant capacity does not attenuate muscle atrophy caused by unweighting.

Previous studies have increased antioxidant capacity in skeletal muscle to attenuate oxidative stress and muscle atrophy during limb immobilization (Appell HJ, Duarte JAR, and Soares JMC. Int J Sports Med 18: 157-160, 1997; Kondo H, Miura M, Nakagaki I, Sasaki S, and Itokawa Y. Am J Physiol Endocrinol Metab 262: E583-E590, 1992). The purpose of this study was to determine the level of oxidative stress in muscle during hindlimb unweighting (HLU) and whether antioxidant supplementation can attenuate the atrophy and changes in contractile properties resulting from 14 days of unweighting. Muscle unweighting caused a 44% decrease in soleus (Sol) and a 30% decrease in gastrocnemius (GS) mass, a 7% decrease in body weight, and 28% decrease in tetanic force in the GS. Protein carbonyls increased by 44% in the Sol with HLU. Antioxidant supplementation did not attenuate the GS or Sol atrophy or the decrease in GS force generation during HLU. Sol and GS protein concentration was not different between groups. The GS was also subjected to three different oxidative challenges to determine whether the supplement increased the antioxidant capacity of the muscle. In all cases, muscles exhibited an increased antioxidant capacity. These data indicate that antioxidant supplementation was not an effective countermeasure to the atrophy associated with HLU.

Changes in contractile properties of skeletal muscle during developmentally programmed atrophy and death.

Skeletal muscle atrophy and death are protracted processes that accompany aging and pathological insults in mammals. The intersegmental muscles (ISMs) from the tobacco hawkmoth Manduca sexta are composed of giant fibers that undergo distinct hormonally-regulated programs of atrophy and death at the end of metamorphosis. Atrophy occurs during the 3 days preceding adult emergence and results in a 40% reduction of mass, whereas death takes place during the subsequent 30 h and results in the complete loss of the fibers. There are no significant changes in tetanic force or calcium sensitivity in skinned fiber preparations during atrophy. However, the size of caffeine-induced contractions fell by about 50%. With the onset of the death phase, dramatic reductions occur in ISM: tetanic force, twitch amplitude, resting potential, caffeine-induced contractions, calcium sensitivity, and Hill coefficients. Several lines of evidence suggest that ISM atrophy is caused by an increase in protein turnover without significant modification of fiber organization. In contrast, ISM death is accompanied by disorganization of the contractile apparatus and concomitant loss of contractile function.

Junctional sarcoplasmic reticulum transmembrane proteins in the heart.

The uptake of calcium into the sarcoplasmic reticulum (SR) and its subsequent release from the SR play a key role in the regulation of the cytosolic calcium concentration and the excitation-contraction coupling in cardiac muscle. While calcium uptake, catalyzed by the calcium-dependent ATPase, is thought to occur throughout the SR, the release of calcium is controlled by a complex of proteins localized to a distinct region, the junctional SR. This complex consists of the calcium release channel or ryanodine receptor (RyR), the high capacity calcium-binding protein calsequestrin located in the lumen of the junctional SR, and the junctional SR transmembrane proteins triadin 1 and junctin which are hypothesized to anchor calsequestrin to the RyR. Transgenic mice with cardiac-specific overexpression of triadin 1 or junctin show distinct cardiac phenotypes with altered cellular and subcellular morphology, changes in contractile properties and/or in the expression of junctional SR proteins suggesting that these junctional sarcoplasmic reticulum transmembrane proteins are of functional relevance for the regulation of calcium release in the heart.

Circumferential stretching of saphenous vein smooth muscle enhances vasoconstrictor responses by Rho kinase-dependent pathways.

Surgical preparation and/or pulsatile arterial perfusion of saphenous vein increases the sensitivity of vein rings to calcium mobilising agonists such as phenylephrine. We have investigated the mechanism(s) underlying this effect. We have used an ex vivo flow circuit, with simulated arterial or venous flows (mean pressure 100 and 20 mmHg, respectively), to investigate the sensitivity of human saphenous vein to phenylephrine, 5-hydroxytryptamine (5-HT) and KCl, using organ chamber pharmacology. After 90 min of pulsatile arterial perfusion the mean maximum tension induced by KCl had increased from 4.7 to 11.1 g (n=5), by phenylephrine had increased from 4.4 to 10.2 g (n=8) and by 5-HT had increased from 4.4 to 6.7 g (n=10), all P<0.01. Phenylephrine did not augment the tension in vein rings maximally precontracted with KCl (n=4). The EC(50) for KCl was unchanged after pulsatile arterial perfusion (n=5), but for phenylephrine and 5-HT there were significant reductions from 14+/-5 to 2+/-1 microM (n=8) and from 1.0+/-0.4 to 0.20+/-0.06 microM (n=10), respectively. The rate of contraction (in response to 3 microM phenylephrine) increased from 0.11 g/min to 0.37 g/min, P<0.02, after arterial perfusion (n=4). These changes in contractile properties (to phenylephrine) were endothelium-independent, evident within 5 min of simulated arterial perfusion. The changes in contractile properties could be abrogated by external stenting of the vein (to attenuate circumferential deformation) or inclusion in the perfusate of a vasodilator, e.g., cromakalim (5 microM) or the selective Rho kinase inhibitor Y-27632 (20 microM). The heightened sensitivity and contractility to phenylephrine was maintained after inclusion of adenosine (100 microM), gadolinium (10 microM) or cycloheximide (10 microM) in the vein perfusate. The circumferential deformations imposed by simulated arterial perfusion alter the vasomotor responses of saphenous vein smooth muscle. These effects are independent of new protein synthesis or the activation of stretch activated cation channels. The Rho kinase pathway appears to mediate the signalling mechanisms leading to increased agonist-induced tension and the increased sensitivity to vasoconstrictors.