Nandrolone decanoate pre-treatment attenuates unweighting-induced functional changes in rat soleus muscle.

The effect of nandrolone decanoate pre-treatment (15 mg kg(-1) week(-1), for 6 weeks) was tested on the changes in mass and contractile properties of soleus muscle associated with 3 weeks of hindlimb suspension. Male rats were assigned to four groups (eight animals/group): control, nandrolone decanoate hindlimb-loaded, hindlimb-unweighted and nandrolone decanoate hindlimb-unweighted. Compared with age-matched control values, suspension induced a reduction in relative muscle mass and a shift in tension characteristics from slow-towards fast-twitch type. Nandrolone decanoate pre-treatment of suspended animals (nandrolone decanoate hindlimb-unweighted vs. nandrolone decanoate hindlimb-loaded) partially spared the relative soleus mass. Furthermore, (1) the relative twitch tension (nandrolone decanoate hindlimb-loaded: 5.4 +/- 0.7%; nandrolone decanoate hindlimb-unweighted: 5.1 +/- 0.5%), (2) the time to peak tension (nandrolone decanoate hindlimb-loaded: 152 +/- 9 ms; nandrolone decanoate hindlimb-unweighted: 167 +/- 15 ms), (3) the time constant of relaxation (nandrolone decanoate hindlimb-loaded: 274 +/- 12 ms; nandrolone decanoate hindlimb-unweighted: 245 +/- 20 ms), (4) the relative K+ contracture tension (nandrolone decanoate hindlimb-loaded: 81.7 +/- 3.8%; nandrolone decanoate hindlimb-unweighted: 86.9 +/- 4.2%) and (5) the relative caffeine contracture tension (0.5 mM) (nandrolone decanoate hindlimb-loaded: 5.2 +/- 0.8%; nandrolone decanoate hindlimb-unweighted: 5.9 +/- 1.1%) were not significantly modified. The present results demonstrate that exogenously provided nandrolone decanoate pre-treatment attenuates functional changes occurring in soleus muscle subject to unweighting.

Nandrolone decanoate treatment induces changes in contractile responses of rat untrained fast-twitch skeletal muscle.

This investigation was designed to examine whether short-term administration of anabolic-androgenic steroids (AAS) (nandrolone decanoate) could produce changes in contractile responses of untrained rat fast- (edl) and slow- (soleus) twitch skeletal muscle. Twenty male rats were divided into two groups, one group received weekly (for 6 weeks) an intramuscular injection of AAS, nandrolone decanoate (15 mg kg(-1)) and the second group received weekly the similar doses of vehicle (sterile peanut oil). In edl intact isolated small bundles (two to four cells), it was found that nandrolone decanoate treatment increases the K+ contracture tension (146 mM) relative to maximum tension by 56%, whereas no change was observed in the time to peak tension and in the time constant of relaxation. By contrast, in treated soleus muscle, compared with control, no significant modification was found in the K+ contracture characteristics. The change in edl contractile responses was associated with a shift to more negative potential of the voltage-dependence activation and the steady-state inactivation curves which also shifted leftward in treated soleus fibres. Furthermore, in edl skinned Triton X-100 fibres, the Ca2+ sensitivity of contractile proteins (pCa50) was increased, while electrophoresis analysis indicates no significant effect of nandrolone decanoate treatment on myosin heavy chain (MHC) isoforms. The present results show that nandrolone decanoate treatment produces more pronounced changes in untrained fast muscle function rather than soleus by acting at different levels of the excitation-contraction coupling mechanism without changes in the MHC isoforms and that contractile responses became similar to those found in soleus muscle.

A comparative analysis of the effects of exercise training on contractile responses in fast- and slow-twitch rat skeletal muscles.

The effects of 5 weeks treadmill-exercise training on isometric tension and contractile proteins were studied in intact and skinned isolated small bundles of rat skeletal soleus and extensor digitorum longus (edl) fibers. In soleus and edl muscles, 5 weeks exercise training: (i) increased twitch amplitude by 25% and 8%, respectively, without modification in the time-to-peak tension and the time constant of relaxation, (ii) increased the amplitude of K(+) contracture by 93% and 88%, respectively, and accelerated its relaxation by 17% and 43%, respectively, and (iii) increased the amplitude of caffeine contractures (soleus: 0.5 mM: 86%, 10 mM: 77%; edl: 0.5 mM: 89%, 10 mM: 87%). In conclusion, changes in contractile responses were associated with shifts in the steady state inactivation curves and in the voltage-dependent activation curve to a more negative potential, with increases in soleus and edl caffeine sensitivity, without changes in the Ca(2+) sensitivity of contractile proteins and myosin heavy chain isoforms.

Inhibition of caffeine-induced Ca2+ release by adenosine in mammalian skinned slow- and fast-twitch fibres.

The present study performed on chemically skinned skeletal fibres was designed to compare the effects of adenosine on the Ca2+ sensitivity of contractile proteins and on caffeine-induced Ca2+ release in rat slow- (soleus) and fast-twitch (edl) muscles. The tension-pCa relationships were obtained by exposing triton X-100 (1% v/v) skinned fibres sequentially to solutions of decreasing pCa in the presence or in absence of adenosine. Then, changes in caffeine contracture due to adenosine were recorded on saponin (50 microg/ml) skinned fibres. The results show that the sensitivity to Ca2+ of contractile proteins in the presence of different concentrations of caffeine was not significantly modified by adenosine. However, it was proposed that adenosine (0.1-2 mM) reduced the Ca2+ released by caffeine (0.1-10 mM) from the sarcoplasmic reticulum in slow- and fast-twitch fibres and that the soleus was more sensitive to adenosine than edl muscle. The effects of specific A2a and A1 agonists and antagonists were also tested on caffeine contractures. It was found that the A1 antagonist reduced adenosine effect on caffeine response. Then it is proposed that adenosine modulates the sarcoplasmic reticulum Ca2+ release by a direct effect on the RyR1 receptors and/or by an indirect effect mediated by A1 receptors located at the sarcoplasmic level.

Caffeine stimulates the reverse mode of Na/Ca2+ exchanger in ferret ventricular muscle.

This study investigated the effect of caffeine on the sarcolemmal mechanisms involved in intracellular calcium control. Ferret cardiac preparations were treated with ryanodine and thapsigargin in order to eliminate the sarcoplasmic reticulum (SR) function. This treatment abolished caffeine contracture irreversibly in normal solution. The perfusion with K-free medium that blocked the Na+--K+ pump resulted in a recovery of slow relaxing caffeine contractures similar to Na-free contractures. The amplitude of caffeine contractures was dependent on the bathing [caffeine]o and [Ca2+]o. Divalent cations Ni2+ and Cd2+, which have an inhibitory effect on the Na+/Ca2+ exchanger, produced dose-dependent inhibition of caffeine responses with apparent Ki of 780 +/- 19 and 132 +/- 5 microM, respectively. Caffeine also caused dose-dependent inhibition of Na-free contractures (Ki=4.62 +/- 1.5 mM), and the reduction or removal of [Na+]o exerted an inhibitory effect on caffeine contractures (Ki=73.5 +/- 17.12 mM). These experiments indicate that the increase in resting tension following exposure to caffeine was mediated by Na+/Ca2+ exchanger, which represents an additional element of complexity in caffeine action on cardiac muscle.

Skeletal muscle disuse induces fibre type-dependent enhancement of Na(+) channel expression.

Slow-twitch and fast-twitch muscle fibres have specific contractile properties to respond to specific needs. Since sodium current density is higher in fast-twitch than in slow-twitch fibres, sodium channels contribute to the phenotypic feature of myofibres. Phenotype determination is not irreversible: after periods of rat hindlimb unloading (HU), a model of hypogravity, a slow-to-fast transition occurs together with atrophy in the antigravity slow-twitch soleus muscle. Using cell-attached patch-clamp and northern blot analyses, we looked at sodium channel expression in soleus muscles after 1-3 weeks of HU in rats. We found that sodium channels in fast-twitch flexor digitorum brevis muscle fibres, soleus muscle fibres and 1- to 3-week HU soleus muscle fibres showed no difference in unitary conductance, open probability and voltage-dependencies of activation, fast inactivation and slow inactivation. However, muscle disuse increased sodium current density in soleus muscle fibres 2-fold, 2.5-fold and 3-fold after 1, 2 and 3 weeks of HU, respectively. The concentration of mRNA for the skeletal muscle sodium channel alpha subunit increased 2-fold after 1 week of HU but returned to the control level after 3 weeks of HU. In contrast, the concentration of mRNA for the ubiquitous sodium channel beta(1) subunit was unchanged after 1 week and had increased by 30% after 3 weeks of HU. The tetrodotoxin sensitivity of sodium currents in 3-week HU soleus muscles and the lack of mRNA signal for the juvenile skeletal muscle sodium channel alpha subunit excluded denervation in our experiments. The observed increase in sodium current density may reduce the resistance to fatigue of antigravity muscle fibres, an effect that may contribute to muscle impairment in humans after space flight or after long immobilization.

Differential effects of nandrolone decanoate in fast and slow rat skeletal muscles.

PURPOSE: We studied the effects of high doses of an anabolic-androgenic steroid, exercise training, and a combination of steroid and training on mammalian fast- and slow-twitch skeletal muscles at the cellular level. METHODS: Thirty-two male rats were divided into sedentary and treadmill-trained groups (increased speed and time: 18 m.min-1, 0.5 h.d-1, 5 d.wk-1). Eight animals of each group were treated with nandrolone decanoate (ND) (15 mg.kg-1.wk-1), and others received the same doses of solvent. The animals were killed after 5 wk, and the contractile parameters for isolated small bundles of soleus and extensor digitorum longus (edl) fibers were estimated. RESULTS: Muscle mass, twitches, and K+ contractures were increased in soleus and edl muscles after the drug treatment and after the exercise training. Caffeine contractures were increased only after the exercise training. The combination of exercise with ND treatment produced greater effects, particularly a significant increase in sensitivity to caffeine and the amplitude of K+ contractures as well as a shortening of the time required to restore contracture. These modifications were more marked in slow than fast muscle. CONCLUSION: These results show that 5 wk of exercise training produced changes in the contractile responses developed by isolated skeletal muscle cells. The combination of exercise training with ND treatment potentiated these effects, suggesting that there was some modification in the excitation-contraction coupling mechanism. ND treatment also produced a more potent effect in soleus than edl sedentary muscle.

Cyclopiazonic acid and thapsigargin reduce Ca2+ influx in frog skeletal muscle fibres as a result of Ca2+ store depletion.

We have investigated the influence of the sarcoplasmic reticulum (SR) Ca2+ content on the retrograde control of skeletal muscle L-type Ca2+ channels activity by ryanodine receptors (RyR). The effects of cyclopiazonic acid (CPA) and thapsigargin (TG), two structurally unrelated inhibitors of SR Ca(2+)-adenosine triphosphatase (ATPase), were examined on the SR Ca2+ content, the calcium current and contraction in single frog semitendinosus fibres using the double mannitol-gap technique. At moderate concentrations that only partially inhibited Ca2+ sequestration by the SR, CPA (2-4 microM) induces a concentration dependent depression of contraction and Ca2+ current amplitudes. When Ba2+ is the charge carrier, the inward current is not changed by CPA suggesting that this Ca(2+)-pump inhibitor does not directly affect dihydropyridine Ca2+ channels. Similar effects were obtained with TG (1-5 microM). Changes in Ca2+ currents and contraction were accompanied by a reduced Ca2+ loading of the SR. We attribute the modulation of the Ca2+ current to the selective inhibition of the SR Ca2+ ATPase, resulting in a decreased Ca2+ release and thereby a reduced activation of calcium inward currents. This is therefore taken to represent a calcium release-dependent modulation of skeletal muscle L-type Ca2+ channels.

Sarcoplasmic reticulum Ca(2+) content affects 4-CmC and caffeine contractures of rat skinned skeletal muscle fibers.

This study investigated whether the sarcoplasmic reticulum Ca(2+) content of rat skeletal muscle fibers affected contractile responses obtained by an application of 4-chloro-m-cresol (4-CmC) and caffeine. Contractures were elicited on saponin-skinned fibers under different Ca(2+) loading conditions. The amplitude of 4-CmC and caffeine contractures of fast-twitch muscle fibers (edl, extensor digitorum longus) differed between the different loading conditions, and this is associated with a greater change in sensitivity to 4-CmC. When the sarcoplasmic reticulum was loaded with a low Ca(2+) concentration for a short period, the 4-CmC concentration providing half-maximal response was tenfold higher than with a larger sarcoplasmic reticulum Ca(2+) loading for a longer period, whereas for caffeine this concentration was only twofold higher in the same conditions. These findings indicate that 4-CmC contractile responses of edl muscle fibers are more dependent on luminal Ca(2+) activity than those of caffeine are. Thus 4-CmC would appear to be of greater interest than caffeine for the study of muscle contractile responses where variations in intracellular Ca(2+) activity exist.

Differential effects of 4-chloro-m-cresol and caffeine on skinned fibers from rat fast and slow skeletal muscles.

Contractile responses to 4-chloro-m-cresol (4-CmC) were tested in saponin- and Triton X-100-skinned fibers from soleus and edl (extensor digitorum longus) muscles of adult rats and compared with those to caffeine. The testing of different concentrations of 4-CmC on saponin-skinned fibers showed that 4-CmC induced a dose-dependent caffeine-like transient contractile response in edl and soleus due to an activation of the ryanodine receptor. Both types of skeletal muscles showed a 10 to 20 times lower 4-CmC threshold concentration and EC(50) value (concentration providing 50% of the maximal 4-CmC contracture) than for caffeine. The results indicate that edl is more sensitive than soleus to 4-CmC and that this difference in sensitivity is more marked than with caffeine. Furthermore, an increase in cytosolic Ca(2+) activity induced a more marked shift of dose-response curves toward lower concentrations for 4-CmC than caffeine. Experiments conducted on Triton X-100-skinned fibers showed that in both muscles, 4-CmC decreased in a dose-dependent manner the Ca(2+)-activated force of contractile apparatus, particularly in edl. Furthermore, the tension pCa curves indicated that 4-CmC induced a dose-dependent sensitizing (soleus) or desensitizing (edl) effect on the Ca(2+) sensitivity of myofibrils. These results indicate that edl and soleus contractile responses can be discriminated with 4-CmC instead of caffeine and that care must be taken in interpreting results because muscular pathology could be due in part to an increase in intracellular Ca(2+).

Ciliary neurotrophic factor prevents unweighting-induced functional changes in rat soleus muscle.

The purpose of the present work was to see whether changes in rat soleus characteristics due to 3 wk of hindlimb suspension could be modified by ciliary neurotrophic factor (CNTF) treatment. Throughout the tail suspension period, the cytokine was delivered by means of an osmotic pump (flow rate 16 microg. kg(-1). h(-1)) implanted under the hindlimb skin. In contrast to extensor digitorum longus, CNTF treatment was able to reduce unweighting-induced atrophy in the soleus. Twitch and 146 mM potassium (K) tensions, measured in small bundles of unloaded soleus, decreased by 48 and 40%, respectively. Moreover, the time to peak tension and the time constant of relaxation of the twitch were 48 and 54% faster, respectively, in unloaded soleus than in normal muscle. On the contrary, twitch and 146 mM K contracture generated in CNTF-treated unloaded and normal soleus were not different. CNTF receptor-alpha mRNA expression increased in extensor digitorum longus and soleus unloaded nontreated muscles but was similar in CNTF-treated unloaded muscles. The present results demonstrate that exogenously provided CNTF could prevent functional changes occurring in soleus innervated muscle subject to unweighting.

Rapid cooling-induced contractures in rat skinned skeletal muscle fibres originate from sarcoplasmic reticulum Ca2+ release through ryanodine and inositol trisphosphate receptors.

Previous reports have shown that cooling striated muscles induces contractile responses that are related to Ca2+ release from the sarcoplasmic reticulum. However, the effect of cooling has generally been studied in the presence of pharmacological agents that potentiate rapid cooling-induced contractures. The present study shows that in saponin-skinned rat skeletal muscle preparations, a drop in temperature from 22 degrees C to 2 degrees C per se induces a contracture which relaxes on return to 22 degrees C. In fast-twitch fibres, rapid cooling-induced contractures are fully blocked by ryanodine, an inhibitor of ryanodine receptors. By contrast, in slow-twitch fibres, ryanodine partially inhibits the rapid cooling-induced contractile response, leaving a residual tension that dissipates after application of inositol 1,4,5-trisphosphate (InsP3). At low concentrations, heparin, an inhibitor of InsP3 receptors, decreases rapid cooling-induced contractures in both types of muscle. The present results suggest that in skeletal muscle, rapid cooling-induced contractures are due to both ryanodine-sensitive and InsP3-sensitive Ca2+ release from the sarcoplasmic reticulum.

Inositol 1,4,5-trisphosphate-sensitive Ca2+ release in rat fast- and slow-twitch skinned muscle fibres.

Inositol 1,4,5-trisphosphate (InsP3), an intracellular messenger, induces Ca2+ release in various types of cells, particularly smooth muscle cells. Its role in skeletal muscle, however, is controversial. The present study shows that the application of InsP3 to rat slow- and fast-twitch saponin-skinned fibres induced contractile responses that were not related to an effect of InsP3 on the properties of the contractile proteins. The amplitude of the contractures was dependent upon the Ca(2+)-loading period, and was larger in slow- than in fast-twitch muscle. In both types of skeletal muscle, these responses, unlike caffeine contractures, were not inhibited by ryanodine (100 microM), but were abolished by heparin (20 micrograms.ml-1). In soleus muscle, the concentration of heparin required to inhibit the response by 50% (IC50) was 5.7 micrograms.ml-1, a similar value to that obtained previously in smooth muscle. Furthermore, the results show that in slow-twitch muscle, the InsP3 contractures have a "bell-shaped" dependency on the intracellular Ca2+ concentration. These results show that InsP3 receptors should be present in skeletal muscle. Thus, it is possible that InsP3 participates in the regulation of sarcoplasmic reticulum Ca2+ release in skeletal muscle, particularly in slow-twitch fibres.

Na(+)-Ca2+ exchange induces low Na+ contracture in frog skeletal muscle fibers after partial inhibition of sarcoplasmic reticulum Ca(2+)-ATPase.

Contractile responses due to reduction in external sodium concentration ([Na+]o) were investigated in twitch skeletal muscle fibers of frog semitendinosus. Experiments were conducted after partial inhibition of sarcoplasmic reticulum Ca(2+)-ATPase by cyclopiazonic acid (CPA). In the absence of CPA, Na+ withdrawal failed to produce any change in resting tension. In the presence of CPA (2-10 microM), [Na+]o reduction induced a transient contracture without a significant change in the resting membrane potential. The amplitude of the contracture displayed a step dependence on [Na+]o, was increased by K(+)-free medium and was prevented in Ca(2+)-free medium. This contracture was inhibited by various blockers of the Na(+)-Ca2+ exchange but was little affected by inhibitors of sarcolemmal Ca(2+)-ATPase or mitochondria. When sarcoplasmic reticulum function was impaired, low-Na+ solutions caused no contracture. These results provide evidence that skeletal muscle fibers possess a functional Na(+)-Ca2+ exchange which can mediate sufficient Ca2+ entry to activate contraction by triggering Ca2+ release from sarcoplasmic reticulum when the sodium electrochemical gradient is reduced, and sarcoplasmic reticulum Ca(2+)-ATPase is partially inhibited. This indicates that when the sarcoplasmic reticulum Ca(2+)-ATPase is working (no CPA), Ca2+ fluxes produced by the exchanger are buffered by the sarcoplasmic reticulum. Thus the Na(+)-Ca2+ exchange may be one of the factors determining sarcoplasmic reticulum Ca2+ content and thence the magnitude of the release of Ca2+ from the sarcoplasmic reticulum.

Changes in voltage activation of contraction in frog skeletal muscle fibres as a result of sarcoplasmic reticulum Ca2+-ATPase activity.

The effects of cyclopiazonic acid, a specific sarcoplasmic reticulum Ca2+-ATPase inhibitor, on isometric tension were studied in response to prolonged steady-state depolarization induced by a rapid change in extracellular potassium concentration (potassium contractures) in frog semitendinosus muscle fibres. Cyclopiazonic acid (1-10 microM) enhanced the amplitude and time-course of relaxation of 146 mM potassium contracture. In the presence of cyclopiazonic acid 0.5 microM, the relationship between the amplitude of potassium contractures and the membrane potential shifted to more negative potentials, whereas the steady-state inactivation curve was unchanged. These observations suggest that cyclopiazonic acid has no effect on voltage sensors. The difference between potassium contractures in the absence and presence of cyclopiazonic acid in skeletal muscle fibres implies that the amplitude and slow relaxation of tension during prolonged steady-state depolarization may be expected to depend not only on inactivation of the process regulating calcium release from the sarcoplasmic reticulum but also on the ability of the sarcoplasmic reticulum to pump calcium.

Sarcoplasmic reticulum Ca(2+) release by 4-chloro-m-cresol (4-CmC) in intact and chemically skinned ferret cardiac ventricular fibers.

The purpose of this study was to determine whether 4-chloro-m-cresol (4-CmC) could generate caffeine-like responses in ferret cardiac muscle. The concentration dependence of 4-CmC-mediated release of Ca(2+) from the sarcoplasmic reticulum was studied in intact cardiac trabeculae and saponin-skinned fibers in which the sarcoplasmic reticulum was loaded with Ca(2+). In intact and saponin-skinned preparations isolated from right ventricle, the effect of 4-CmC on sarcoplasmic reticulum Ca(2+) content was estimated by analysis of caffeine contracture after application of chlorocresol. In addition, the effects of 4-CmC on maximal Ca(2+)-activated tension and the Ca(2+) sensitivity of myofibrils were analyzed by using Triton-skinned cardiac fibers. The results show that 4-CmC generates a contractile response in saponin-skinned but not intact fibers. The sarcoplasmic reticulum is implicated in the 4-CmC response; more precisely, in Ca(2+) release via the ryanodine receptor. Moreover, 4-CmC, like caffeine, has effects on maximal Ca(2+)-activated tension and the Ca(2+) sensitivity of myofibrils.

Measurement of sarcomere length during fast contraction of muscle fibers by digital image analysis.

A low-cost, high-resolution (spatial and temporal) image analysis system was developed to measure sarcomere length (Sl) during fast twitch of isolated striated muscle fibers at different temperatures. Fiber images were examined during twitch with an imaging rate of 220 Hz. To increase temporal resolution beyond 220 Hz, consecutive temporally shifted image sequences (N sequences) were acquired. Individual or average Sl was directly measured from a horizontal profile without spatial-frequency assessment. Measurement precision (E) was determined and expressed as: E(%) = 100xPs/(IsxSl), where Ps is the pixel size and Is the involved sarcomere number. At 18 degrees C during isometric twitch, Sls were measured with 220 Hz temporal and 0.2% spatial resolutions. Sl shortened in the central region (0.21+/-0.12 microm) as tension developed, reaching a maximal shortening of 8.09 + 2.05% (at rest, Sl = 2.59+/-0.05 microm, n = 4) in 32.5+/-1.96 ms. At 30 degrees C, Sl variations were examined with 880 Hz temporal resolution, in which case maximal S1 shortening was reached in 15.74+/-1.99 ms, and then decreased to 5.19+/-1.97% (at rest, S1 = 2.6+/-0.06 microm). The twitch tension developed by the whole fiber was recorded and compared with sarcomere length behavior. Sarcomere length variations in the central region were representative of overall developed tensions at 18 and 30 degrees C.

Changes in CNTF receptor alpha expression in rat skeletal muscle during the recovery period after hindlimb suspension.

This study investigated whether modifications in the contractile function of rat muscle after 3 weeks of hindlimb suspension followed by different recovery periods were associated with changes in ciliary neurotrophic factor (CNTF) receptor alpha. CNTF receptor alpha was stronger in control extensor digitorum longus (EDL) than soleus. CNTF receptor alpha was increased in unweighted as compared to control soleus muscle, while no significant changes occurred in EDL muscle. Furthermore, CNTF level was not significantly modified in adult sciatic nerve.

Cyclopiazonic acid-induced changes in the contraction and Ca2+ transient of frog fast-twitch skeletal muscle.

The effects of cyclopiazonic acid (CPA) were investigated on isolated skeletal muscle fibers of frog semitendinosus muscle. CPA (0.5-10 microM) enhanced isometric twitch but produced little change in resting tension. At higher concentrations (10-50 microM), CPA depressed twitch and induced sustained contracture without affecting resting and action potentials. In Triton-skinned fibers, CPA had no significant effect on myofibrillar Ca2+ sensitivity but decreased maximal activated force at concentrations > 5 microM. In intact cells loaded with the Ca2+ fluorescence indicator indo 1, CPA (2 microM) induced an increase in Ca(2+)-transient amplitude (10 +/- 2.5%), which was associated with an increase in time to peak and in the time constant of decay. Consequently, peak force was increased by 35 +/- 4%, and both time to peak and the time constant of relaxation were prolonged. It is concluded that CPA effects, at a concentration of up to 2 microM, were associated with specific inhibition of sarcoplasmic reticulum Ca(2+)-adenosinetriphosphatase in intact skeletal muscle and that inhibition of the pump directly affected the handling of intracellular Ca2+ and force production.

Potential targets for skeletal muscle impairment by hypogravity: basic characterization of resting ionic conductances and mechanical threshold of rat fast- and slow-twitch muscle fibers.

Prolonged hypogravity such as during space flights affects skeletal muscle function by inducing postural changes as well as reduced muscle strength and locomotion capacity. Also in rats, space flight as well as useful models of groundbased hypogravity induce marked atrophy in the slow-twitch soleus (SOL) muscle as opposed to slight or none in the fast-twitch ones such as extensor digitorum longus (EDL). Biochemical and histological studies on hindlimb suspended animals, showed a hypogravity-induced impairment of muscle function involving the transition of slow-twitch muscle type, responsible for postural control, toward the fast-twitch phenotype by modification of excitation-contraction pattern. In slow muscles of rats, hindlimb suspension induced upregulation of the fast isoform of myosin heavy-chain and increased expression of fast Ca2+ pump mRNA and protein, which is consistent with the increased Ca(2+)-dependent ATPase activity and the speeding of muscle relaxation, typical of fast muscles. Little is known about the modifications induced by hypogravity in the sarcolemmal ion channels function, which controls the pattern of muscle excitability and contractility. The normally high resting chloride conductance, which is required for the electrical stabilization of mammalian muscle fibers, may be a target of hypogravity modifications since a pharmacological block of this parameter determines, though an increase of excitability, the transition of the fast-twitch muscle phenotype toward the slow one either in adult or in developing rats. Hypogravity also induced increased expression of dihydropyridine receptors in soleus muscle, that are normally lower than that found in the fast ones. In this study, we characterized the electrical and contractile properties of rat extensor digitorum longus (EDL) and slow-twitch soleus SOL muscles fibers at the aim to better understand the molecular mechanisms leading to fiber transition.