Fast-twitch Extensor Digitorum Longus Muscles Research Articles

Effects of thyroid hormone receptor gene disruption on myosin isoform expression in mouse skeletal muscles.

Skeletal muscle is known to be a target for the active metabolite of thyroid hormone, i.e., 3,5,3'-triiodothyronine (T(3)). T(3) acts by repressing or activating genes coding for different myosin heavy chain (MHC) isoforms via T(3) receptors (TRs). The diverse function of T(3) is presumed to be mediated by TR-alpha(1) and TR-beta, but the function of specific TRs in regulating MHC isoform expression has remained undefined. In this study, TR-deficient mice were used to expand our knowledge of the mechanisms by which T(3) regulates the expression of specific MHC isoforms via distinct TRs. In fast-twitch extensor digitorum longus (EDL) muscle, TR-alpha(1)-, TR-beta-, or TR-alpha(1)beta-deficient mice showed a small but statistically significant decrease (P < 0.05) of type IIB MHC content and an increased number of type I fibers. In the slow-twitch soleus, the beta/slow MHC (type I) isoform was significantly (P < 0. 001) upregulated in the TR-deficient mice, but this effect was highly dependent on the type of receptor deleted. The lack of TR-beta had no significant effect on the expression of MHC isoforms. An increase (P < 0.05) of type I MHC was observed in the TR-alpha(1)-deficient muscle. A dramatic overexpression (P < 0.001) of the slow type I MHC and a corresponding downregulation of the fast type IIA MHC (P < 0.001) was observed in TR-alpha(1)beta-deficient mice. The muscle- and fiber-specific differences in MHC isoform expression in the TR-alpha(1)beta-deficient mice resembled the MHC isoform transitions reported in hypothyroid animals, i.e., a mild MHC transition in the EDL, a dramatic but not complete upregulation of the beta/slow MHC isoform in the soleus, and a variable response to TR deficiency in different soleus muscle fibers. Thus the consequences on muscle are similar in the absence of thyroid hormone or absence of thyroid hormone receptors, indicating that TR-alpha(1) and TR-beta together mediate the known actions of T(3). However, it remains unknown how thyroid hormone exerts muscle- and muscle fiber-specific effects in its action. Finally, although developmental MHC transitions were not studied specifically in this study, the absence of embryonic and fetal MHC isoforms in the TR-deficient mice indicates that ultimately the transition to the adult MHC isoforms is not solely mediated by TRs.

Sublytic complement attack increases intracellular sodium in rat skeletal muscle.

Although excessive complement activation and deranged sodium homeostasis in skeletal muscle are characteristic in sepsis, their relationship has not been examined. This study was designed to determine if sublytic complement activation can directly mediate changes in myocellular sodium content. Fast-twitch extensor digitorum longus muscles were freshly isolated from infant rats. Unsensitized muscles were incubated at 30 degrees C for 60 min in the media containing 10% human or rat serum under conditions of no complement activation, activation by zymosan, inactivation by heat, C7 or C9 deficiency, selective inhibition of complement pathway, and inhibition of Na(+)-K(+) ATPase by ouabain. Intracellular sodium ([Na(+)](i)) and potassium ([K(+)](i)) contents of the muscles, myocellular ATP, and LDH release from the muscles were then determined. Normal human serum significantly increased [Na(+)](i) and the [Na(+)](i)/[K(+)](i) ratio in the muscles as well as zymosan-activated serum. Heat inactivation, C7 deficiency, and inhibition of the alternative pathway completely abolished the cationic changes. Average LDH release was identical in all groups and less than 6%. Complement activation did not impair ouabain-sensitive Na(+)-K(+) ATPase activity in the muscles or alter myocellular ATP. Thus, the observed alterations are not likely due to dysfunction of Na(+)-K(+) pump or depletion of myocellular energy. Instead, alterations in [Na(+)](i) were dependent upon the amount of C9 added to C9-deficient serum, which suggests that the alterations are likely dependent on transmembrane pores created by membrane attack complexes (MAC). Sublytic amounts of MAC formed as a result of complement activation can directly alter [Na(+)](i) in ex vivo skeletal muscle.

NNOS and eNOS modulate cGMP formation and vascular response in contracting fast-twitch skeletal muscle

Nitric oxide (NO) from Ca(2+)-dependent neuronal nitric oxide synthase (nNOS) in skeletal muscle fibers may modulate vascular tone by a cGMP-dependent pathway similar to NO derived from NOS in endothelial cells (eNOS). In isolated fast-twitch extensor digitorum longus (EDL) muscles from control mice, cGMP formation increased approximately 166% with electrical stimulation (30 Hz, 15 s). cGMP levels were not altered in slow-twitch soleus muscles. The NOS inhibitor N(omega)-nitro-l-arginine abolished the contraction-induced increase in cGMP content in EDL muscles, and the NO donor sodium nitroprusside (SNP) increased cGMP content approximately 167% in noncontracting EDL muscles. SNP treatment but not electrical stimulation increased cGMP formation in muscles from nNOS(-/-) mice. cGMP formation in control and stimulated EDL muscles from eNOS(-/-) mice was less than that obtained with similarly treated muscles from control mice. Arteriolar relaxation in contracting fast-twitch mouse cremaster muscle was attenuated in muscles from mice lacking either nNOS or eNOS. These findings suggest that increases in cGMP and NO-dependent vascular relaxation in contracting fast-twitch skeletal muscle may require both nNOS and eNOS.

Hydrogen peroxide modulates Ca2+-activation of single permeabilized fibres from fast- and slow-twitch skeletal muscles of rats.

We examined the effects of redox modulation on single membrane-permeabilized fibre segments from the fast-twitch extensor digitorum longus (EDL) and slow-twitch soleus muscles of adult rats to determine whether the contractile apparatus was the redox target responsible for the increased contractility of muscles exposed to low concentrations of H2O2. The effects of H2O2 on maximum Ca2+-activated force were dose-dependent with 30 min exposure to 5 mM H2O2 causing a progressive decrease by 22+/-3 and 13+/-2% in soleus and EDL permeabilized muscle fibres, respectively. Lower concentrations of exogenous H2O2 (100 microM and 1 mM) had no effect on maximum Ca2+-activated force. Subsequent exposure to the reductant dithiothreitol (DTT, 10 mM, 10 min) fully reversed the H2O2-induced depression of force in EDL, but not in soleus muscle fibres. Incubation with DTT alone for 10 min did not alter Ca2+-activated force in either soleus or EDL muscle fibres. The sensitivity of the contractile filaments to Ca2+ (pCa50) was not altered by exposure to any concentration of exogenous H2O2. However, all concentrations of H2O2 diminished the Hill coefficient in permeabilized fibres from the EDL muscle, indicating that the cooperativity of Ca2+ binding to troponin is altered. H2O2 (5 mM) did not affect rigor force, which indicates that the number of crossbridges participating in contraction was not reduced. In conclusion, H2O2 may reduce the maximum Ca2+ activated force production in skinned muscle fibres by decreasing the force per crossbridge.

Muscle-Specific Up-Regulation of Rat UCP3 mRNA Expression by Long-Term Hindlimb Unloading

The effect of long-term hindlimb unloading (2 or 5 week) on the expression of uncoupling protein-3 (UCP3) gene was investigated in rat skeletal muscles. The interaction of hindlimb unloading and thyroid status was also investigated at 2 weeks. Whatever the duration, mechanical unloading induced a similar increase in UCP3 mRNA relative abundance in the slow-twitch soleus (SOL) muscle (+80%, P < 0.05), whereas no effect was observed in the fast-twitch extensor digitorum longus (EDL) muscle. Hypothyroidism down-regulated while hyperthyroidism up-regulated UCP3 mRNA relative abundance in both SOL and EDL muscles, but thyroid status did not prevent the up-regulation of UCP3 induced by 2 weeks of suspension. These data therefore indicate for the first time that long-term hindlimb unloading up-regulates muscle UCP3 gene expression in a muscle-specific manner which is independent of thyroid status.

Adaptation of mouse skeletal muscle to a novel functional overload test: changes in myosin heavy chains and SERCA and physiological consequences.

We have used a new approach to study the effects of overload on skeletal muscle phenotype in mice. The method used avoids any traumatising contact with muscles and the inflammatory reaction that this may provoke. Blocks of lead embedded in silicone were inserted under the skin of the lower part of the back. After 1 month, a 17% hypertrophy was found to have occurred in the tonic soleus muscle, but no change was observed in the fast-twitch extensor digitorum longus (EDL) muscle. The main effects on the contractile properties of the soleus muscle were a decrease in the tetanic relaxation rate and a reduction in the maximal velocity of shortening. Immunohistological analysis of the soleus muscles revealed an increase in the proportion of fibres that express myosin heavy chain (MHC) 1, from 54.2% to 73.9%, with a reduction in the proportion of MHC2a-positive fibres, from 45.8% to 30.2%. These changes were accompanied by an increase in the proportion of fibres that express the slow type of sarcoplasmic reticulum calcium pump (SERCA2a), from 61.8% to 84.7%. In EDL muscles, overload induced only minor changes. Thus, this method of overload affected the soleus muscle in particular. The observed changes in the control of muscle contraction were significantly larger than the changes in typical myofibrillar properties that were observed. These results indicate that there is a temporal dissociation between the relative expression of MHCs and SERCAs.

Different metabolic adaptation of heart and skeletal muscles to moderate-intensity treadmill training in the rat.

The effect was investigated of treadmill training of moderate intensity on the fatty acid-binding protein (FABP) content in relation to parameters of oxidative and glycolytic metabolism. To this end, the cytoplasmic FABP content and the activity of beta-hydroxyacyl-coenzyme A dehydrogenase (HAD), citrate synthase (CS), and 6-phosphofructokinase (PFK) were measured in heart, fast-twitch extensor digitorum longus (EDL) and slow-twitch soleus muscles (SOL) of male Wistar rats. To investigate the influence of the amount of training (defined as the product of exercise duration, intensity and frequency), two training groups were created that differed in training frequency (HF, high frequency 5 days x week(-1), n = 9; LF, low frequency 2 days x week(-1), n = 9; the exercise being 20 m x min(-1) for 2 h with no gradient, over 6 weeks) and compared with SC, sedentary controls (n = 7). In heart muscle, the cytoplasmic FABP content was 34% higher in HF than in SC but was the same as in LF. The CS and HAD activities were no different in the three groups, suggesting that the capacity to oxidize fatty acids (FA) was not affected by training. The PFK activity was higher (43%) in HF, suggesting a shift towards carbohydrate utilization. The FABP content and HAD activity did not change in SOL and EDL after training whereas the CS activity increased (27%) in SOL and decreased (21%) in EDL in both training groups. In addition, PFK activity in EDL was much higher (113%) in the HF than in SC group. The HF training was associated with a fine-tuning of FA availability and use in heart muscle, and with a more efficient energy production. It is suggested therefore that cytoplasmic FABP could be an early marker of muscle adaptation to training in heart but not in skeletal muscle. The training reinforced the metabolic profile of the skeletal muscles, in particular that of the fast-twitch glycolytic muscle. We concluded that a large amount of training is needed when the effect on both oxidative and glycolytic parameters is to be studied.

Sepsis is associated with increased ubiquitinconjugating enzyme E214k mRNA in skeletal muscle.

Previous studies provided evidence that sepsis is associated with increased ubiquitin-proteasome-dependent protein breakdown in skeletal muscle. The 14-kDa ubiquitin-conjugating enzyme (E214k) has been proposed to be a key regulator of the ubiquitin proteolytic pathway. We tested the hypothesis that E214k message and protein levels are increased in skeletal muscle during sepsis. Sepsis was induced in rats by cecal ligation and puncture (CLP). Control rats were sham operated. E214k mRNA and protein levels were quantitated after Northern and Western blot analysis, respectively, 16 h after CLP or sham operation. Sepsis resulted in a 70% increase in the 1. 2-kb E214k transcript in the fast-twitch extensor digitorum longus muscle, whereas no changes were seen in the slow-twitch soleus muscle. E214k protein levels were not influenced by sepsis in any of the muscles studied. Although the changes in the expression of the E214k 1.2-kb transcript paralleled the differential effect of sepsis on protein breakdown in fast- and slow-twitch muscle, the potential role of E214k in the regulation of sepsis-induced muscle proteolysis needs to be interpreted with caution, because the results demonstrated that increased message levels were not associated with increased E214k protein levels.

Quantification of thyroid hormone receptor isoforms, 9-cis retinoic acid receptor gamma, and nuclear receptor co-repressor by reverse-transcriptase PCR in maturing and adult skeletal muscles of rat.

Quantitative competitive reverse-transcriptase polymerase chain reaction (qcRT-PCR) was established for determining absolute molecule numbers of the thyroid hormone receptor (T3R) isoforms T3Ralpha1, T3Ralpha2, T3Rbeta1, and the 9-cis retinoic acid receptor gamma (RXRgamma) in developing and adult fast-twitch extensor digitorum longus (EDL) and slow-twitch soleus (SOL) muscles of rat. Expression levels of the nuclear receptor co-repressor (NCoR) were measured in the same muscles because responses to thyroid hormones during muscle maturation might not only depend on the expression levels of the various receptors but might also be modulated by changes in the expression of NCoR. The qcRT-PCR method was based on the addition of known amounts of homologous competitor RNAs to the reverse transcriptase (RT) reaction. We show that all nuclear receptors under study were expressed in fast and slow muscles. Transcript numbers of T3Rbeta1, which was the most abundant isoform, were higher in SOL than in EDL during all developmental stages. The mRNAs for T3Ralpha1, T3Ralpha2, RXRgamma and the NCoR displayed molecule numbers in similar ranges, but were differentially expressed. T3Ralpha1 mRNA increased in SOL during postnatal development, while T3Ralpha2 mRNA initially decreased, then increased to adult levels. Conversely, pronounced decreases were observed for T3Ralpha1 (10-fold) and T3Ralpha2 (28-fold) mRNAs in the EDL muscle during postnatal maturation. RXRgamma mRNA was 10-fold downregulated during EDL maturation, but unaltered in maturing SOL. NCoR transcript number displayed only minor changes in both muscles.

Role of extracellular [Ca2+] in fatigue of isolated mammalian skeletal muscle.

The possible role of altered extracellular Ca2+ concentration ([Ca2+]o) in skeletal muscle fatigue was tested on isolated slow-twitch soleus and fast-twitch extensor digitorum longus muscles of the mouse. The following findings were made. 1) A change from the control solution (1.3 mM [Ca2+]o) to 10 mM [Ca2+]o, or to nominally Ca2+-free solutions, had little effect on tetanic force in nonfatigued muscle. 2) Almost complete restoration of tetanic force was induced by 10 mM [Ca2+]o in severely K+-depressed muscle (extracellular K+ concentration of 10-12 mM). This effect was attributed to a 5-mV reversal of the K+-induced depolarization and subsequent restoration of ability to generate action potentials (inferred by using the twitch force-stimulation strength relationship). 3) Tetanic force depressed by lowered extracellular Na+ concentration (40 mM) was further reduced with 10 mM [Ca2+]o. 4) Tetanic force loss at elevated extracellular K+ concentration (8 mM) and lowered extracellular Na+ concentration (100 mM) was partially reversed with 10 mM [Ca2+]o or markedly exacerbated with low [Ca2+]o. 5) Fatigue induced by using repeated tetani in soleus was attenuated at 10 mM [Ca2+]o (due to increased resting and evoked forces) and exacerbated at low [Ca2+]o. These combined results suggest, first, that raised [Ca2+]o protects against fatigue rather than inducing it and, second, that a considerable depletion of [Ca2+]o in the transverse tubules may contribute to fatigue.

Myosin Heavy Chain Expression in Skeletal Muscle Autografts under Neural or Aneural Conditions

Background.Our purpose was to investigate (1) the heterogeneity of satellite cells derived from adult fast-twitch and slow-twitch skeletal muscles, (2) the influence of innervation on muscle regeneration, and (3) the differences between developmental myoblasts and satellite cells with regard to myosin heavy chain (MHC) expression.Materials and methods.Autografts under neural (nerve-intact graft;brief denervation interval) or aneural (aneural graft;prolonged denervation interval) conditions of the fast-twitch extensor digitorum longus (EDL) muscle or the slow-twitch soleus muscle were performed in adult rat hindlimbs. MHC expression during skeletal muscle regeneration was determined sequentially using immunocytochemistry.Results.After grafting, most muscle fibers in the EDL and soleus underwent ischemic degeneration and regeneration; at the periphery of each muscle, a few adult fibers survived. All regenerating fibers initially expressed embryonic/fetal (developmental) MHC alone, and subsequently both developmental and fast MHC. During the first week, no expression of slow MHC was observed in regenerating fibers in either the EDL or the soleus. In nerve-intact grafts, regenerating fibers expressed slow MHC as early as the second week; under aneural conditions, no regenerating fibers expressed slow MHC even 4 weeks after grafting. On the other hand, some persisting fibers in aneural grafts could maintain expression of slow MHC 4 weeks after grafting; other fibers underwent MHC transformation induced by denervation. No significant difference in MHC expression during regeneration was observed for slow compared with fast muscles, under either neural or aneural condition.Conclusions.These data suggest that regenerating adult skeletal muscle fibers, derived only from satellite cells, cannot express slow MHC without motor innervation, and that persisting muscle fibers, derived from both myoblasts in fetal development and satellite cells, may be intrinsically distinct from regenerating fibers. Satellite cells derived from slow and from fast muscles may be a single, homogenous population and may be the same population as fetal (secondary) myoblasts with regard to MHC expression.

Decay Of Ca2+ and Force Transients in Fast- and Slow-Twitch Skeletal Muscles from the Rat, Mouse and Etruscan Shrew

Isometric single-twitch force and intracellular Ca2+ transients were recorded simultaneously, using fura-2, from slow- and fast-twitch muscle fibres of the rat, mouse and Etruscan shrew Suncus etruscus. In the slow-twitch rat soleus, force half-relaxation time was three times longer than the 50% decay time of the fura-2 signal. In contrast, in the fast-twitch extensor digitorum longus muscles of all three animals, muscle relaxation preceded Ca2+ decay. It is proposed that this surprising property of fast-twitch muscles is due to their pCa-tension curve, which is shifted to the right compared with that of slow-twitch muscle.

Use of DNA injection for identification of slow nerve-dependent regions of the MLC2sgene

It has been well established that expression of slow contractile protein genes in skeletal muscle is regulated, in part, by activity from slow motoneurons. However, very little is understood about the mechanism by which neural activity regulates transcription of slow isoform genes. The purpose of this investigation was first to more fully define the in vivo DNA injection technique for use in both fast-twitch and slow-twitch muscles and second to use the injection technique for the identification of slow nerve-dependent regions of the myosin light chain 2 slow (MLC2s) gene. Initial experiments determined that the same amount of plasmid DNA was taken up by both the slow-twitch soleus and fast-twitch extensor digitorum longus (EDL) muscles and that injection of from 0.5 to 10 micrograms DNA/muscle is ideal for analysis of promoter activity during regeneration. This technique was subsequently used to identify that the region from -800 to +12 base pairs of MLC2s gene directed approximately 100 times higher activity in the innervated soleus than in innervated EDL, denervated soleus, or denervated EDL muscles. Placing the introns upstream of either the MLC2s or SV40 promoter increased expression 5- and 2.7-fold, respectively, in innervated soleus but not in innervated EDL, denervated soleus, or denervated EDL muscles. These results demonstrate that 1) in vivo DNA injection is a sensitive assay for promoter analysis in both fast-twitch and slow-twitch skeletal muscles and 2) both 5' flanking and intronic regions of the MLC2s gene can independently and synergistically direct slow nerve-dependent transcription in vivo.

Energetics of lengthening in mouse and toad skeletal muscles.

1. The energetics of lengthening were studied in amphibian and mammalian skeletal muscle. The aims were to determine whether energy absorption during stretch is a general property of skeletal muscle and to investigate the influence of lengthening velocity on energy absorption. 2. Experiments were performed in vitro (21 degrees C) using bundles of muscle fibres from fast-twitch extensor digitorum longus and slow-twitch soleus muscles of the mouse and tibialis anterior muscles of a toad, Bufo marinus. Initial heat production and mechanical work done on muscles were measured during isovelocity lengthening. Enthalpy output during lengthening was calculated as the difference between the amount of heat produced and the work done. 3. For all three muscle types, more energy was put into muscles as work than was produced as heat. Thus, part of the energy put into muscles to stretch them must have been absorbed. 4. For all three muscle types, the amount of energy absorbed was constant at velocities exceeding approximately 0.5 Vmax (Vmax is the maximum shortening velocity), but was significantly lower at slow velocities of lengthening. The same amount of energy was absorbed by all three muscles when lengthened at > or = 0.5 Vmax. 5. It was concluded that absorption of energy during lengthening occurs in mammalian as well as amphibian muscle and that lengthening velocity has only a small effect on the amount of energy absorbed.

Muscle-specific transgenic complementation of GLUT4-deficient mice. Effects on glucose but not lipid metabolism.

We have taken the approach of introducing the muscle-specific myosin light chain (MLC)-GLUT4 transgene into the GLUT4-null background to assess the relative role of muscle and adipose tissue GLUT4 in the etiology of the GLUT4-null phenotype. The resulting MLC-GLUT4-null mice express GLUT4 predominantly in the fast-twitch extensor digitorum longus (EDL) muscle. GLUT4 is nearly absent in female white adipose tissue (WAT) and slow-twitch soleus muscle of both sexes of MLC-GLUT4-null mice. GLUT4 content in male MLC-GLUT4-null WAT is 20% of that in control mice. In transgenically complemented EDL muscle, 2-deoxyglucose (2-DOG) uptake was restored to normal (male) or above normal (female) levels. In contrast, 2-DOG uptake in slow-twitch soleus muscle of MLC-GLUT4-null mice was not normalized. With the normalization of glucose uptake in fast-twitch skeletal muscle, whole body insulin action was restored in MLC-GLUT4-null mice, as shown by the results of the insulin tolerance test. These results demonstrate that skeletal muscle GLUT4 is a major regulator of skeletal muscle and whole body glucose metabolism. Despite normal skeletal muscle glucose uptake and insulin action, the MLC-GLUT4-null mice exhibited decreased adipose tissue deposits, adipocyte size, and fed plasma FFA levels that are characteristic of GLUT4-null mice. Together these results indicate that the defects in skeletal muscle and whole body glucose metabolism and adipose tissue in GLUT4-null mice arise independently.

Different effects of raised [K+]o on membrane potential and contraction in mouse fast- and slow-twitch muscle.

Increasing extracellular K+ concentration ([K+]o) from 4 to 7-14 mM reduced both tetanic force and resting membrane potential (Em) in isolated slow-twitch soleus and fast-twitch extensor digitorum longus (EDL) muscles of the mouse. The tetanic force-[K+]o relationships showed a greater force loss over 8-11 mM [K+]o in soleus than EDL, mainly because the Em was 2-3 mV less negative at each [K+]o in soleus. The tetanic force-resting Em relationships show that force was reduced in two phases: phase 1 (Em < -60 mV), a 20% force decline in which the relationships superimposed in soleus and EDL, and phase 2 (Em -60 to -55 mV), a marked force decline that was steeper in EDL than soleus. Additionally in phase 2, longer stimulation pulses restored tetanic force; the twitch force-stimulation strength relationship was shifted toward higher voltages; caffeine, a myoplasmic Ca2+ concentration elevator, increased maximum force; and twitch force fell abruptly. We suggest that 1) the K(+)-depressed force is due to reduced Ca2+ release resulting from an altered action potential profile (phase 1) and inexcitable fibers due to an increased action potential threshold (phase 2), and 2) K+ contributes to fatigue in both fast- and slow-twitch muscle when it causes depolarization to about -60 mV.

An investigation into the role of rat skeletal muscle as a site for xenobiotic metabolism using microsomes and isolated cells

1. The role of skeletal muscle microsomes as a site of extrahepatic xenobiotic metabolism using n-hexane as a model substrate was investigated. The observed cytochrome P450-dependent metabolism was compared with that found with liver, and brain microsomal fractions. 2. Rat skeletal muscle microsomes metabolised n-hexane to 1-, 2- and 3-hexanol at rates 40-300 times lower than observed with rat liver microsomes. 3. Fast-twitch extensor digitorum longus muscle (EDL) microsomes had twice as much n-hexane hydroxylase activity as the slow-twitch soleus and furthermore the EDL microsomes produced 2-hexanol, a bioactivation product of n-hexane, as a major metabolite. 4. Metabolism of hexane to 1-, 2- and 3-hexanol and 2-hexanon was demonstrated in cultured rat myoblasts. 5. Ethoxyresorufin and pentoxyresorufin O-dealkylation were not detected in either muscle microsomes or myoblasts although immunocytochemical localisation studies were suggestive of the presence of cytochrome P-450. 6. In conclusion, rat skeletal muscle has a low level of xenobiotic metabolism activity. The relevance to neuromuscular toxicity of n-hexane is discussed.

Gender‐related differences in the regulatory influence of thyroid hormone on the expression of myosin isoforms in young and old rats

The effects of 4 weeks of thyroid hormone (3.5.3'-triiodothyronine. T3) treatment on the expression of myosin heavy chain (MyHC) isoforms were examined in young (3-6 months; and old (20-24 months) female rats, and compared with those in age-matched male rats (Larsson et al. 1995). In control rats ageing was associated with a type IIA to I MyHC isoform switching in the slow-twitch soleus and a type IIB to IIX MyHC isoform switching in the fast-twitch extensor digitorum longus muscle (EDL). Gender- and muscle-specific differences were observed in the regulation of MyHC isoforms by T3. In the soleus, dramatic downregulation of the type I and upregulation of the type IIA MyHC isoform were observed in both females and males, but upregulation of the IIX MyHC isoform was observed only in male rats. In EDL, T3 treatment had no significant influence on the MyHC isoform composition in the males irrespective of the age of the animal. In the females, on the other hand, T3 treatment resulted in a significant MyHC transformation from IIA to IIB, probably via IIX myosin, in spite of the fact that type IIA mRNA has been reported to be downregulated in both females and males. It is concluded that the regulation of MyHC isoforms by thyroid hormone differs between females and males, presumably as a result of a gender-related difference in the translational or post-translational regulation of MyHC synthesis.

Oxidative Enzyme Activity and Soma Size in Motoneurons Innervating the Rat Slow-Twitch and Fast-Twitch Muscles After Chronic Activity

The effects of chronic activity induced by running training on the activity of the mitochondrial enzyme succinate dehydrogenase (SDH) and soma size in motoneurons innervating the slow-twitch soleus (SOL) and fast-twitch extensor digitorum longus (EDL) muscles were studied in rats using the retrograde neuronal tracer Nuclear Yellow. Rats were assigned to control and trained groups that were subjected to treadmill running for 10 weeks (2 h/day, 30 m/min, 5 days/week). After training, both SOL and EDL muscles showed clear adaptations (citrate synthase activity in the SOL muscle, and the fast-twitch oxidative-glycolytic fiber area of the EDL muscle increased significantly after training). The SDH activity of the motoneurons innervating both SOL and EDL muscles was unchanged by training. However, SOL motoneurons of trained rats had a significantly larger soma size and a significantly higher total SDH activity (SDH activity × soma size) than those of control. Total SDH activity was calculated to examine the absolute SDH protein content of the motoneurons. On the other hand, there was no difference in both soma size and total SDH activity of EDL motoneurons between the two groups. These data demonstrate that chronic activity has a considerably stronger impact on soma size and total oxidative enzyme activity of motoneurons innervating slow-twitch rather than fast-twitch muscles.

Coordinated fast-to-slow transitions of myosin and SERCA isoforms in chronically stimulated muscles of euthyroid and hyperthyroid rabbits.

Changes in the patterns of myosin heavy chain (MHC) isoforms, isomyosins, and Ca(2+)-ATPase (SERCA) isoforms were studied in long-term (72 d) stimulated fast-twitch extensor digitorum longus (EDL) and tibialis anterior (TA) muscles of euthyroid and hyperthyroid rabbits. The chronic low-frequency stimulation-induced fast-to-slow transitions in MHC isoforms, isomyosins and SERCA isoforms were pronounced in muscles from euthyroid rabbits, but less pronounced in muscles from hyperthyroid rabbits. Thus, hyperthyroidism counteracted to same extent the stimulation-induced fast-to-slow transition. Analyses of all parameters were performed on the same individual muscles, providing information on the co-ordinated expression of SERCA and myosin isoforms. A high correlation (r = 0.97) was detected between relative concentrations of slow SERCA2a and slow MHCI isoforms. This correlation persisted under all experimental conditions, suggesting a co-ordinated expression of slow myosin and Ca(2+)-ATPase isoforms. Conversely, fast SERCA1a was correlated to fast myosin isoforms as a whole.