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PubMed1981

Ca2+-uptake properties of two populations of mitochondria from normal and denervated rat soleus muscle.

Ultraturrax- and Nagarse-released populations of mitochondria were characterized with respect to their Ca2+-uptake activities (i) by means of the indirect polarographic technique and (ii) directly by the 45Ca Ruthenium Red-quench method of Reed & Bygrave [(1974) Biochem. J. 140, 143-155]. The denervated-muscle subsarcolemmal and intermyofibrillar mitochondrial fractions displayed markedly decreased rates and capacities for Ca2+ uptake compared with their respective controls. The implications of these findings with respect to the process of cell necrosis are discussed.

Animals↗
PubMedDate not supplied

Fasciculation frequency changes at different length of relaxed muscle.

We have studied, in chronically denervated muscles, fasciculation potentials (FPs) which were clearly identified when the muscles were either relaxed or passively stretched. The frequency of 10 FPs was determined in both positions in seven patients. In stretched muscles, two FPs were no longer recorded, and the mean FP frequency decreased significantly (P < .01). There was also a decrease of the FP amplitude. As FPs are frequently associated with ordinary muscle cramps, stretching of the muscle may lessen the frequency of FPs by the same mechanism which stops cramps.

Adult↗
PubMed1978

Effects of alpha-bungarotoxin and reversible cholinergic ligands on normal and denervated mammalian skeletal muscle.

alpha-Bungarotoxin (BuTX; 5 micrograms/ml) completely blocked the endplate potential and extrajunctional acetylcholine (ACh) sensitivity of surface fibers in normal and chronically denervated mammalian muscles, respectively, in about 35 min. A 0.72 +/- 0.033 mV amplitude endplate potential returned in normal muscle fibers after 6.5 hr. of washout of alpha-BuTX, and an ACh sensitivity of 41.02 +/- 3.95 mV/nC was recorded in denervated muscle after 6.5 hr of wash (control being 1215 +/- 197 mV/nC). A two-step reaction of BuTX with binding sites which may allosterically interact is postulated. Several pharmacologic differences were noted between the ACh receptors at the normal endplate and those appearing extrajunctionally following denervation. In normal innervated muscles exposed to BuTX in the presence of 20 microM carbamylcholine or decamethonium, washout of both drugs restored twitch to control levels within 2 hr. Endplate potentials large enough to initiate action potentials were also recorded in most surface fibers. In contrast, these agents, in much higher concentrations (50 microM), were almost ineffective in preventing BuTX blockade of ACh sensitivity in denervated muscle. Hexamethonium (10 and 50 mM) depressed neuromuscular transmission and blocked the action of BuTX in normal muscle in a dose-dependent fashion. On the extrajunctional receptors, hexamethonium (50 mM) was ineffective in protecting against BuTX. We may conclude that at the normal endplate region there are two distinct populations of ACh receptors, both of which react with cholinergic ligands and BuTX, but that a small population (representing congruent to 1% of the total) reacts with BuTX reversibly. Our findings further suggest a clear distinction between ACh receptors located at the normal endplate region and those of the extrajunctional region of the chronically denervated mammalian muscle.

Acetylcholine↗
PubMed2005

Asymmetric atrophy of the supraspinatus muscle following tendon tear.

Muscle atrophy is a known consequence of muscle disuse, muscle denervation and tendon tear. Whereas after nerve injury muscle atrophies in the denervated area, the distribution of muscle atrophy following tear of its tendon is not known. Standardized MRI scans of 64 consecutive, painful shoulders were evaluated for supraspinatus tendon tearing, myotendinous retraction, supraspinatus muscle atrophy, fatty infiltration, ratio of the scapular (deep) and fascial (superficial) muscle area ("symmetry") and position of the central tendon within the supraspinatus fossa. There were thirteen shoulders with no and eleven shoulders with partial thickness supraspinatus tendon tears. In the forty cases with full thickness tendon tear, there was significant muscle atrophy and fatty infiltration. Atrophy of the fascial muscle portion was 43%, on the bony side it was 9% (p<0.005). The position of the central tendon within the supraspinatus fossa, was unaltered. Muscular changes following tendon tear occur highly asymmetrically: the muscle portion originating from the fascia primarily atrophies, the portion originating from the scapula primarily undergoes fatty infiltration. Muscular changes are not simply a consequence of muscle disuse, but dependent on architectural changes in the muscle.

Adipose Tissue↗
PubMed1993

Differential expression of muscle regulatory factor genes in normal and denervated adult rat hindlimb muscles.

Skeletal muscle represents an excellent model system in which to examine regulatory mechanisms that modulate gene expression in the mature adult organism. Individual muscle fibers can be categorized as fast- or slow-twitch based upon several physiological and molecular criteria, including metabolic enzyme activity and contractile protein isoforms. Each property can be influenced by a variety of factors such as changes in motor neuron activity or alterations in hormone levels, although the molecular pathways by which environmental factors affect gene expression remain largely unknown. As a first step in identifying potential regulators of fiber-type diversity, the expression patterns of four basic/helix-loop-helix muscle regulatory factors (MRFs), referred to as MyoD, myogenin, Myf-5, and MRF4, were examined in normal adult rat muscles which differed in their phenotypic properties. As expected, all four MRFs were expressed at detectable levels in the muscles studied. However, different muscles accumulated different proportions and combinations of MRF transcripts. For example, myogenin expression was maximally detected in slow-twitch muscles whereas MyoD transcripts were found predominantly in muscles exhibiting a fast-twitch phenotype. Induced phenotypic changes in two fast-twitch muscles via denervation lead to a large and rapid increase in transcript levels of all four MRFs as early as 24 hr following denervation, with myogenin transcripts approaching 150-200-fold higher levels than innervated contralateral muscles within 7 days. These results suggest that myogenin, as well as the other three MRFs, may be involved in both the initial establishment as well as maintenance of fiber-type diversity in the developing organism.

Animals↗
PubMed2005

Effect of salbutamol on innervated and denervated rat soleus muscle.

The objective of the present investigation was to perform a 14-day time-course study of treatment with salbutamol, a beta2 adrenoceptor agonist, on rat soleus muscle in order to assess fiber type selectivity in the hypertrophic response and fiber type composition. Male Wistar rats were divided into four groups: control (N = 10), treated with salbutamol (N = 30), denervated (N = 30), and treated with salbutamol after denervation (N = 30). Salbutamol was injected intraperitoneally in the rats of the 2nd and 4th groups at a concentration of 0.3 mg/kg twice a day for 2 weeks. The muscles were denervated using the crush method with pean. The animals were sacrificed 3, 6, 9, 12, and 14 days after treatment. Frozen cross-sections of soleus muscle were stained for myosin ATPase, pH 9.4. Cross-sectional area and percent of muscle fibers were analyzed morphometrically by computerized image analysis. Treatment with salbutamol induced hypertrophy of all fiber types and a higher percentage of type II fibers (21%) in the healthy rat soleus muscle. Denervation caused marked atrophy of all fibers and conversion from type I to type II muscle fibers. Denervated muscles treated with salbutamol showed a significantly larger cross-sectional area of type I muscle fibers, 28.2% compared to the denervated untreated muscle. Moreover, the number of type I fibers was increased. These results indicate that administration of salbutamol is able to induce changes in cross-sectional area and fiber type distribution in the early phase of treatment. Since denervation-induced atrophy and conversion from type I to type II fibers were improved by salbutamol treatment we propose that salbutamol, like other beta2 adrenoceptor agonists, may have a therapeutic potential in improving the condition of skeletal muscle after denervation.

Adrenergic beta-Agonists↗
PubMed1993

Denervated human skeletal muscle: MR imaging evaluation.

Because determination of neurologic integrity after severe limb trauma is crucial in patient care, the authors assessed magnetic resonance (MR) imaging as a tool to map denervated motor units of skeletal muscle in patients with traumatic peripheral neuropathy. Denervation was confirmed in 22 patients with use of electromyography, surgery, or both. MR imaging was performed with moderately T1- and T2-weighted spin-echo and short-tau inversion-recovery (STIR) sequences. MR imaging was unreliable in depicting acute denervation. Muscles of patients with subacute denervation had prolonged T1 and T2, which contributed to conspicuous hyperintensity on STIR images. Chronically denervated muscles showed marked atrophy, variable changes on STIR images, and conspicuous fatty infiltration on T1-weighted images. Normal variants in motor unit anatomy were seen in denervated muscle volumes outside the expected distribution of the injured nerve. MR imaging is promising for the noninvasive mapping and monitoring of denervated muscle in subacute and chronic phases of peripheral neuropathy.

Extremities↗
PubMed1977

Partial denervation affects both denervated and innervated fibers in the mammalian skeletal muscle.

Partial denervation of the rat extensor digitorum longus muscle was performed by sectioning only one of the sciatic nerve roots. Measurements of spike resistance to tetrodotoxin in individual muscle fibers revealed denervation changes not only in the denervated fibers but also in the adjacent innervated ones. The results support the concept that products of nerve degeneration play a role in the origin of muscle changes induced by denervation.

Action Potentials↗
PubMed1987

Effect of malathion on some kinesiological parameters of the gastrocnemius muscle of frog, Rana hexadactyla (Lesson) and the antagonistic action of atropine.

Selected kinesiological parameters were studied in the gastrocnemius muscle of frog, R. hexadactyla Lesson, under different treatments with malathion and atropine. There is an apparent decrease of twitch amplitude, half contraction time, half relaxation time, twitch duration and fatigue time in control muscle during malathion treatment. In the denervated muscle, twitch amplitude, latency period, and fatigue time were lowered, with elevated phases of contraction and relaxation time. However, all these parameters except fatigue time showed an elevatory trend when atropine was given along with malathion revealing its antagonistic action against malathion toxicity. All these results indicate that the action of malathion is similar to that of a pharmacologically active myo- and neurotoxic agent.

Animals↗
PubMed1983

The regulation of extrajunctional acetylcholine receptors in the denervated rat diaphragm muscle in culture.

The regulation of the number of extrajunctional acetylcholine (ACh) receptors was assayed by 125I-labelled alpha-bungarotoxin binding sites in denervated rat diaphragm muscle in culture. Sustained K depolarization does not eliminate extrajunctional ACh receptors. In fact, muscle cultured in high-K medium (normal Cl) for 3 days exhibits a greater binding capacity than controls. Under conditions in which the intracellular Cl concentration is unaltered (high-K-low-Cl medium) this effect of high-K medium on the number of extra-junctional ACh receptors is blocked. The number of extrajunctional receptors increases 24-48 h after exposure to high-K-normal Cl medium, similar to the time course of the initial appearance of extrajunctional receptors in the denervated diaphragm muscle in vivo or in organ culture in normal media. High-K-normal Cl medium did not alter the rate of receptor degradation. Electrical stimulation of denervated muscle strips cultured in low-Ca medium containing D-600 eliminated extrajunctional receptors as efficiently as stimulation of muscles in control medium. Electrical stimulation did not reduce the extrajunctional ACh receptor population in glycerol-treated uncoupled muscles to the same extent as in untreated muscles. The extrajunctional ACh receptor content of denervated muscle cultured for 3 days in 2 and 5 mM-caffeine was reduced by about half respectively. Denervated muscle cultured in 0.3 mM-caffeine did not differ from control denervated muscle. Other agents which may alter intracellular cyclic nucleotide levels: dibutyryl cyclic GMP, dibutyryl cyclic AMP, papaverine, and sodium nitroprusside, did not mimic the effect of caffeine or electrical stimulation in lowering the levels of extrajunctional ACh receptors. We conclude that intracellular Ca release from the sarcoplasmic reticulum is necessary for the elimination of extrajunctional ACh receptors in denervated muscle. The levels of intracellular Cl also influence the population of extrajunctional receptors. Conditions which lead to higher levels of intracellular Cl result in greater rates of synthesis of ACh receptors.

Action Potentials↗
PubMed2001

AMP-activated protein kinase activation prevents denervation-induced decline in gastrocnemius GLUT-4.

This study was designed to determine whether the reductions in GLUT-4 seen in 3-day-denervated muscles can be prevented through chemical activation of 5'-AMP-activated protein kinase (AMPK). Muscle AMPK can be chemically activated in rats using subcutaneous injections with 5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside (AICAR). In this study, the tibial nerve was sectioned on one side; the other was sham operated but without nerve section. Acute injections of AICAR resulted in significantly increased AMPK activity in denervated gastrocnemius but not soleus muscles. Acetyl-CoA carboxylase activity, a reporter of AMPK activation, declined in both gastrocnemius and soleus in both denervated and contralateral muscles. Three days after denervation, GLUT-4 levels were significantly decreased by approximately 40% in gastrocnemius muscles and by approximately 30% in soleus muscles. When rats were injected with AICAR (1 mg/g body wt) for 3 days, the decline in GLUT-4 levels was prevented in denervated gastrocnemius muscles but not in denervated soleus muscles. The extent of denervation-induced muscle atrophy was similar in AICAR-treated vs. saline-treated rats. These studies provide evidence that some effects of denervation may be prevented by chemical activation of the appropriate signaling pathways.

AMP-Activated Protein Kinases↗
PubMed1975

Further studies on the binding properties of cobra neurotoxin to cholinergic receptors in mouse skeletal muscle.

Preparations of 3H-monoacetylated derivatives of Naja naja siamensis neurotoxin siamensis 3 were found to be resistant to degradation and deacetylation during in vitro muscle incubation. A low rate of free 3H-acetate (less than 0.4%) was present in the preparations, but should not interfere with the binding of toxin to cholinergic receptors in mouse extensor digitorum longus muscles in vitro. The binding of toxin to cholinergic receptors in mouse extensor digitorum longus muscle was essentially irreversible. d-Tubocurarine antagonized the binding of toxin in both innervated and denervated muscles. Administration of actinomycin D one day after denervation prevented the appearance of new toxin binding sites in the muscles.

Animals↗
PubMed1975

Histochemical and neurophysiological studies of autotransplanted cat muscle.

Free autologous transplants of previously denervated whole muscles in adult cats were studied with histochemical and electromyographical methods. The transplants were placed in contact with normal intercostal muscles in intercostal spaces and removed at intervals from 5 days to 44 weeks after transplantation. Normal muscles as well as transplanted muscles without previous denervation and muscles denervated and left in situ were also studied.. The fibres of the previously denervated grafts survived the first avascular period, but a de-differentiation of enzymatic characteristics of the red and white fibres occurred and endured during the first 6 weeks after grafting. The first signs of reinnervation could be identified with electromyography after 4 weeks and with histochemical techniques after 6. Reinnervation of most of the fibres of the transplants was completed about 14-18 weeks after grafting. No fibrillations were then found electromyographically and in the histochemical preparations most fibres had attained near-normal staining characteristics. Type grouping was apparent but less pronounced than reported after reinnervation through the motor nerve. There was conclusive evidence that previously denervated muscles survived transplantation and became structurally and enzymatically mature whereas muscles transplanted without previous denervation undergo extensive fibrotic changes.

Adenosine Triphosphatases↗
PubMed1988

Effect of denervation and nerve extract on ultrastructure of muscle.

Changes in denervated skeletal muscle result both from disuse and loss of neurogenic trophic substances. It had been shown that administration of nerve extract intramuscularly in rats or systemically in mice prevented the nondisuse component of atrophy in denervated muscle. Amelioration of atrophy was manifested as reduced losses of weight, protein, and cross-sectional areas of fiber in denervated hind-limb muscles. The present study assessed the effects of nerve extract on ultrastructural changes in different types of fiber (as classified by activity of ATPase) in mouse skeletal muscle denervated for 7 days. Denervated and contralateral innervated muscles of treated and untreated mice were examined by electron microscopy, and morphological parameters were quantitated by stereological techniques. Denervated muscles exhibited smaller reductions of several ultrastructural changes in treated than in untreated mice including sizes of mitochondria, and percentage volume per fiber of mitochondria, sarcoplasmic reticulum, and t-tubules. The magnitude of the myotrophic effects varied in the different types of fiber, with amelioration of between 50 and 95% of the postdenervation changes.

Animals↗
PubMed1985

Two subtypes of sodium channel with tetrodotoxin sensitivity and insensitivity detected in denervated mammalian skeletal muscle.

The action potential (AP) in most nerve and muscle preparations depends upon nanomolar concentrations of the neurotoxins saxitoxin (STX) and tetrodotoxin (TTX). In some excitable tissues lacking mature innervation, a toxin-resistant AP has been described by electrophysiological results. However, multiple attempts to detect corresponding toxin-resistant Na channels with radiolabelled STX and TTX have been unsuccessful. We report here the detection of Na channels with low-affinity binding of STX and TTX, accounting for 50-60% of the Na channels in rat hindlimb muscle 4-5 days after denervation.

Action Potentials↗
PubMed1986

Effects of short-term denervation on avian muscle spindle structure.

The role of the nerve in maintaining the ultrastructural integrity of avian muscle spindles was investigated by denervating the pigeon's extensor digitorum communis for periods of 10, 19, and 28 days. The equatorial region of control intrafusal fibers had a reduced density of myofilaments. Sensory endings contained mitochondria and structures resembling synaptic vesicles, and were associated with satellite cells. In the polar region, fibers had a high concentration of myofilaments; small motor endings, unlike sensory endings, lay outside of the fiber's basal lamina. The outer capsule consisted of thin, tightly layered cells which gradually became reduced in number distal to the equatorial region. In both equatorial and polar regions the capsule became more disrupted with longer denervation periods, and lysosomes and phagocytes became more abundant. The equatorial region of denervated fibers contained many myofibrils and some had peripherally-located nuclei, unlike the controls; sensory terminals were absent. The polar region of some fibers had disorganized myofilaments and others had a reduced myofilament density. Fiber diameters increased significantly in both regions. Thus, denervated intrafusal fibers lost some characteristics which distinguish them from extrafusal fibers.

Animals↗
PubMed1992

Effect of clenbuterol on protease activities and protein levels in rat muscle.

To elucidate the biochemical mechanism by which the sympathomimetic agent clenbuterol promotes skeletal muscle growth, we have determined the activity of a range of proteolytic enzyme types (acid, neutral and alkaline proteinases and peptidases), together with the levels of soluble and structural proteins (via SDS-polyacrylamide gel electrophoresis) in 5 innervated and denervated muscle types from control and drug-treated rats. No gross change in activity was found for any enzyme type in any muscle (innervated or denervated) following clenbuterol treatment; however, one enzyme, arginyl aminopeptidase, showed a small but consistent decrease in activity in all of the innervated muscles investigated. Similar fractionation profiles were obtained for structural or soluble proteins from corresponding muscle types (innervated or denervated) in control or clenbuterol-treated animals, with the exception of cardiac muscle, which showed a 50% increase in staining intensity of one band (subunit molecular mass 18 kD). We conclude that the anabolic action of clenbuterol in promoting skeletal muscle growth does not occur via downregulation of protease activity, or increase in levels of individual muscle proteins.

Animals↗