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The relation between ATPASE activity and light chains of myosin in developing, adult and denervated muscles of several animals species.

Ca2+ATPase activity and light chains of myosin, fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, in developing, adult and denervated fast, slow and cardiac muscles of the rat, guinea-pig, cat, rabbit and chick were studied. It has been shown that in normal adult muscles the electrophoretic pattern of light chains of myosin reflects the myosin ATPase activity only when muscles from the same animal species are compared. In homologous muscles from adult animals differing in size, the size-dependent difference in myosin ATPase activity is not revealed in the electrophoretic pattern. Both in developing and in denervated muscle, changes in myosin ATPase activity are either connected with changes in the pattern of light chains of myosin or this pattern does not change. This relation is different in fast and slow muscles and also differs in chick and rabbit muscles. There are several possibilities of explaining the relation between ATPase activity of myosin and the pattern of light chains of myosin. The observation that myosin from the soleus muscle of 1-month-old rabbit contains light chains corresponding to both fast and slow type of myosin, indicates that the change in myosin ATPase activity during development is due to changes in the ratio between the fast and slow type of myosin.

Adenosine Triphosphatases↗

The effects of amantadine, rimantadine and levodopa on the frequency of fibrillation potentials in denervated muscle.

Right mid-thigh sciatic denervation was performed in 59 rats. Eleven days later they were assigned into 4 groups: a control group that was injected intraperitoneally with normal saline, and 3 test groups that were injected intraperitoneally with amantadine, rimantadine or levodopa (50 mg/kg each), for 3 consecutive days. On the fourteenth day, 2 h after the third injection, electromyography of the soleus-gastrocnemius muscles was performed under pentothal anesthesia. Fibrillation potentials were counted. The statistical analysis showed significant reduction of fibrillations secondary to levodopa and to amantadine injections. Rimantadine did not change the frequency of fibrillations. A stabilizing effect of levodopa and amantadine on the denervated plasmalemma is postulated.

Acetylcholine↗

Denervated muscle: rate of propagation and effects of manganese ions and glycerol on the fibrillatory activity of frog semitendinosus.

Denervated frog's semitendinosus muscles were studied. Fibrillation potentials generate propagated spikes which are conducted at a rate similar to that calculated for innervated muscles. The twitch tension per unit area was similar in denervated and innervated single fibres; the average value corresponded with values reported in the literature for fibres stimulated at 125/sec. Manganese ions (1-3 mM), selectively inhibit fibrillation potentials and have no appreciable effect upon evoked spikes. Mn2+ also inhibits (40-50%) the twitch responses and abolishes the resting tension of muscle fibres. The latter effects were observed in denervated as well as in innervated preparations when the fibres studied belonged to fasicles located at the site of entrance of the nerve. Disruption of the transverse tubular system by glycerol treatment does not interfere with the generation of fibrillation potentials, although the effects on twitching are similar to those reported in the literature. A possible mechanism is proposed to explain the generation of fibrillation potentials in frog denervated muscles.

Animals↗

Improved functional recovery of denervated skeletal muscle after temporary sensory nerve innervation.

Prolonged muscle denervation results in poor functional recovery after nerve repair. The possible protective effect of temporary sensory innervation of denervated muscle, prior to motor nerve repair, has been examined in the rat. Soleus and gastrocnemius muscles were denervated by cutting the tibial nerve, and the peroneal nerve was then sutured to the transected distal tibial nerve stump either immediately or after two, four or six months. In half of the animals with delayed repair, the saphenous (sensory) nerve was temporarily attached to the distal nerve stump. Muscles were evaluated three months after the peroneal-to-tibial union, and were compared with each other, with unoperated control muscles and with untreated denervated muscles. After four to six months of sensory "protection", gastrocnemius muscles weighed significantly more than unprotected muscles, and both gastrocnemius and soleus muscles exhibited better preservation of their structure, with less fiber atrophy and connective tissue hyperplasia. The maximum compound action potentials were significantly larger in gastrocnemius and soleus muscles following sensory protection, irrespective of the delay in motor nerve union. Isometric force, although less than in control animals and in those with immediate nerve repair, remained reasonably constant after sensory protection, while in unprotected muscles there was a progressive and significant decline as the period of denervation lengthened. We interpret these results as showing that, although incapable of forming excitable neuromuscular junctions, sensory nerves can nevertheless exert powerful trophic effects on denervated muscle fibers. We propose that these findings indicate a useful strategy for improving the outcome of peripheral nerve surgery.

Animals↗

[The effects of clenbuterol on intramuscular collagen metabolism in denervated muscle].

In order to explore the effects of clenbuterol on intramuscular collagen metabolism in denervated skeletal muscles, a randomized, double-masked and placebo-controlled group were studied. Seventy-one patients with complete function loss in muscularcutaneous nerve resulted from brachial plexus injury were administered clenbuterol or placebo 60 micrograms Bid for more than 3 months. Biopsies of the biceps brachia muscle were performed at the beginning and end of this study. The biopsied muscles were processed with anti-collagen I and IV immunohistochemical stains and image analysis as well. The result showed that the collagen proliferation of both type I and IV was much reducible in the clenbuterol-treated group than that of the placebo-treated group (P < 0.05). It was concluded that clenbuterol could inhibit partially the proliferation of intramuscular collagens in denervated skeletal muscle.

Adolescent↗

A stimulator for functional activation of denervated muscles.

In recent years various studies proved that electrical stimulation can improve contractile capability and restore muscle function in long-term denervated degenerated muscles. The low excitability of the muscle cells at the initial stage of training and surrounding connective tissue, acting as an electrical shunt, require special stimulation parameters. Until now, no appropriate devices (stimulators) are commercially available. Therefore, we were forced to design our own stimulators. The control unit of the stimulators is based on a microprocessor for maximum flexibility regarding the generation of the parameters such as pulse amplitudes, pulse width, frequency, stimulation times, ramps, and so on. In addition, the microprocessor design allows recording of compliance data such as stimulation date, time, duration, and used programs. The constant voltage output stage of the stimulator is able to generate biphasic charge balanced stimulation impulses with a pulse width of 1 to 300 ms, voltage amplitudes up to +/-80 V (160 VPP), and stimulation currents up to 250 mA. To prevent direct current due to inexact charge compensation, the electrode outputs are decoupled capacitively. Simultaneous 2 channel stimulation with independent intensity levels is possible. The stimulators are programmed using a notebook or a personal digital assistant via infrared serial interface. This concept guarantees the application of correct stimulation parameters because the patient has only access to parameters that are preprogrammed for him in the outpatient clinic. For the home based training, access is limited to variation of intensity within preprogrammed limits. For safety reasons, the portable unit is powered by an internal rechargeable battery. High efficiency switched voltage regulators are used to provide the different required voltage levels while ensuring an acceptable operating time of the stimulator.

Electric Power Supplies↗

Disturbed trafficking of dystrophin and associated proteins in targetoid phenomena after chronic muscle denervation.

Dystrophin and associated proteins form a complex with an important role at the sarcolemma. Expression of this protein complex is highly regulated during development and regeneration. In order to better understand assembling patterns of these proteins, we have studied their expression in targetoid-like phenomena found in human muscle after chronic denervation, a situation known to give rise to abnormal protein trafficking. In eight biopsies of patients with chronic denervation, mainly resulting from amyotrophic lateral sclerosis, we found a number of targetoid phenomena. Selective accumulation of a number of sarcolemmal and sarcoplasmatic proteins occurred in targetoid phenomena. The larger majority of them contained gamma-sarcoglycan (gammaSG), but none contained the developmental heavy chain myosin isoform. In a series of 166 targetoid phenomena which could be studied with 17 different antibodies recognizing sarcolemmal and cytoplasmatic proteins, a high level of colocalization of gammaSG with desmin and alpha-actinin was found. Colocalization rate was much lower with other proteins, including other members of the dystrophin-associated protein complex. These data show that selective changes in expression of otherwise closely related proteins occur during disturbed trafficking leading to target formation. Because members of the dystrophin-associated protein complex do not accumulate in a similar fashion within targets, we suggest that a complex molecular control of gene expression and trafficking of this complex is involved after chronic muscle denervation.

Amyotrophic Lateral Sclerosis↗

Localization of neural cell adhesion molecule in denervated muscle to both the plasma membrane and extracellular compartments by immuno-electron microscopy.

The location of neural cell adhesion molecule in mouse skeletal muscle has been investigated using two immuno-electron microscopical techniques. In the first method, 6-micron frozen sections of a normal and a hemi-denervated gluteus muscle were stained by an indirect immunoperoxidase method using a rabbit-derived polyclonal antiserum to neural adhesion molecule as the primary antibody. The stain was visualized with the electron-dense chromogen, diaminobenzidine and the sections fixed, osmicated and processed for electron microscopy whilst still on the slide. Ultrathin sections were cut and viewed in the electron microscope, where the reaction product appeared to be localized in the plasma membrane and on the basal lamina of the muscle fibres. The second method, using a 5-nm immunogold-labelled secondary antibody, confirmed and extended these initial observations. In these experiments, the primary antibody to neural cell adhesion molecule was applied in vivo to hemi-denervated mouse gluteus muscles. The muscles were dissected out 24 h later, divided into denervated and innervated halves and then into junctional and extrajunctional regions. Together with the junctional and extrajunctional regions of normal, control pieces of muscle which had not been incubated with anti-neural cell adhesion molecule, they were lightly fixed, and incubated with the secondary gold-labelled antibody, before further fixation and processing for electron microscopy. Semi-thin, 0.5-micron sections of each were cut and viewed at 20,000 x magnification. Randomly sampled fibres from the extrajunctional regions of the denervated and innervated halves of the hemi-denervated gluteus and the normal, control gluteus were photographed for quantitative analysis. Five micrographs were randomly selected from each group and in these, the numbers, density and position of gold particles were measured using a digitizing tablet. By far the highest number of gold particles was found in the denervated half of the gluteus muscle, there being much less in the innervated half and practically none in the control. The density of gold particles in the denervated muscles was maximal in the plasma membrane-basal lamina complex, but most were located in the extracellular spaces outside this region, where they usually occurred in clusters apparently in association with collagen fibres. We conclude that neural cell adhesion molecule in denervated skeletal muscle is present not only in the plasma membrane but also in the basal lamina and extracellular space.

Animals↗

Direct neurotization of severely damaged denervated muscles.

Previously, when part of a muscle and the terminal part of its nerve with its branches were removed through trauma, no technique for repair was available. A new technique for direct neurotization of denervated muscles is presented. This consists of the insertion of the distal stump of a nerve graft, divided into many small branches, directly into the muscle. Very good results were obtained in eight clinical cases of severely damaged muscles which were operated on according to this technique.

Humans↗

Effect of estrogen on denervated muscle.

The rate of increase of glucose 6-phosphate dehydrogenase activity in denervated rat extensor digitorum longus muscle shows sexual dimorphism. This phenomenon is further investigated in this report by assessing the effects of ovariectomy, hypophysectomy, hormone replacement therapy, and treatment with an estrogen antagonist , MER-25. The data demonstrate that physiologic doses of estrogens enhance the rate and extent of the increase in glucose 6-phosphate dehydrogenase activity after denervation. The data further indicate that aromatization of androgens may be a significant source of estrogen involved in hormonal modulation of the neural control of glucose 6-phosphate dehydrogenase and other processes in muscle. Furthermore, choline acetyltransferase activity, a marker for the neuromuscular synapse, decreased in rat extensor digitorum longus muscles after denervation, but was unaffected by ovariectomy.

Animals↗

Value of magnetic resonance imaging for muscle denervation syndromes of the shoulder girdle.

Clinical evaluation of neuromuscular disorders typically consists of obtaining a detailed clinical history, physical examination, and electrophysiologic examinations. Electrodiagnostic examinations significantly aid in distinguishing between myopathy, neuropathy, and neuromuscular disorders. Electrodiagnostic examinations also assist in determining the severity and extent of disease. Progress can also be monitored on follow-up testing. The benefit of magnetic resonance imaging (MRI) in neuromuscular disease evaluation lies primarily in identifying a specific underlying gross pathologic cause and its location in the neuraxis as well as in identifying associated secondary findings. In some cases, MRI is particularly helpful when a solitary, small, deep muscle is affected. Imaging can be useful in assessing clinical progress in some cases. Causes of muscle denervation include mass lesions and trauma as well as infectious, autoimmune, and idiopathic causes. This article illustrates the common denervation syndromes that involve the shoulder girdle: Parsonage-Turner syndrome, quadrilateral space syndrome, and suprascapular neuropathy. By demonstrating the exact muscles involved and spared, MRI noninvasively identifies the level of nerve insult in the neuraxis. Furthermore, in cases in which a mass is responsible for denervation, MRI can directly show the cause and aid in treatment planning.

Humans↗

In vivo assessment of conduction velocity and refractory period of denervated muscle fibers.

Stimulation needle electromyography was used to study the muscle fiber conduction velocity and refractory period in 4 patients with long-term denervation of the lower limb muscles due to lesion of the conus cauda or cauda equina (2 untrained and 2 trained by functional electrical stimulation). In untrained patients, the results demonstrated that propagation velocity is reduced and refractory period of the muscle fiber is increased with time of denervation. The patients performing electrical stimulation training showed higher conduction velocities and reduced refractory period despite longer lasting denervation. This suggests that electrical stimulation training is effective to improve the electrical properties of the muscle fiber. Since the obtained data show a good correlation to other clinical tests and biopsy investigations, this method could serve as an additional measurement technique to specify the status of the denervated muscle. Further animal experiments and clinical studies are necessary to proof the results in comparison to more invasive established techniques.

Electric Stimulation↗