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PubMed1984

[Effect of hydrocortisone on the membrane properties of innervated and denervated muscle fibers in frogs].

Two injections of 10, 100 or 1000 micrograms of hydrocortisone into the lymphatic sack with one-week interval or daily injections of 100 micrograms of the hormone did not hinder a decrease in the membrane resting potential, an increase of the input resistance and time constant of the membrane as well as appearance of nonsynaptic sensitivity to acetylcholine in muscle fibers developing by the 13-15th day after denervation of the muscle. In latter two cases, however, no increase of the input resistance and time constant of the membrane occurs. Administration of the same doses of drug to intact frogs led to a decrease of the membrane resting potential, increase of the time constant and input resistance of the membrane whereas daily injections reduced the sensitivity of muscle fibers to acetylcholine. Hydrocortisone seems to be unable to hinder the postdenervation changes in the muscle membrane whereas high doses of the hormone are able to induce changes in the muscle membrane.

Animals↗
PubMed2002

Effects of nerve growth factor on the neurotization of denervated muscles.

Studies on surgical repair techniques of the peripheral nerve are still trying to improve the outcome. There are many studies on the effects of various neurotrophic factors on the transected peripheral nerve. Muscular neurotization, which is the direct implantation of the nerve to the target denervated skeletal muscle, is one of the techniques used when the primary repair of the peripheral nerves is not possible. The effects of nerve growth factor (NGF), which is one of the primary neurotrophic factors, on the reinnervation of denervated muscles by neurotization is investigated in this experimental study. The denervated soleus muscle was neurotized via peroneal nerve implantation (group 1), and NGF was administered to the neurotized muscle (group 2). All animals were evaluated at weeks 8, 10, and 12 using electromyography. Muscle contractility, muscle weight, and histological morphometric tests were performed at week 12. The experimental groups were compared with each other and normal control values. Electromyographically, group 2 (direct nerve implantation + NGF) demonstrated better reinnervation in all evaluations. The study of muscle weight showed that the muscle mass was 75% of the normal soleus muscle in group 1 and was 85% of the normal side in group 2 at the end of week 12. In group 1, the twitch force was 56% of the normal soleus muscle and was 71% in group 2. Tetanic force was 53% of the normal soleus muscle in group 1 and 68% in group 2. Histological morphometric studies revealed that there was a decrease in the density of the motor end plates in group 1, but there was no statistically significant difference between the normal soleus muscles and the NGF applied to group 2. The positive effects of NGF on the neurotization of denervated muscles seen in this study suggest that it may be useful for treating some difficult reconstructions caused by denervation.

Animals↗
PubMed1998

Matrix metalloproteinases MMP-2 and MMP-9 in denervated muscle and injured nerve.

Nerve crush or axotomy results in a transient or longterm denervation accompanied by remodelling in nerve, muscle and neuromuscular junctions. These changes include an increased turnover of several extracellular matrix molecules and proliferation of Schwann cells in injured nerves. Given the role of matrix degrading metalloproteinases MMP-2 and MMP-9 (gelatinases-type IV collagenases) in extracellular matrix remodelling, we investigated their regulation and activation in denervated muscles and injured nerves in mice. For this, immunofluorescence using MMP-2 and MMP-9 antibodies was carried concomitantly with gelatin zymography and quantification of gelatinase activity using [3H]-gelatin substrate. Results show that in normal mouse muscles MMP-2 and MMP-9 are localized at the neuromuscular junctions, in Schwann cells and the perineurium of the intramuscular nerves. In denervated mouse muscles, MMP-2 immunolabelling persists at the neuromuscular junctions but decreases in the nerves whereas MMP-9 immunolabelling persists at the neuromuscular junctions but is enhanced in degenerated intramuscular nerves. Denervated muscles did not show any significant change of gelatinolytic activity or expression pattern, while injured nerves exhibited a transient increase of MMP-9 and activation of MMP-2. In conclusion, this study demonstrates that MMP-2 and MMP-9 are expressed at mouse neuromuscular junctions and that their localization and expression pattern appear not to be modified by denervation. Their modulation in injured nerves suggests they are involved in axonal degeneration and regeneration.

Animals↗
PubMed1999

Relationship between hypoxemia and fibrillation potential firing rate in denervated muscle.

The effects of hypoxia and ischemia on the firing rate of fibrillation potentials in denervated rat muscle were examined. We recorded electromyograms from the denervated left extensor digitorum longus muscle. Hypoxia was induced by low-oxygen ventilation. Ischemia was established by ligating the abdominal aorta and inferior vena cava, with or without extracorporeal hindlimb perfusion. The fibrillation potential firing rate correlated with the PaO2 (P < 0.0001) and temperature (P = 0.0001). Fibrillation potentials disappeared after the initiation of ischemia and reappeared after restitution of blood flow; they disappeared during ischemia with extracorporeal perfusion. The attenuation curves for the firing rate of fibrillation potentials during ischemia were well-described by exponential curves, but there was no significant difference in the attenuation constants for circulatory arrest and perfusion with a physiologic salt solution. We conclude that the fibrillation potential firing rate is proportional to oxygen supply, presumably because of the rate of aerobic metabolism.

Action Potentials↗
PubMed1992

Alteration in the expression of GLUT-1 and GLUT-4 protein and messenger RNA levels in denervated rat muscles.

Denervation induces insulin resistance of the glucose transport process in skeletal muscle. To determine whether this is due to alterations in the expression of muscle glucose transporters (GLUT) in different fiber types, we evaluated the amount of GLUT-1 and GLUT-4 protein and messenger RNA (mRNA) in extensor digitorum longus (EDL) and soleus at 1, 2, and 3 days after sciatotomy. Denervation elevated the basal rate of 2-[1,2-3H]deoxy-D-glucose (2-DOG) uptake in the EDL and decreased the insulin-stimulated DOG uptake in both muscles. Denervation after 1 day did not modify the GLUT-1 or the GLUT-4 protein level in either muscle. However, it increased GLUT-1 mRNA by 66% and decreased GLUT-4 mRNA by 70% in the EDL, but not in the soleus (P < 0.05). After 2 days of denervation, by which time GLUT-1 mRNA was increased 2-fold and GLUT-4 mRNA was reduced by 70%, we observed a 2-fold increase in GLUT-1 protein (P < 0.01) in the EDL and a 40-45% decrease in GLUT-4 protein in both muscles (P < 0.01). These results indicate that modifications in the expression of GLUT-1 and GLUT-4 protein cannot explain the insulin resistance of the glucose transport process in the EDL or soleus 1 day after denervation. After 2 days of denervation, however, alterations in GLUT-1 and GLUT-4 protein levels may contribute to the change in basal and insulin-stimulated DOG uptake in both the EDL and the soleus muscles.

Animals↗
PubMed1981

Experimental rabies in skunks: persistence of virus in denervated muscle at the inoculation site.

Striped skunks (Mephitis mephitis) were inoculated into the denervated abductor digiti quinti muscle with street rabies virus. They were killed at various times after inoculation and several tissues were examined by immunofluorescence and light microscopy. Muscle at the inoculation site was examined electron microscopically. Rabies antigen was detected in muscle fibers first on day 7 and persisted until day 28. Light and electron microscopic lesions at the inoculation site included atrophic and degenerating muscle fibers and a few focal and regional endomysial accumulations of macrophages, lymphocytes and plasma cells. Scattered myocytes contained bodies of matrix, virions and anomalous tubular structures on electron microscopic examination. The results indicate that replication of rabies virus may occur in infected muscle fibers at the inoculation site until 28 days after exposure. This could contribute to variations in the incubation period for the first two to three months after exposure. However, the results do not support the contention that virus is contained in striated muscle cells throughout the long incubation periods.

Animals↗
PubMed1993

Persistent adrenergic reinnervation of previously denervated muscle in cat.

Synapse formation by aberrant nerves at chronically denervated intrinsic laryngeal muscles was observed even at 21 months after the transection of the recurrent laryngeal nerve in cats. Some of the aberrant nerves were adrenergic, and these nerve terminals were in synaptic contact with the postsynaptic membrane of the neuromuscular junctions together with non-adrenergic nerve terminals. Therefore, heterogeneous reinnervation between adrenergic nerve and muscle was found to be maintained for a long time.

Adrenergic Fibers↗
PubMed1991

Muscle denervation increases thyrotropin-releasing hormone (TRH) biosynthesis in the rat medullary raphe.

To determine whether thyrotropin-releasing hormone (TRH) could exert a trophic role in ventral horn motor neurons, we examined the effect of muscle denervation with botulinum toxin A on TRH mRNA in the rat medullary raphe by in situ hybridization histochemistry. Compared to controls, denervated rats showed a significant increase in the number and silver grain density of hybridized medullary raphe neurons. Increased proTRH gene expression in the medullary raphe in response to motor unit perturbation indicates that TRH may be trophic to lower motor neurons.

Action Potentials↗
PubMed2002

Denervated muscles in humans: limitations and problems of currently used functional electrical stimulation training protocols.

Prior clinical work showed that electrical stimulation therapy with exponential current is able to slow down atrophy and maintain the muscle during nonpermanent flaccid paralysis. However, exponential currents are not sufficient for long-term therapy of denervated degenerated muscles (DDMs). We initiated a European research project investigating the rehabilitation strategies in humans, but also studying the underlying basic scientific knowledge of muscle regeneration from satellite cells or myoblast activity in animal experiments. In our prior study, we were able to show that high-intensity stimulation of DDMs is possible. At the beginning of training, only single muscle twitches can be elicited by biphasic pulses with durations of 120-150 ms. Later, tetanic contraction of the muscle with special stimulation parameters (pulse duration of 30-50 ms, stimulation frequency of 16-25 Hz, pulse amplitudes of up to 250 mA) can improve the structural and metabolic state of the DDMs. Because there are no nerve endings for conduction of stimuli, large-size, anatomically shaped electrodes are used. This ensures an even contraction of the whole muscle. Contrary to the current clinical knowledge, we were able to stimulate and train denervated muscle 15-20 years after denervation. The estimated amount of muscle fibers that have to be restored is about 2-4 million fibers in each m. quadriceps. To rebuild such a large number of muscle fibers takes up to 3-4 years. Despite constant stimulation parameters and training protocols, there is a high variation in the developed contraction force and fatigue resistance of the muscle during the first years of functional electrical stimulation.

Electric Stimulation Therapy↗
PubMed1994

Electrophoretic separation of developmental and adult rabbit skeletal muscle myosin heavy chain isoforms: example of application to muscle denervation study.

We present the separation by SDS gel-electrophoresis of the six main myosin heavy chains (MHC) present in rabbit skeletal muscle. The separation of the four adult MHC (1, 2A, 2X/2D, 2B) was compared to that of the corresponding rat MHC as described by Talmadge and Roy [J. Appl. Physiol. 99 (1993) 2337-2340]. We found that many rabbit muscles contained mainly one of the four MHC, in some cases the 2B MHC. In addition, we resolved the embryonic E and perinatal P developmental MHC, which should facilitate muscle differentiation and regeneration studies in the rabbit. An example of application to the study of muscle denervation is given.

Animals↗
PubMed1980

The visualization of myosatellite cells in normal and denervated muscle: a new light microscopic staining technique.

A new light microscopic staining technique allows the visualization of satellite cells on the surface of myofibers. Either prior to or during fixation, whole frog sartorius muscles are bathed in an acidic buffered solution containing lead nitrate and subsequently exposed to ammonium sulfide. The staining of the satellite cells resulting from this procedure reveals their positions, and the outlines of their cell processes which occasionally branch. Electron microscopy shows that the staining is due to lead deposits localized between apposing membranes of satellite cells and associated myofibers. Prior exposure to N-ethyl-maleimide (NEM) does not alter the formation of the lead deposits on the satellite cell, but reduces the amount of Pb deposits on the muscle surface and connective tissue. This technique has been applied to determine the effects of denervation on the satellite cells of frog sartorius muscles. Four weeks after denervation, the number of satellite cells is essentially the same in both denervated muscles and the intact muscles of the contralateral side. However, denervation results in a subpopulation of satellite cells with altered shapes. They have elongated cytoplasmic processes which often branch. It is suggested that these supernumerary cytoplasmic processes represent an intermediate phase in the transition of satellite cells to myoblasts.

Animals↗
PubMed1995

Relationship between muscle myosin isoforms and contractile features in rabbit fast-twitch denervated muscle.

The effects of 8-day-old rabbit fast-twitch gastrocnemius denervation on the type of myosin isoforms and on contractile features (maximum velocity Vmax and contraction time (CT) of the muscle were followed between 15 and 60 days postnatal. The myosin isoforms and the Vmax and CT values of the denervated gastrocnemius displayed large changes during this period. These changes, which led at 2 months postnatal to a muscle displaying the properties of a slow-twitch muscle did not occur in synchrony: complete conversion to slow-type myosin isoforms occurred only at 60 days postnatal, whereas complete conversion to slow-twitch Vmax and CT values occurred as soon as 35 days postnatal. The results address a new question concerning the relationship between muscle myosin and contractile features.

Aging↗
PubMed1977

Is malignant hyperpyrexia muscle denervated?

To test the hypothesis that human muscular dystrophies may be secondary to denervation, the responses in vitro of muscle in human malignant hyperpyrexia to electrical and pharmacological stimuli have been compared with those of the denervated mouse soleus muscle. The results suggest that the muscle abnormality in malignant hyperpyrexia is different from that produced by denervation. This must cast doubt on the concept that other human muscular dystrophies are secondary to denervation.

Animals↗
PubMed2004

Electrical stimulation of denervated muscles.

PURPOSE: In this report, the 3-year rehabilitation of a 43-year-old male patient, who have pelvic 'open book' fracture, urinary bladder and urethral injury, lumbosacral plexus avulsion trauma and right lower extremity monoparesis is explained. METHOD: Rehabilitation of these injuries is absolutely essential to ensure optimal functional recovery. But the value of electrical stimulation for denervated muscle is not proven and its application to gain what may only be a small benefit is often not justified. CONCLUSIONS: With this case I emphasize that electrical stimulation and appropriate exercise programme in denervated protected muscle fibers activity for 3 years and rehabilitation should be continued.

Adult↗
PubMed2001

Increased neuromuscular activity reduces sprouting in partially denervated muscles.

The effects of increasing neural activity on sprouting remain unclear and controversial. In a rat model of partial denervation of skeletal muscles, we investigated the effect of neuromuscular activity on sprouting. Rat hindlimb muscles were partially denervated by avulsion of either L4 or L5 spinal root. Immediately after partial denervation, the rats were divided into three groups: (1) normal caged activity, (2) running exercise on wheels, 8 hr daily, and (3) functional electrical stimulation (FES) of sciatic nerves, 20 Hz for 8 hr daily. At 1 month, muscle unit (MU) enlargement was quantitated electrophysiologically and histochemically. MU twitch force was increased by four- to fivefold by partial denervation in extensively denervated tibialis anterior (TA) and medial gastrocnemius (MG) and by approximately twofold in moderately denervated plantaris (PL) and soleus (SOL). For the extensively denervated TA and MG muscles, MU enlargement, measured electrophysiologically, declined significantly after an average of 1757 +/- 310 m/d running exercise and daily FES for 1 month. The detrimental effects on MU enlargement were much less but significant in the moderately denervated PL and did not reach statistical significance in the moderately denervated SOL muscle. Histochemical evaluation of sprouting showed a reduction in the number of sprouts in the extensively denervated TA muscle, but not the moderately denervated PL and SOL muscles, by increased neuromuscular activity. Thus, increased neuromuscular activity is detrimental primarily in muscles that are extensively denervated, and the MUs are smaller than under conditions in which the muscles experience normal physiological levels of activation.

Animals↗
PubMed2003

Neuromuscular activity impairs axonal sprouting in partially denervated muscles by inhibiting bridge formation of perisynaptic Schwann cells.

Following partial denervation of rat hindlimb muscle, terminal Schwann cells extend processes from denervated endplates to induce and guide sprouting from the remaining intact axons. Increased neuromuscular activity significantly reduces motor unit enlargement and sprouting during the acute phase of sprouting. These findings led to the hypothesis that increased neuromuscular activity perturbs formation of Schwann cell bridges and thereby reduces sprouting. Adult rat tibialis anterior (TA) muscles were extensively denervated by avulsion of L4 spinal root and were immediately subjected to normal caged activity or running exercise (8 h daily) for 3, 7, 14, 21, and 28 days. Combined silver/cholinesterase histochemical staining revealed that the progressive reinnervation of denervated endplates by sprouts over a 1 month period in the extensively partially denervated TA muscles was completely abolished by increased neuromuscular activity. Immunohistochemical staining and triple immunofluorescence revealed that the increased neuromuscular activity did not perturb the production of Schwann cell processes, but prevented bridging between Schwann cell processes at innervated and denervated endplates. Our findings suggest that failure of Schwann cell processes to bridge between endplates accounts, at least in part, for the inhibitory effect of increased neuromuscular activity on sprouting.

Animals↗