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[Ca2+ metabolism in the sarcoplasmic reticulum of skeletal muscle after surgical denervation].

The ability of sarcoplasmic reticulum to regulate Ca(2+)-metabolism was studied at different terms after surgical denervation. The increase of Ca(2+)-ATP-ase activity, intensification of active Ca(2+)-accumulation under insignificant change of passive Ca(2+)-outflow from sarcoplasmic reticulum vesicles were observed 3-days after denervation. At more remote terms 14 days and 28 days after denervation all these parameters decreased except for Km: Ca(2+)-ATP activity and active Ca2+ accumulation decreased below normal level.

Animals↗

[Changes in the contractile properties of tonic muscle fibers following denervation].

After denervation, most of the tonic muscle fibers, contrary to intact fibers, cannot keep for a considerable time either the potassium (K) or the acetylcholine (A) contracture. The maximal mechanical tension in response to K and A in the denervated tonic fibers is lesser than in normal. The amplitude, speed of development, and latency (L) of the caffeine contracture remain practically the same. The speed of development of the K contracture sharply falls down and its L rather increases while the respective parameters of the A contracture practically do not change. The main changes developing after the denervation seem to occur in the processes controlling relaxation of the tonic fiber, as well as in the mechanisms for activation of K contracture while preserving the mechanisms for activation of A contracture.

Acetylcholine↗

Acetylcholine content and release in denervated or botulinum poisoned rat skeletal muscle.

1. The acetylcholine (ACh) content and spontaneous and evoked release of ACh in rat extensor digitorum longus (EDL) muscles were determined by pyrolysis-mass fragmentography. The determinations were made on muscles paralysed by local application of botulinum toxin (BoTx) type A, on unpoisoned muscles, surgically denervated or reinnervated muscles.2. The ACh content of unpoisoned control muscles was nearly uniform between animals and varied in the experimental series between 36 and 50 pmol. BoTx failed to affect the ACh content after 2 d of poisoning and caused a slight increase in content after 8 d. Surgical denervation reduced the ACh content within 24 h to less than 10% of innervated muscles and upon reinnervation the ACh content was restored. Following cholinesterase inhibition the ACh content of innervated and denervated muscles increased somewhat, about equally with time.3. Spontaneous release of ACh varied in normal innervated muscles between 40 and 100 fmol/min. In the presence of 25 mm-KCl the rate of release increased about fourfold. In BoTx poisoned muscles spontaneous release was reduced by up to 60% of control and high potassium failed to accelerate the release at 2 d after poisoning and caused only a small increase at 8 d. Denervated muscles released ACh at a rate which was less than 20% of control and it was not accelerated by high potassium.4. The results show that more than 90% of total ACh in the innervated EDL muscle is present in the nerve and its terminals. The remaining ACh is apparently formed and stored in the muscle tissue. BoTx caused a larger reduction in ACh release than can be accounted for by assuming a selective blockade of quantal release of transmitter. It suggests that BoTx has an inhibitory effect also on non-quantal ACh release.

Acetylcholine↗

Responses in the hypoglossal nucleus to delayed regeneration of the transected hypoglossal nerve, a quantitative ultrastructural study.

Electron micrographs of the left hypoglossal nucleus of adult male albino rats were quantitatively analyzed from 9-70 days after allowing the transected left hypoglossal nerve to regenerate after an 84 day delay. Delay was achieved by implanting the proximal stump into already innervated sternomastoid muscle, where no regeneration occurred. Regeneration was then allowed by denervating the sternomastoid. During the regenerative period the initially high number of abnormally electron dense perikarya and dendrites decreased to almost normal values, but no cell removal was seen. This suggested that the degenerate appearance of many profiles after prolonged prevention of regeneration, was reversible. The neuropil bouton and dendrite counts, and the numbers of synapsing boutons per dendrite, increased steadily to normal values from the low values of suppressed regeneration. Somatic bouton frequencies, even though already low, decreased further at 32 days, and later increased but not to normal values. The decrease at 32 days coincided with the loss of many subsurface cisterns, and dispersion of Nissl substance, all suggestive of chromatolysis. Later the subsurface cisterns and Nissl substance returned. It was suggested that the delay of complete recovery of somatic bouton frequencies might be because of lack of sensory information from the denervated muscle into which the hypoglossal nerve was regenerating, or because of abnormally low starting values for the recovery phase. Astrocyte (or, occasionally microglial) sheaths persisted along bouton-free perikaryal surfaces.

Animals↗

[Direct muscular neurotization].

The authors present a technique of direct muscular neurotization consisting of re-innervation by direct implantation of a nerve into one or several muscles in cases of avulsion of the nerve supplying the muscle or traumatic or surgical avulsion of the neural part of the muscle. The biological basis for this operation is that a denervated muscle can accept a new innervation by a foreign nerve even in an aneural zone because its sensitivity to acetylcholine is present throughout the muscle, while, in a normally innervated muscle, sensitivity to acetylcholine is confined to the motor endplates. The results of 47 cases are presented: 42 of them obtained good or very good results.

Animals↗

Metabolic characteristics of experimental free vascularized canine gracilis muscle transfers.

A canine gracilis model was used to study muscle energy metabolism and enzyme activities after free vascularized muscle transfer. Fifteen male mongrel dogs underwent orthotopic, free transfer of the left gracilis with microneurovascular anastomosis. After a minimum of 10 months' recovery, muscle biopsy specimens were obtained from the transfers and the contralateral controls and analyzed for relative fiber type areas and maximum activities of phosphorylase, hexokinase, phosphofructokinase, glycerol-3-phosphate dehydrogenase (GPDH), pyruvate kinase, lactate dehydrogenase, citrate synthase, succinate dehydrogenase, 3-hydroxyacyl coenzyme A dehydrogenase (HAD), and creatine phosphokinase. Biopsy specimens obtained before and after a 10 minute, 20-Hz contraction were analyzed for glucose, glycogen, glycolytic intermediates, phosphocreatine, total creatine, and adenine nucleotides (adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, inosine monophosphate, and inosine). There was no significant transfer versus control difference in type I relative fiber area (45 +/- 4 percent versus 44 +/- 3 percent). Total creatine was significantly reduced in the transferred muscles relative to control (83.1 +/- 3.0 mmol/kg versus 100.6 +/- 5.1 mmol/kg dry weight). Maximal activities of phosphorylase, pyruvate kinase, lactate dehydrogenase, citrate synthase, succinate dehydrogenase, HAD, and creatine phosphokinase were diminished in transfers relative to controls, although hexokinase activity was significantly higher in the freely transferred gracilis muscles. During the 20-Hz contraction, muscle transfers produced less force initially, although the force/time integral over the 10-minute stimulation was similar in transfers (277 +/- 25 N/g/second) and controls (272 +/- 24 N/g/second). The contraction was associated with significant glvcogen use and lactate accumulation in both transfers and controls, although this was less pronounced for the transfers. Glycolytic flux appeared muted in the transfers relative to controls. Significant, similar high-energy phosphagen reductions and inosine monophosphate accumulation were noted during the contraction in both groups. Contractile activity is associated with the expected pattern of muscle metabolite changes following free vascularized transfer, indicating the components of cellular energy metabolism are not qualitatively altered after microneurovascular muscle transfer. In contrast, quantitative differences suggest that free vascularized muscle transfer can be associated with a muscle enzyme profile consistent with deconditioning and the presence of denervated muscles fibers in the absence of fiber type profile changes.

Animals↗

Splinting for peripheral nerve injury in upper limb.

The prognosis and speed of peripheral nerve recovery depend very much on the level of injury, severity of injury, the surgical intervention and the subsequent rehabilitative process. Many high level injuries may take years or months for the affected peripheral nerve to recover. Prolonged muscle imbalance causes joint contractures and over-stretching of denervated muscles. Without proper care, hand function recovery may be limited even the nerve regenerated afterwards. During the nerve regeneration period, splinting is one of the most useful modality to minimise deformities, prevent joint contractures and substitute loss motor control. Proper splinting encourages early use of the injured hand in daily activities. There are different types of splinting design for median nerve palsy, ulnar nerve palsy and radial nerve palsy. Dynamic splinting techniques are frequently employed to allow early prehension activities. Other therapeutic techniques, including pressure garment and sensory re-education are useful to enhance better functional return after nerve repair.

Arm Injuries↗

Denervation in hemiplegic muscles.

This study examined the frequency of denervation activity in hemiplegic muscles in relation to the size and location of the central lesion. We studied 20 patients, 14 with major unilateral cerebral infarctions in the middle cerebral or internal carotid territories; four with a single lacune in the pons, internal capsule, or thalamus; and two with precentral infarcts. Using somatosensory evoked potentials, motor conduction studies, and assessments of conduction across the plexus and roots, we detected no conduction abnormalities on the affected side. Fibrillation was common in both groups, especially in distal and intermediate muscles. The distribution of the fibrillation and the normal conduction studies suggested that trauma of peripheral nerves was not a factor. Although the normal conduction studies and pattern of fibrillation activity do not exclude peripheral nerve trauma as the cause of the fibrillation, we suggest that transsynaptic degeneration is a reasonable alternative explanation.

Action Potentials↗

Distribution of motor nerve sproutings in the mouse gastrocnemius muscle after partial denervation.

The distribution and frequency of motor nerve sproutings in the mouse gastrocnemius muscle were examined after a partial denervation. Terminal and nodal sproutings could be seen for a period of 7-28 days after transecting one of the two muscular nerves in the medial head. They grew out not only from the endplates and terminal nerves, but also from the preterminal nerve regions of both the intra- and extramuscular nerve. The terminal sproutings originated from endplates close to the denervated portion and subsequently they were seen farther away in the medial head. Upon regeneration of the originally transected nerve the terminal sproutings were withdrawn in the reverse sequence, i.e., first from the region farthest from the denervated area and last from the region adjacent to the denervation. The frequency of terminal sproutings was constant in the area immediately bordering the denervated portion (11-15% of the total number of endplates). In other areas more distant from the denervated portion, however, it showed a gradient which varied with the time course. The higher the frequency of terminal sproutings, the more actively each endplate produced two or three terminal sproutings without significant difference in length.

Animals↗

Effect of denervation on cyclic amp regulating enzymes in the rat gastrocnemius muscle.

The activities of cyclic AMP regulatory enzymes (adenylate cyclase and phosphodiesterase) were studied in rat gastrocnemius muscle after denervation. Basal adenylate cyclase activity in whole homogenate increased progressively from the 3rd day reaching values more than 7-fold those of the controls 30 days after denervation. No significant differences between the contralateral and denervated muscle were detected as regards adenylate cyclase activation by catecholamines or NaF. Carbamylcholine was with effect. Cyclic AMP phosphodiesterase activity increased significantly at 3 days after denervation as was still significantly elevated at 60 days.

3',5'-Cyclic-AMP Phosphodiesterases↗

Sodium channel mRNAs at the neuromuscular junction: distinct patterns of accumulation and effects of muscle activity.

Voltage-gated sodium channels (VGSCs) are highly concentrated at the neuromuscular junction (NMJ) in mammalian skeletal muscle. Here we test the hypothesis that local upregulation of mRNA contributes to this accumulation. We designed radiolabeled antisense RNA probes, specific for the "adult" Na(V)1.4 and "fetal" Na(V)1.5 isoforms of VGSC in mammalian skeletal muscle, and used them in in situ hybridization studies of rat soleus muscles. Na(V)1.4 mRNA is present throughout normal adult muscles but is highly concentrated at the NMJ, in which the amount per myonucleus is more than eightfold greater than away from the NMJ. Na(V)1.5 mRNA is undetectable in innervated muscles but is dramatically upregulated by denervation. In muscles denervated for 1 week, both Na(V)1.4 and Na(V)1.5 mRNAs are present throughout the muscle, and both are concentrated at the NMJ. No Na(V)1.5 mRNA was detectable in denervated muscles stimulated electrically for 1 week in vivo. Neither denervation nor stimulation had any significant effect on the level or distribution of Na(V)1.4 mRNA. We conclude that factors, probably derived from the nerve, lead to the increased concentration of VGSC mRNAs at the NMJ. In addition, the expression of Na(V)1.5 mRNA is downregulated by muscle activity, both at the NMJ and away from it.

Animals↗

Carnitine and acyltransferase in experimental neurogenic atrophies: changes with treatment.

Carnitine level and carnitine palmityl transferase (CPT) activity were investigated in muscles of patients with infantile and juvenile spinal muscular atrophy and polyneuropathies. A significant decrease of both carnitine and CPT was found in the infantile spinal muscular atrophy, but not in the other neurogenic muscle atrophies. These findings were compared with the experimental effect of denervation and reinnervation upon the lipid metabolism in soleus and extensor digitorum longus (EDL) of adult and newborn rats. Twenty-one days after denervation free and total carnitine decreased significantly in both EDL (P less than 0.001) and soleus (P less than 0.05) of adult animals. CPT activity was significantly decreased in the soleus 50 days after denervation (P less than 0.005). Long-term reinnervation restored the level of carnitine fraction and CPT activity. L-carnitine treatment for 21 days restored the level of free carnitine to normal in the soleus of denervated adult animals. Denervation in newborn rats influenced carnitine concentration in soleus and EDL to a lesser extent; the treatment with L-carnitine raised short-chain acylcarnitines in denervated muscles, while reinnervation restored carnitine level within 50 days.

Acyltransferases↗

Differences in the interaction of acetylcholine receptor antibodies with receptor from normal, denervated and myasthenic human muscle.

The interaction of acetylcholine receptor antibodies with different kinds of human skeletal muscle receptor was investigated. The reaction of most receptor antibodies was strongest with receptor from a patient with myasthenia gravis and with receptor from denervated muscle. Results obtained with these receptors were well correlated. The binding of most receptor antibodies to receptor from functionally normal muscle was much weaker and also qualitatively different. In a few patients with moderate and severe clinical symptoms the existence of acetylcholine receptor antibodies was revealed only by their reaction with receptor from myasthenic muscle. One patient had a clone of receptor antibodies reacting exclusively with receptor from her own muscle.

Aged↗

Regulation of translation factors during hindlimb unloading and denervation of skeletal muscle in rats.

In the rat, denervation and hindlimb unloading are two commonly employed models used to study skeletal muscle atrophy. In these models, muscle atrophy is generally produced by a decrease in protein synthesis and an increase in protein degradation. The decrease in protein synthesis has been suggested to occur by an inhibition at the level of protein translation. To better characterize the regulation of protein translation, we investigated the changes that occur in various translation initiation and elongation factors. We demonstrated that both hindlimb unloading and denervation produce alterations in the phosphorylation and/or total amount of the 70-kDa ribosomal S6 kinase, eukaryotic initiation factor 2 alpha-subunit, and eukaryotic elongation factor 2. Our findings indicate that the regulation of these protein translation factors differs between the models of atrophy studied and between the muscles evaluated (e.g., soleus vs. extensor digitorum longus).

Animals↗

Expression of RNA transcripts for the postsynaptic 43 kDa protein in innervated and denervated rat skeletal muscle.

A cDNA clone encoding the mouse muscle postsynaptic 43 kDa protein was isolated and sequenced. The amino acid sequence of this protein, which is closely associated with nicotinic acetylcholine receptors at Torpedo electrocyte and vertebrate skeletal muscle synapses, is very similar in different species. A cysteine-rich region homologous to part of the regulatory domain of protein kinase C may be important in interactions of this protein with the lipid bilayer. RNA transcripts for the 43 kDa protein increase only 2-3 fold after denervation of rat skeletal muscle, in sharp contrast to the alpha-subunit of the muscle nicotinic receptor which increases more than 30-fold. Thus, the expression of these two proteins is regulated by different mechanisms.

Amino Acid Sequence↗

[Clinical significance and changes of fibrillation potential amplitude following denervation of laryngeal muscles].

OBJECTIVE: To evaluate the value of clinical application of examination of fibrillation potential amplitude of laryngeal electromyogram. METHOD: Fifty-four patients (65 sides) with recurrent laryngeal nerve paralysis were examined and only the maximum fibrillation potential amplitudes were recorded in the posterior cricoarytenoid muscles and thyroarytenoid muscles. RESULT: There was no significant difference between sexes, ages and sides. The fibrillation potential amplitude was maximum at 3 to 4 months after denervation of the posterior cricoarytenoid muscle or at 5 to 6 months after denervation of the thyroarytenoid muscle and still remained at certain level for years in some patients. Significant difference was showed in complete and incomplete nerve injuries groups, but changes of amplitude with time in incomplete nerve injuries group were small. CONCLUSION: The changes of fibrillation potential amplitude might indicate that the changes from muscle denervation was still reversible and might be more accurate than traditional method of examination.

Adult↗

Immunoperoxidase localization of alpha bungarotoxin: a new approach to myasthenia gravis.

The IPBT method has made it possible to precisely visualize the AChR. Normal distribution of AChR is at the peaks of the postjunctional folds of the muscle sarcolemmal membrane with a small amount present on the axonal tip as well. Denervated muscle fibers have extrajunctional AChR. In MG, there are also denervated-appearing fibers but these do not have extrajuctional AChR with the IPBT stain. To explain this, we have been able to demonstrate a serum factor capable of blocking the binding of alpha-BuTx to the AChR and have shown for the first time that this factor is capable of acting at the neuromuscular junction itself. This blocking factor may play a major role in causing the weakness of MG.

Acetylcholine↗