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PubMed1983

[Effect of thyroxine on the phenotype of intact and denervated skeletal muscles in the guinea pig].

Denervation of the guinea-pig fast (m. plantaris) and slow (m. soleus) muscles, administration of alpha-thyroxine (T4) to animals followed by denervation and T4 injection entail changes in the relative content of different types of muscle fibers. The slow muscle shows fast muscle fibers after T4 injection, denervation and T4 injection. Moreover, myosin isolated from the m. soleus shows the precipitation band with antibodies to fast muscle myosin. These data attest to partial reprogramming of myosin synthesis by the m. soleus, pointing to the role played by thyroxine in myosin synthesis regulation.

Animals↗
PubMed1977

Denervated skeletal muscle fibers develop discrete patches of high acetylcholine receptor density.

Denervated skeletal muscle fibers of mice develop discrete patches of high acetylcholine receptor density. The patches vary in size from less than 1 micrometer up to 30 micrometers, depending on the muscle and the period of denervation. Within the patches the acetylcholine receptor density is some 20 times greater than elsewhere along the muscle fiber and probably approaches that in the subsynaptic membrane.

Acetylcholine↗
PubMed1990

Ultraterminal sprouting in innervated and partially denervated adult and aged rat muscle.

End plates from the extensor digitorum longus, soleus and diaphragm muscles of adult (10-month-old) and aged (25-month-old) rats were examined to determine whether aging affects the frequency of occurrence of ultraterminal sprouting, a form of terminal sprouting commonly associated with muscle denervation. There was a significant increase in the fraction of end plates exhibiting ultraterminal sprouts in the extensor digitorum longus muscle only; the number increased from 5.5% in 10-month-old animals to 23.0% in 25-month-old animals. To see whether age also influences the sprouting response to denervation, extensor digitorum longus muscles from 10-and 25-month-old animals were partially denervated by severing spinal nerve L5, and end plates were examined 4-7 and 10-14 days following denervation. Denervation induced a more profound increase in the percentage of end plates exhibiting ultraterminal sprouting in the 10- compared with 25-month-old animals. However, sprouting remained significantly greater in the 25- compared with the 10-month-old animals. Furthermore, following denervation the average ultraterminal sprout length was significantly greater in the 25- compared with the 10-month-old animals. There was no correlation between the extent of muscle denervation and the percentage of end plates exhibiting ultraterminal sprouts. End plates with ultraterminal sprouts had larger areas and contained fewer nerve terminal branches than end plates without these sprouts. It is suggested that the limited response to partial denervation in the 25-month-old extensor digitorum longus muscles may indicate that the aged extensor digitorum longus has already approached the maximum capacity for sprouting.

Animals↗
PubMed1978

Active state properties of denervated and immobilized muscle: comparison with dystrophic muscle.

Possible roles of neurotrophic mechanisms and muscle activity in the contractile abnormalities of muscular dystrophy were studied by comparing human dystrophic muscle to denervated and immobilized muscle. As evident in denervated muscle from the decreased acceleration of twitch development (decreased active state intensity of shortening), and isoproterenol-induced decrease of twitch with shortened decay of the active state, part of the abnormality in the subcellular calcium transport system in dystrophic muscle seems to be influenced by disordered neural regulation. Other active state abnormalities relating to activation processes and contractile proteins in dystrophic muscle were also demonstrated in both denervated and immobilized muscle, with some being more marked in immobilized muscle. The findings indicate that a neurogenic hypothesis cannot entirely explain the pathogenesis of progressive muscular dystrophy.

Adult↗
PubMed1980

Partial denervation in inactive muscle effects innervated and denervated fibres equally.

Possible causal factors of denervation-induced changes in muscle include inactivity, products of nerve degeneration and lack of a nerve-borne trophic agent. We now show that if the innervated fibres in a partially denervated rat muscle are rendered inactive, they undergo a reaction as intense as that of the denervated fibres. This provides further support for the view that the effects of denervation on the extrajunctional muscle membrane result from a combination of muscle inactivity and of nerve breakdown products acting diffusely throughout the muscle.

Action Potentials↗
PubMed1987

Neurite outgrowth on cryostat sections of innervated and denervated skeletal muscle.

To localize factors that guide axons reinnervating skeletal muscle, we cultured ciliary ganglion neurons on cryostat sections of innervated and denervated adult muscle. Neurons extended neurites on sections of muscle (and several other tissues), generally in close apposition to sectioned cell surfaces. Average neurite length was greater on sections of denervated than on sections of innervated muscle, supporting the existence of functionally important differences between innervated and denervated muscle fiber surfaces. Furthermore, outgrowth was greater on sections of denervated muscle cut from endplate-rich regions than on sections from endplate-free regions, suggesting that a neurite outgrowth-promoting factor is concentrated near synapses. Finally, 80% of the neurites that contacted original synaptic sites (which are known to be preferentially reinnervated by regenerating axons in vivo) terminated precisely at those contacts, thereby demonstrating a specific response to components concentrated at endplates. Together, these results support the hypothesis that denervated muscles use cell surface (membrane and matrix) molecules to inform regenerating axons of their state of innervation and proximity to synaptic sites.

Animals↗
PubMed1975

Desensitization in the innervated and in the chronically denervated soleus muscle of the mouse.

1 A study was made of desensitization in chronically denervated and in normally innervated mouse soleus muscles. 2 Very high concentrations of acetylcholine produced small contractions of the innervated muscles; these were reduced in size when the addition was repeated 1 min after wash-out. 3 Desensitization in innervated muscles was receptor specific: contractions in response to KCl and caffeine were not reduced following acetylcholine, nor did KCl produce desensitization. 4 In chronically denervated muscles non-specific desensitization was observed if acetylcholine was added in the presence of low concentrations of acetylcholine or carbachol. Contractions to KCl but not to caffeine were reduced. KCl produced a similar kind of desensitization. 5 After washing out moderate or supramaximal concentrations from the chronically denervated muscle no desensitization was observed. However, 1 min after washout of very high concentrations the muscle was non-specifically desensitized. 6 It is concluded that relatively high concentrations of acetylcholine are required to produce specific desensitization in the soleus muscle. Chronically denervated muscles, being supersensitive, show non-specific loss of sensitivity with concentrations of acetylcholine too low to produce specific desensitization.

Acetylcholine↗
PubMed1986

An ultrastructural study of the segmental uptake of horseradish peroxidase in the endplate region of denervated skeletal muscle fibres.

The segmental uptake of horseradish peroxidase (HRP) in the endplate region of denervated skeletal muscle fibres has been studied ultrastructurally using a method for selecting single muscle fibres with high segmental peroxidase staining from denervated mouse tibialis anterior muscle. Segments containing large peroxidase positive phagosomes could already be seen 10-15 min after i.v. injection of HRP. Such segments were still present 24 h after HRP injection. The localization of phagosomes, deep in the fibres rather than immediately under the sarcolemma, suggests that the uptake occurs from t-tubuli. Vivid proliferation of t-tubuli, consisting of vesiculation, enlargement and encircling of cytoplasmic components, was also observed. The HRP accumulates in phagosomes of varying size and shape. Similar membrane-limited bodies without or with very weak peroxidase staining were also observed. The peroxidase-positive phagosomes participate in autophagic processes as suggested by their content of undegraded cellular material. Golgi profiles, which occurred deep in the muscle fibres, and enlarged components of the sarcoplasmic reticulum were frequently encountered in the segments. Myofibrillar degeneration occurs in the segments and progresses with time after denervation. The described segments may be related to the increased membrane turnover in denervated muscle fibres and/or they may be related to processes aimed at establishing new synaptic contacts.

Animals↗
PubMed1994

Blood flow, mitochondria, and performance in skeletal muscle after denervation and reinnervation.

Tibialis anterior (TA) muscles of rats underwent bilateral peroneal nerve crush (NC) or denervation (D) and were compared with sham-operated (SO) animals to determine the effect of reinnervation on blood flow, mitochondria, metabolites, and muscle performance. After surgery, animals were left for 2, 7, 21, or 42 days (NC and SO groups) or 2, 7, or 21 days (D group; n = 7-11.day-1.group-1), after which TA muscles were stimulated in situ at 1 Hz. alpha-Motoneuron reinnervation of muscle was complete 21 days after NC. Blood flow increased 10-fold above SO values in nonstimulated TA muscle 7 days after NC and D (P < 0.05). By 21 days, blood flow to nonstimulated TA muscle in NC animals returned to SO values but remained elevated (P < 0.05) in D muscle. Thus restoration of neural control of blood flow to resting muscle likely occurred by 21 days post-NC. Blood flow to stimulated muscle was not affected by NC or D, indicating the probable importance of metabolic factors in regulating blood flow during 1-Hz contractions. Cytochrome-c oxidase activity decreased (P < 0.05) below SO values 7 days after NC and D. By 21 days, cytochrome-c oxidase activity in TA muscles of NC animals returned to SO values, while values in denervated TA muscle continued to decrease. Despite these changes, endurance performance of TA muscle was not affected by D or NC at any time. These results suggest that reinnervation processes controlling blood flow and muscle function occur along similar time courses and that muscle blood flow is more closely related to endurance performance than is muscle oxidative capacity under these contraction conditions.

Animals↗
PubMed1982

Identification of an intracellular postsynaptic antigen at the frog neuromuscular junction.

A layer of amorphous, electron-dense material is situated at the cytoplasmic surface of the postsynaptic membrane of vertebrate neuromuscular synapses. The function of this structure is not clear, but its location suggests that it may have an important role in the formation and/or maintenance of the synapse. This paper demonstrates that a monoclonal antibody raised against antigens from Torpedo electric organ binds to an intracellular, postsynaptic protein at the frog neuromuscular synapse. Indirect immunofluorescence on frozen sections of frog muscle was used to demonstrate that the antigen is concentrated at synaptic sites in normal muscle. In denervated muscle, the antigen remains concentrated at synaptic sites, but is also present at extrasynaptic regions of denervated myofibers. The antigen cannot be labeled in intact, whole muscle, but only in whole muscle that has been permeabilized with nonionic detergents. The antibody staining pattern in Triton X-100-permeabilized whole-mounts of the frog neuromuscular synapse is arranged in elongate, arborized areas which are characteristic of the frog neuromuscular synapse. The stained areas are striated and the striations occur with a periodicity that corresponds to the regular folding of the postsynaptic membrane. Immunoferritin labeling of fixed, saponin-permeabilized muscle demonstrates that the antigen is associated with amorphous material that is situated between the postsynaptic membrane and an underlying layer of intermediate filaments. The antigen, solubilized from membrane and an underlying layer of intermediate filaments. The antigen, solubilized from Torpedo electric organ by high ionic strength, was identified by antibody binding to nitrocellulose replicas of SDS gels of Torpedo tissue. In Torpedo tissue, the antibody binds to a single protein band at 51,000 daltons (51 kd). The 51-kd protein shares an antigenic determinant with intermediate filament proteins, since a monoclonal antibody to all intermediate filaments reacts with the same 51-kd protein. The monoclonal antibody also reacts with a 55-kd protein in frog skin which is localized to the perinuclear region of the epithelial cells.

Animals↗
PubMed1985

Can cells extruded from denervated skeletal muscle become circulating potential myoblasts? Implications of 3H-thymidine reutilization in regenerating muscle.

The hypothesis that satellite cells which leave denervated skeletal muscle might become circulating potential myoblasts which could participate in myogenesis in distant sites in the body has been tested. Sixteen mice had one hindlimb denervated and were given 7 daily injections of 3H-thymidine (3H-Tdr). One day later extensor digitorum longus muscle isografts from unlabelled mice were inserted into each hindlimb. As controls, the procedure was repeated in 6 non-denervated labelled mice. Fourteen days after their insertion, isografts in denervated mice contained many labelled myotubes with a labelling index of 55 +/- 4% (mean +/- SEM). In the control isografts in non-denervated ice, 38 +/- 4% of myotube nuclei were labelled. The results show that either labelled cells, or 3H-Tdr, had transferred from the host to isografts in both cases. The probability of 3H-Tdr reutilization was demonstrated in regenerating livers of 8 similarly labelled mice, where 34 +/- 3% hepatocytes adjacent to crush lesions were labelled after 14 days. This conclusion was reached because only 2-3% of normal hepatocytes incorporate 3H-Tdr under these conditions and this population is inadequate to provide sufficient labelled precursor cells for the large numbers of labelled regenerated hepatocytes. Therefore, it was concluded that 3H-Tdr reutilization is the most likely explanation for labelled myotube nuclei in the muscle isografts (rather than movement of labelled precursor cells), and that additional label for reutilization had been derived from breakdown of labelled cells in denervated muscle. The data do not support the hypothesis of a circulating precursor for skeletal muscle cells.

Animals↗
PubMed1979

Autoradiographic visualization of beta-adrenergic receptors in normal and denervated skeletal muscle.

Autoradiographic localization of beta-adrenergic receptors in rat skeletal muscle in vivo was achieved utilizing [125I]-iodohydroxybenzylpindolol, a potent beta-adrenergic blocker with high affinity and specificity for those receptors. In normal muscle the beta-adrenergic receptors were localized mainly to blood vessels, arterioles greater than venules, with much less concentration of grains over the fascicles of muscle fibers. One week after denervation there was an increase in binding both to blood vessels and muscle fibers, more so in soleus and gactrocnemius than in extensor digitorum longus. While these results parallel in vitro biochemical studies, they dictate caution when inferring cellular localization of beta-adrenergic receptors (and other molecules) solely on the basis of biochemical techniques applied to subcellular fractions of whole-organ homogenates.

Animals↗
PubMed1989

[Activation of the fast calcium input in the denervated smooth muscle of the cat nictitating membrane].

The excitation and contraction features of innervated and sympathetically denervated smooth muscle strips from cat's nictitating membrane have been studied by single sucrose gap arrangement. Increasing of smooth muscle cells sensitivity to drugs were accompanied by elevation of membrane response and the ability to generation of action potentials. Action potentials have been induced by agonists or high potassium concentration in external solution and spontaneously. In innervated muscle action potentials have been evoked as a result of depolarization by high potassium concentration of TEA blockade of potassium conductance. Induced and spontaneously generated action potentials were blocked by organic and inorganic antagonists of potential dependent Ca++ channels. In Ca-free solution action potentials were absent but might be supported by Ba++. Decrease of Na+ had no effect on smooth muscle excitability. It is supposed that activation of potential depended Ca++ channels in smooth muscle cells with pharmaco-mechanical coupling are under influence of sympathetic nerves.

Action Potentials↗
PubMed2004

[The study on the regeneration of skeletal muscles after denervation].

OBJECTIVE: To study the degeneration and regeneration of skeletal muscle after denervation. METHODS: Denervation was carried out in gastrocnemius muscles in 30 adult BALB/C mice by cutting the sciatic nerve. The gastrocnemius muscles were removed at 1, 2, 4, 8, 12, 16 weeks after denervation, respectively. Specimens were processed for histological study and immunohistochemical technique. RESULTS: Muscle fiber atrophy followed by degerneration and regeneration was observed in the early period of denervation. Fusion of the regenerated muscle cells with each other followed by degeneration of the cells and growth of fibro-connective tissue were observed in the later stage. The expression of myoglobin and actin decreased in 1-4 weeks after denervation. The postive expression of the proteins was observed in some 8 weeks' cells and in many degenerated 12-14 weeks' muscle cells. CONCLUSION: Degeneration and regeneration may coexisted in the denervated muscles. The regenerated muscle cells can't fully develop due to the deficit of nerve regulation and degenerate again. The regenerated muscle cells will melt each other and can't develop to mature muscle fiber in the later stage.

Animals↗
PubMed1990

Potentiation of the halothane-cooling contractures of mammalian muscles by denervation.

Denervation potentiated the cooling-induced contractures and the halothane-cooling contractures of isolated extensor digitorum longus and soleus muscles of the mouse. These effects were more striking in extensor digitorum longus than in soleus muscles. Significant increases in the peak amplitudes of the halothane-cooling contractures of both muscles and of the cooling contractures of soleus muscle were observed within 2 and 7 days of denervation. The potentiation of the contractures persisted for 90 days, the period of this study. Denervation (greater than 2 days) endowed extensor digitorum longus with the ability to generate cooling contractures in the absence of halothane. The rate of tension development of cooling-induced contractures in the absence or presence of halothane was significantly greater in denervated (2-90 days) than in innervated muscles. Denervation also reduced the effectiveness of procaine in inhibiting the halothane-cooling contractures. It is proposed that the potentiation of cooling-induced contractures in denervated muscles results primarily from an increase in the rate of efflux and in the quantity of Ca2+ released from the sarcoplasmic reticulum, upon cooling and (or) when challenged with halothane.

Animals↗
PubMed1995

Cloning of the alpha component of the chick ciliary neurotrophic factor receptor: developmental expression and down-regulation in denervated skeletal muscle.

A full-length cDNA clone encoding for the chick CNTFR alpha (alpha component of the ciliary neurotrophic factor receptor) was isolated by screening an embryonic day 13 chick brain cDNA library with a rat CNTFR alpha probe. The isolated cDNA clone contained a approximately 2-kb insert with an open reading frame of 362 amino acids. The identification of this clone as chick CNTFR alpha was based on the homology in amino acid sequence (approximately 70%) with the rat and human CNTFR alpha. Hydropathy analysis revealed that the chick CNTFR alpha contains a hydrophobic region at the amino terminus that is typical of secretory signal peptides, as well as a hydrophobic region at the carboxyl terminus that is characteristic of glycosylphosphatidylinositol-linked proteins. The expression of chick CNTFR alpha was developmentally regulated and was widely distributed in neural tissues, such as brain and spinal cord. In the periphery, chick CNTFR alpha transcript was expressed at high levels in the skeletal muscle and was only barely detectable in the liver. Unexpectedly, the expression of chick CNTFR alpha mRNA in skeletal muscle was decreased by approximately 10-fold at 1.5 days after denervation. This is in sharp contrast to the result previously obtained with CNTFR alpha in denervated rat muscle.

Aging↗
PubMed1998

[Embryonic motoneurons transplanted into denervated skeletal muscles].

OBJECTIVE: To observe the living situation of homografting embryonic motoneurons in denervated skeletal muscles. METHODS: We transplanted dissociated ventral spinal cord cells from 12 embryonic days rats prelabeled with 5-bromo-2-deoxyuridine(Brdur) into denervated skeletal muscles of adult rats. The denervated skeletal muscles were collected at 9 and 22 weeks respectively, and Nissl, acetylcholinesterases immunocytochemistry and ATPase staining were used to show the transplanted embryonic motoneurons and the morphography of muscle fibers. RESULTS: Transplanted embryonic motoneurons could survive, extend out long axons, and form neuromuscular junctions with muscle fibers. Type grouping fibers with bigger transverse area were also observed in the experimental group. Muscle fibers were much smaller and did not form fiber type grouping in the control group. CONCLUSION: Homografting embryonic motoneurons could survive well and reinnervate the denervated skeletal muscles.

Animals↗
PubMed1992

Slow-to-fast transformation of denervated soleus muscle of the rat, in the presence of an antifibrillatory drug.

The myofibrillar changes of rat denervated soleus muscle were studied in the presence and in the absence of an antifibrillatory drug. After bilateral sciaticotomy, a concentrated solution of procainamide hydrochloride was steadily released, by way of a miniosmotic pump, in the space between the soleus and the gastrocnemius muscles of one leg. Fibrillation activity of soleus muscles was checked electromyografically at 3- to 5-day intervals. On the 21st day following denervation the muscles were excised, stained for adenosine triphosphatase activity and analysed for myosin heavy chain (MHC) isoforms. In the denervated-procainamide-treated muscles fibrillation was consistently (-75% on average) depressed in comparison to the contralateral denervated muscles. Type 1 (slow) fibres and MHC isoform were also significantly reduced, to the advantage of type 2A (fast) fibres and MHC isoform. The results support the view that denervation inactivity, like other kinds of muscle inactivity, favours the expression of fast type myofibrillar isoforms, and that this effect is counteracted, at least partially, by the spontaneous activity of the denervated muscle.

Animals↗