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PubMed2002

Effect of muscle electrostimulation on afferent activities from tibialis anterior muscle after nerve repair by self-anastomosis.

Numerous previous studies were devoted to the regeneration of motoneurons toward a denervated muscle after nerve repair by self-anastomosis but, to date, few investigations have evaluated the regeneration of sensory muscle endings. In a previous electrophysiological study (Decherchi et al., 2001) we showed that the functional characteristics of tibialis anterior muscle afferents are affected after self-anastomosis of the peroneal nerve even when the neuromuscular preparation was not chronically stimulated. The present study examines the regeneration of groups I-II (mechanosensitive) and groups III-IV (metabosensitive) muscle afferents by evaluating the recovery of their response to different test agents after self-anastomosis combined or not with chronic muscle stimulation for a 10-weeks period. We compared five groups of rats: C, control; L, nerve lesion without suture; LS, nerve lesion with suture; LSE(m): nerve lesion plus chronic muscle stimulation with a monophasic rectangular current; and LSE(b): nerve lesion plus chronic stimulation with a biphasic current with modulations of pulse duration and frequency, eliciting a pattern of activity resembling that delivered by the nerve to the muscle. Compared to the control group, (1) muscle kept only its original weight in the LSE(b) group, (2) in the LS group the response curve to tendon vibration was shifted toward the highest mechanical frequencies and the response of groups III-IV afferents after fatiguing muscle stimulation lowered, (3) in the LSE(m) group, the pattern of activation of mechanoreceptors by tendon vibrations was altered as in the LS group, and the response of metabosensitive afferents to KCl injections was markedly reduced, (4) in the LSE(b) group, the response to tendon vibration was not modified and the activation of metabosensitive units by increased extracellular potassium chloride concentration was conserved. Both LSE(b) and LSE(m) conditions were ineffective to maintain the post muscle stimulation activation of metabosensitive units as well as their activation by injected lactic acid solutions. Our data indicate that chronic muscle electrostimulation partially favors the recovery of mechano- and metabosensitivity in a denervated muscle and that biphasic modulated currents seem to provide better results.

Action Potentials↗

PubMed1996

Sarco(endo)plasmic reticulum Ca2+ pump and metabolic enzyme expression in rabbit fast-type and slow-type denervated skeletal muscles. A time course study.

Recent reports by d'Albis et al. have shown that denervation of 8-day-old rabbit fast-twitch muscle (gastrocnemius) leads to the transformation of the muscle towards a slow phenotype but the changes towards slow-type myosin isoforms and contractile properties of the muscle were temporally uncoordinated. We analyzed the time course of the effects of denervation of the gastrocnemius on the expression of the sarcoplasmic reticulum calcium pump isoforms (SERCA) and on the metabolic state of the muscle. Northern-blot analysis showed a rapid loss of the fast Ca2+ pump isoform (SERCA 1) mRNA from the denervated gastrocnemius which became of the oxidative type. The changes observed were complete as early as 35 days post-natal, i.e at the time when changes in contractile properties were previously observed. Denervation of the slow-twitch soleus led to a 50% decrease in the level of the slow Ca2+ pump isoform (SERCA 2) mRNA and was without effect on the metabolic state of the muscle. These findings extend previous results suggesting that in rabbit, continuous innervation is required for differentiation of fast-twitch muscles but is not an absolute requirement for differentiation of the slow-twitch muscle.

Animals↗

PubMed1989

Membrane ionic conductances in normal and denervated skeletal muscle of the rat during development.

The development of membrane ionic conductances of rat extensor digitorum longus (EDL) muscle fibers was studied in vitro using intracellular recordings. At 7-8 days after birth, the potassium conductance (GK) dominated the total membrane conductance while the chloride conductance (GCl) was very low. A rapid increase of GCl towards adult values was observed after few days (12-14 day old rats), whereas GK did not decrease up to day 23. Denervation at 7-8 days after birth suppressed the maturation of the electrical parameters measured, and 15 days after the nerve crush, GCl was just detectable. These results suggest that the maturation of the electrical properties, and in particular that of the resting chloride conductance in mammalian striated muscle fibers, occurs during the first weeks of postnatal life and is dependent on innervation.

Animals↗

PubMed1978

Changes in the satellite cells of growing muscle following denervation.

Satellite cells exhibit a number of distinct morphological changes after denervation which appear to be a direct response to nerve section or altered functional state of the muscle fibers. These changes appear generally related to increased movement and overall activation of the cells. After longer periods of denervation many satellite cells appear to separate from their fibers and become free cells in the interstitial space. It is proposed that this mechanism provides a cellular source for the small-diameter, immature fibers dispersed throughout the muscle after two to three weeks. Although neo-formation of myofibers appears to be a feature of denervated growing muscle, an increase in total fiber population was not observed. It is suggested that in the absence of a viable nerve supply the new fibers degenerate.

Animals↗

PubMed1991

SkM2, a Na+ channel cDNA clone from denervated skeletal muscle, encodes a tetrodotoxin-insensitive Na+ channel.

Approximately one third of the Na+ channels expressed in denervated or developing skeletal muscle are tetrodotoxin (TTX) insensitive, with a Kd for channel blockade of approximately 1 microM, similar to that found for cardiac Na+ channels. We have recently reported the cloning of a putative Na+ channel subtype that is characteristic of denervated and developing skeletal muscle (SkM2), the deduced amino acid sequence of which is identical to that of a Na+ channel cDNA isolated from heart. We have now examined the functional properties of SkM2 Na+ channels after expression in Xenopus oocytes. We found that the efficiency of expression of constructs containing the SkM2 clone was strongly dependent on the amount of 5'-untranslated region (5'UTR) included. Constructs containing a 206-nucleotide 5'UTR were expressed poorly, whereas constructs from which most of the 5'UTR was removed were expressed well. The channels showed rapid voltage-dependent activation and inactivation. In addition, SkM2 Na+ channels were insensitive to low concentrations of TTX but were ultimately blocked by this toxin, with a Kd of 1.9 microM. The TTX block exhibited use dependence. Finally, SkM2 Na+ channels were not blocked by 100 nM mu-conotoxin, which blocks Na+ channels in innervated skeletal muscle in the low nanomolar concentration range. These data indicate that SkM2 Na+ channels are the TTX-insensitive Na+ channels found in denervated or developing skeletal muscle and are identical to the TTX-insensitive Na+ channels from heart.

Animals↗

PubMed2002

[Effect of trigeminus nerve on facialis-denervated facial muscle atrophy].

In the present study we made out an animal model on rabbit whose trigeminus and facialis nerves were simultaneously or only the latter one was severed. The pathological changes in facial muscle atrophy under different nerve injuries were investigated. The degeneration of contractile proteins of upper lip muscle -- myosin and actin was observed. In addition, we also examined the ultrastructural changes in the muscle atrophy in the two above-mentioned nerve injury cases. We observed that the intact trigeminus nerve could delay and lighten the atrophy of facialis-denervated facial muscle and attenuate the degeneration of myosin and actin, as well as decrease the increment of collagen and maintain the ultrastructure of the thick and thin muscle filaments. These results may provide the possibility of improvement of clinical treatment for facial muscle palsy.

Animals↗

PubMed1977

Increased histaminase activity in the atrophic muscle of denervated frog.

Sciatic denervation for 1 month in the frog Rana hexadactyla resulted in progressive atrophy of the gastrocnemius muscle without any change in the total DNA content of the whole muscle. Histamine content of the muscle decreased; glutamic and acid content increased and histidine level remained unaltered on denervation. Histaminase activity localized in the muscle decreased; glutamic acid content increased and histidine level remained unaltered on denervation. Histaminase activity localized in the muscle mitochondria increased on denervation. The histidine-degrading enzymes, histidine ammonia lyase, urocanate hydratase and imidazol-5yl lactate dehydrogenase, are localized in the sarcoplasm of the muscle and their activities are not altered on denervation. The histidine decarboxylase activity localized in the mitochondria is not altered on denervation. The reduction in the histamine content of the atrophied muscle may be due to increased mitochondrial histaminase activity but not due to increased decarboxylation of histidine. The loss of 'trophic influence' due to denervation may be manifested in the impairment of mitochondrial histaminase activity.

Acetylcholine↗

PubMed1978

Substances moved by axonal transport and released by nerve stimulation have an innervation-like effect on muscle.

Substances which have an innervation-like effect on the cholinesterase activity of organ-cultured rat extensor digitorum longus muscles are moved in nerve by axonal transport, are released from nerve by stimulation, and are present in innervated muscle but apparently absent from denervated muscle. Substances which increase the acetylcholine sensitivity of cultured muscles behave similarly.

Acetylcholine↗

PubMed1971

Ionic permeability changes induced by some cholinergic agonists on normal and denervated frog muscles.

1. The reversal potential of responses to iontophoretic application of acetylcholine, carbamylcholine and decamethonium has been measured at junctional and extrajunctional sensitive areas of the normal sartorius muscle of the frog.2. The values obtained with any one of the drugs at junctional spots were similar to those previously reported for the end-plate potential (about - 16 mV).3. On the other hand, at extrajunctional areas the reversal potential of responses to these agonists was found to have a mean value of - 42 mV. The muscle-tendon responses to acetylcholine reversed at - 51 mV.4. The response to decamethonium was used to determine the nature of the ionic permeability changes mediated by the extrajunctional receptors. The results obtained show that extrajunctional responses are due to an increase in conductance to Na(+) and K(+) but not to Cl(-).5. The reversal potential of acetylcholine responses in denervated frog sartorius was - 42 mV both at former junctional spots and at non-neural sensitized regions.6. These different values of the reversal potential can be due either to effects of receptor density on the ionic selectivity of the membrane or to regional variations in the extracellular ionic accumulation during the transmitter action.

Acetylcholine↗

PubMed1991

Calcium action potentials in innervated and denervated rat muscle fibres.

We studied the generation of calcium action potentials (Ca APs) in innervated and denervated fibres of the extensor digitorum longus of the rat in a tetraethylammonium (TEA) sulphate solution plus 3-4 diaminopiridine (3-4 DAP). The main results are the following: (1) more than 90% of the innervated fibres were capable of developing well-sustained Ca APs that were blocked by Cd or nifedipine; (2) the incidence of Ca APs obtained from the denervated fibres was substantially lower than in the control preparations; (3) no relation was found between the appearance of Ca APs in the denervated fibres and the resting membrane potential (Vm), stimulus duration (500-2000 ms) or holding potential (-80, -100 mV); (4) The percentage of denervated fibres that exhibited Ca APs was increased significantly with the following procedures. First, by raising the external Ca concentration to 14 mM; second, by depleting the intracellular K concentration by overnight exposure of the muscles to a free K-Cs solution; (c) and third, by incubating the muscles in 500 nM apamin, a venom that inhibits the K conductance activated by Ca. Several factors may be involved in the lower incidence of Ca APs obtained in denervated fibres: (1) a diminished Ca current due to a reduction in the driving force as a result of an increment in the intracellular Ca concentration; (2) a persistence of a shunting K conductance that is not inhibited by TEA and 3-4 DAP; (3) a shift in the voltage dependence of the activation and inactivation parameters of the Ca current or the appearance of a new type of Ca channel with a different kinetics.

4-Aminopyridine↗

PubMed1985

Neural cell adhesion molecule (N-CAM) accumulates in denervated and paralyzed skeletal muscles.

We have used immunofluorescence and immunoblotting methods to study the amount and distribution of the neural cell adhesion molecule (N-CAM) in rat skeletal muscle; this molecule is thought to mediate adhesion of neurons to cultured myotubes. N-CAM is present on the surface of embryonic myotubes, but it is lost as development proceeds and is nearly absent from adult muscle. However, denervation of adult muscle results in the reappearance of N-CAM. In denervated muscle, N-CAM is associated both with muscle fibers and with cells in interstitial spaces between fibers. The N-CAM in interstitial spaces is concentrated near denervated endplates, which are known to be preferential sites for reinnervation. Paralysis of innervated muscle, known to mimic denervation in many respects, also induces the accumulation of N-CAM. Axons that regenerate to reinnervate muscle bear N-CAM on their terminals, and reinnervation results in the disappearance of N-CAM from muscle. Denervation induces accumulation of N-CAM in mouse and chicken, as well as in rat muscles. Thus, the expression of N-CAM in muscle is regulated by the muscle's state of innervation. In that N-CAM-rich muscles (embryonic, denervated, and paralyzed) are known to be competent to accept synapses, while N-CAM-poor muscles (normal adult and reinnervated) are refractory to hyperinnervation, N-CAM might, in turn, participate in regulating muscle's susceptibility to innervation.

Animals↗

PubMed1995

Expression and distribution of sodium channels in short- and long-term denervated rodent skeletal muscles.

1. Loose-patch voltage-clamp recordings were made from rat and mouse skeletal muscle fibres denervated for up to 6 weeks. Innervated muscles possessed a Na+ current density of 107 +/- 3.3 mA cm-2 in endplate membrane, and 6.3 +/- 0.6 mA cm-2 in extrajunctional membrane. This high concentration of Na+ channels at the endplate was gradually reduced following denervation. After 6 weeks of denervation, the endplate Na+ channel concentration was reduced by 40-50%, and the density of Na+ channels in extrajunctional membrane was increased by about 30%. 2. The tetrodotoxin (TTX)-resistant form of the Na+ channel appeared after 3 days of denervation and comprised approximately 43% of the endplate Na+ channels 5-6 days after denervation. Subsequently, TTX-resistant Na+ channels were reduced in density to approximately 25% of the postjunctional Na+ channels and remained at this level up to 6 weeks after denervation. 3. RNase protection analysis showed that mRNA encoding the TTX-resistant Na+ channel was virtually absent in innervated muscle, rose > 50-fold after 3 days of denervation, then decreased by 95% 6 weeks after denervation. The density of TTX-resistant Na+ channels correlated qualitatively with changes in mRNA levels. 4. These results suggest that the density of Na+ channels at neuromuscular junctions is maintained by two mechanisms, one influenced by the nerve terminal and the other independent of innervation.

Animals↗

PubMed1994

Parvalbumin immunohistochemistry in denervated skeletal muscle.

Parvalbumin is a calcium-binding protein which, in muscle, is mainly found in type 2B fibres, whereas type 1 fibres lack parvalbumin immunoreactivity. Previous studies have shown that this pattern is highly dependent upon motor neuron innervation and is modified in denervated, cross-reinnervated or chronic low-frequency stimulated muscles. In the present study, we have examined the modifications of parvalbumin immunocytochemistry in the anterior tibialis muscle of the rat at different intervals following section of the sciatic nerve. During the first 2 weeks after denervation, no changes in parvalbumin immunoreactivity were seen, although a global reduction of fibre diameter was observed. Three weeks after denervation, small angulated, strongly parvalbumin-immunoreactive fibres appeared. From the second month onwards, the pattern of parvalbumin immunohistochemistry was characterized by areas composed of small, strongly immunoreactive fibres separated by less atrophic areas displaying a normal chequerboard distribution of parvalbumin immunoreactivity. The increase of parvalbumin-immunoreactivity in denervated and reinnervated muscle, as seen in our study, indicates that important changes in parvalbumin distribution occurs in muscle fibres after denervation. These changes are probably produced in an attempt to bind the free cytosolic calcium which accumulates in denervated fibres, and further reinforces the role of parvalbumin in calcium homeostasis during denervation and reinnervation.

Adenosine Triphosphatases↗

PubMedDate not supplied

Beta-adrenoreceptors in denervated skeletal muscles of the cat.

1. A number of sympathomimetic amines have been compared with (-)-isoprenaline for their ability to produce contractions in chronically denervated soleus and tibialis anterior muscles and to increase heart rate in bilaterally vagotomized anaesthetized cats. 2. The mean dose-ratios ((-)-isoprenaline equals 1) for adrenaline, orciprenaline and salbutamol were 2.9, 29.7 and 11.5 respectively in the tibialis anterior muscle and 3.5, 24.1 and 14.3 in the soleus muscle. The dose-ratios for noradrenaline were 21.1 in the tibialis anterior and 57.6 in the soleus muscle. 3. The dose-ratios ((-)-isoprenaline equals 1) obtained for the positive chronotropic effects of the drugs in bilaterally vagotomized cats were 14.5, 14.6, 29.6 and 24.2 for adrenaline, noradrenaline, orciprenaline and salbutamol respectively. 4. The beta2-receptor antagonists butoxamine and H35/25 antagonized the vasodepressor and skeletal muscle responses to (-)-isoprenaline to a greater extent than the cardiac responses, whereas the beta1-receptor selective antagonists practolol and H93/26 antagonized cardiac to a greater extent than vascular and skeletal muscle responses. 5. The results obtained suggest that the contractions of denervated skeletal muscle to sympathomimetic drugs result from stimulation of beta2-adrenoreceptors.

Adrenergic beta-Agonists↗

PubMed1995

Expression of alpha-cardiac myosin heavy chain in normal and denervated rat muscle spindles.

Whether any fibers in rat hindlimb muscles express alpha-cardiac myosin heavy chain (MHC) is uncertain. Expression of alpha-cardiac MHC mRNA and the polypeptide for which it codes were examined in control and denervated rat muscle spindles using in situ hybridization (ISH) and immunocytochemistry (ICC). Both nuclear bag2 and bag1 intrafusal fibers in the extensor digitorum longus (EDL) muscles expressed alpha-cardiac MHC and its precursor mRNA. Furthermore, denervation of the hindlimb down-regulated alpha-cardiac MHC mRNA expression in rat nuclear bag intrafusal fibers, even though they continued to display a strong affinity for anti-alpha-cardiac MHC monoclonal antibody. These data show that (1) intrafusal fibers express the alpha-cardiac MHC gene; (2) innervation regulates alpha-cardiac MHC gene expression at a pre-translational level; and (3) ISH is more sensitive than ICC to changes in alpha-cardiac MHC gene expression in adult rat muscle spindles.

Animals↗

PubMed1976

Junctional and extrajunctional acetylcholine receptors in normal and denervated frog muscle fibres. Noise analysis experiments with different agonists.

Ionic channel properties of acetylcholine receptors located in, in the vicinity of, or far away from a frog neuromuscular junction were investigated by noise analysis of drug induced current fluctuations. For drugs applied to the junction, in certain cases two Lorentzian curves were necessary to describe the data. It is postulated that the reason for this observation is that a contribution from perijunctional receptors was being observed. The conductance of a single channel in the junction was independent of the nature of the agonist and had an average value of 17.9 pS (temperature range 8-25 degrees C, solution buffered with Tris). After denervation for 21 days the conductance gamma was 7.5 pS at extrajunctional locations. In the close neighbourhood of the junction (peri-junctional receptors) values were found between 4 and 19 pS. The mean value of the open channel life-time tau in the endplate exposed to acetylcholine was 2.4 ms at 8-11 degrees C. This value was 0.90 ms with carbachol, 0.50 ms with succinylcholine, 0.28 ms with decamethonium and 0.45 ms with nicotine. The receptors outside the endplate exhibited tau-values which at a given temperature were 2-3 times larger than those at the endplate. Raising the temperature to 23 degrees C reduced all tau-values by factors of 2-3. It is concluded that at least two types of ACh-receptors with different properties exist in the muscle membrane, possibly produced by ACh-receptive units in different states of aggregation.

Acetylcholine↗

PubMed1983

Selectivity of sodium channels in denervated tonic muscle fibre membrane of the frog.

Ionic selectivity of sodium channels was examined under voltage clamp conditions in normal and denervated twitch fibres and denervated tonic fibres isolated from m. ileofibularis of the frog (R. temporaria). Membrane currents were recorded by means of the Hille-Campbell vaseline-gap voltage clamp method from muscle fibre segments exposed to a potassium-free artificial internal solution. Permeability ratio (PS/PNa) were determined from changes in the reversal potential after replacing all Na ions in the solution bathing the voltage clamped external membrane area with sodium substituting ions (S). The permeability sequence was: Na+ greater than Li+ greater than NH4+ greater than K+. No inward currents were observed for Ca2+. The permeability ratios were as follows. Denervated tonic fibres: 1:0.88:0.23:0.012; control twitch fibres: 1:0.94:0.22:0.076; denervated twitch fibres: 1:0.91:0.14:0.082. The permeability to Li+ ions deviates from independence to a greater extent in tonic than in phasic fibres. Our results are consistent with the Hille model of sodium channel selectivity, and they support the hypothesis that sodium channels formed in denervated tonic muscle fibres of the frog are of the same genetic origin as Na channels expressed under physiological conditions.

Animals↗

PubMed1987

Fetal-type creatine kinase in rat fast and slow muscles during denervation and reinnervation.

Creatine kinase (CK) has three forms of isozymes; CK-BB, CK-MB, and CK-MM. In adult rats they show a specific tissue distribution: the BB form in the brain, the MB form in the heart, and the MM form in skeletal muscle. In embryonic skeletal muscles only the BB and MB forms are found. Adult slow-twitch muscles contain more fetal type creatine kinase (CK-B) than do fast-twitch muscles. In the present experiment the effect of denervation and reinnervation on the CK-B concentration was investigated in rat fast (extensor digitorum longus)- and slow (soleus)-twitch muscles by a highly sensitive immunoassay. Denervation of these muscles produced a progressive increase in CK-B concentration in both muscles. When the sciatic nerve was cut and immediately sutured, the CK-B concentration in both muscles showed a gradual reduction after an initial increase. By the 34th postoperative week the CK-B concentration in the soleus was about one-half that of the contralateral control, whereas that in the extensor digitorum longus was nearly normal. After cross union of the nerves innervating the muscles, the CK-B concentration in the soleus was reduced at 35 weeks to about one-half normal, but that in the extensor digitorum longus was always higher than the control value. After self-reunion of the nerves, the CK-B concentration at the 20th week was approximately normal in the extensor digitorum longus and significantly increased in the soleus. We suggest that the motoneurons normally innervating the extensor digitorum longus have a greater capability in suppressing the production of CK-B than do the soleus motoneurons.

Animals↗