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PubMed1995

Ca2+ release channels in rat denervated skeletal muscles.

Sarcoplasmic reticulum (SR) Ca2+ release channel-ryanodine receptors (RYR1) from rat fast-twitch skeletal muscle were studied by incorporating heavy sarcoplasmic reticulum membranes into a lipid bilayer. Channels from normal and denervated muscles had the same conductance as that reported for rabbits (about 500 pS) in 250:250 mM cis:trans caesium methanesulphonate. Caffeine (0.1 mM) induced a larger increase in the open probability (Po) in denervated than in normal channels. The caffeine effect was caused by changes in mean open and burst time distributions. Longer opening and burst events were detected in the presence of caffeine. High caffeine concentrations (4 mM) gave similar results in channels from normal and denervated muscles. In denervated muscle, unlike intact muscle, the Ca2+ release channel was not activated at millimolar Ca2+ concentrations; this is similar to the cardiac isoform of the channel. Maximal channel activation was shifted to higher Ca2+ concentrations (pCa 4) and the channel remained activated at millimolar Ca2+ concentrations. The main effect of millimolar Ca2+ concentrations upon Ca2+ release channels from denervated muscles was an increase in the mean open time, with a concomitant increment of the mean burst duration. Alterations in channel gating properties in calcium and caffeine account for changes in the mechanical response after skeletal muscle denervation.

Animals↗
PubMed1998

Succinylcholine-induced fasciculations in denervated rat muscles as measured using 31P-NMR spectroscopy: the effect of pretreatment with dantrolene or vecuronium.

BACKGROUND: We have previously demonstrated by 31P nuclear magnetic resonance (NMR) that succinylcholine (SCh) induces metabolic changes in denervated muscle. To specify those changes, we attempted to inhibit them using two different kinds of drugs, dantrolene and vecuronium. METHODS: Three weeks after unilateral sciatic nerve section, 75 male Wistar rats were randomly assigned to one of the following 5 groups: (1) non-pretreated normal muscle group; (2) non-pretreated denervated muscle group; (3) denervated muscle group pretreated with a low dose of vecuronium (0.02 mg.kg(-1)); (4) denervated muscle group pretreated with a high dose of vecuronium (0.2 mg.kg(-1)); (5) denervated muscle group pretreated with dantrolene (2 mg.kg(-1)). The change of the inorganic phosphate/phosphocreatine (Pi/PCr) ratio of each muscle was measured by 31P-NMR before and after SCh (1 mg.kg(-1)) administration and the corresponding peak amplitude of the electromyograms (EMG) was determined. RESULTS: The high dose of vecuronium totally inhibited SCh-induced fasciculation on EMG (100%-->2%). In this group, though the Pi/PCr ratio significantly increased 10 min after SCh, the peak after 5 min disappeared. The inhibition with dantrolene was about the same order of magnitude as with the low dose of vecuronium (35%:21%). However, the increase in the Pi/PCr only lasted about 10 min, in contrast to the other drugs. CONCLUSION: Our findings indicate that the Pi/PCr increases 5 and 10 min after SCh, respectively, as a result of two different processes. The first peak is caused by an excessive energy consumption in response to excessive muscle contraction. This in turn triggers the second peak, caused by breakdown of glycogen, initiated by an increased Ca2+ concentration.

Animals↗
PubMed1983

Regulation of acetylcholine receptor density in membranes of denervated mouse muscles.

The ACh sensitivity of denervated muscles of rats stimulated in situ or in culture for several days decreases to low levels characteristic of normal muscles. Possible causes are the electrical activity, depolarization per se, contraction and intracellular [Ca2+] changes. To test the last three hypotheses, isolated bundles of denervated mouse soleus muscles were placed in narrow chambers (internal diameter 2 mm) which were periodically perfused with standard mixtures of Minimal Essential Medium (MEM) and calf serum. At 10-15-min intervals the chambers were filled for 15-43 s with a Ringer solution containing 55 microM ACh or 8-10 mM caffeine or with a solution containing 200 mM KCl. After 3-7 days the muscles were tested for ACh sensitivity by comparing the force developed in response to 110 microM ACh and that to 400 mM K methanesulphonate. In addition, the ACh receptor density was measured with 125I alpha-bungarotoxin. The results showed no difference in ACh sensitivity or receptor density between treated and untreated muscles. Small differences produced by caffeine were probably caused by muscle damage. Ca influx, but not efflux, was strongly elevated during exposure to ACh. It was concluded that neither depolarization per se nor contractile activity or the associated calcium movements have any effect on ACh receptors.

Acetylcholine↗
PubMed2001

Small-conductance calcium-activated potassium currents in mouse hyperexcitable denervated skeletal muscle.

1. Hyperexcitability in denervated skeletal muscle is associated with the expression of SK3, a small-conductance Ca2+-activated K+ channel (SK channel). SK currents were examined in dissociated fibres from flexor digitorum brevis (FDB) muscle using the whole-cell patch clamp configuration. 2. Depolarization activated a K+-selective, apamin-sensitive and iberiotoxin-insensitive current, detected as a tail current upon repolarization, in fibres from denervated but not innervated muscle. Dialysis of the fibres with 20 mM EGTA in the patch pipette solution eliminated the tail current, consistent with this current reflecting Ca2+-activated SK channels expressed only in denervated muscle. 3. Activation of SK tail currents depended on the duration of the depolarizing pulse, consistent with a rise in intracellular Ca2+ due to release from the sarcoplasmic reticulum (SR) and influx through voltage-gated Ca2+ channels. 4. The envelope of SK tail currents was diminished by 10 microM ryanodine for all pulse durations, whereas 2 mM cobalt reduced the SK tail current for pulses greater than 80 ms, demonstrating that Ca2+ release from the SR during short pulses primarily activated SK channels. 5. In current clamp mode with the resting membrane potential set at -70 mV, denervation decreased the action potential threshold by approximately 8 mV. Application of apamin increased the action potential threshold in denervated fibres to that measured in innervated fibres, suggesting that SK channel activity modulates the apparent action potential threshold. 6. These results are consistent with a model in which SK channel activity in the T-tubules of denervated skeletal muscle causes a local increase in K+ concentration that results in hyperexcitability.

Action Potentials↗
PubMed1993

Impact of electropolarity treatment on contractile kinetics of gastrocnemius muscle of toad (Bufo melanostictus) during denervation.

The twitch properties of denervated muscle were found to be altered contributing to the derangement in the contractile machinery of the denervated muscle. Cathodal polarity treatment minimized the fluctuations and tried to restore denervated muscle back to normalcy to some extent. The anodal polarity treatment was not able to produce favourable effect. The differential effect of polarity treatment on the contractile kinetics of the denervated muscle is discussed.

Animals↗
PubMed1974

Dynamic properties of denervated fast and slow twitch muscle of the cat.

1. Isometric and isotonic contractions were recorded from cat flexor digitorum longus (FDL) and soleus muscles denervated for 28-35 days and compared with the responses of contralateral control muscles.2. Isometric twitch time to peak was longer in the denervated muscles than in the controls. Tetanic tension was reduced whether expressed as absolute units, per gram or per unit area. Absolute twitch tensions were reduced less below the control values with a consequent increase of twitch-tetanus ratio. These results agree with earlier reports; but, in contrast, there was no reduction in the relative rate of rise of tension in the isometric tetanus of denervated compared with control FDL. The maximum rate of rise of tension in the twitch was only a little less than that in the fully fused tetanus in the denervated muscles.3. Whole muscle shortening velocity was reduced in the denervated soleus with no change in the number of sarcomeres per fibre. There was a smaller change in FDL muscles. This was the result of an increase in the number of sarcomeres per fibre which followed denervation in FDL, compensating for a decreased velocity of shortening of the sarcomere. Maximum sarcomere shortening velocity was reduced by 30% relative to control values in both muscles.4. Measurements were made of the isometric contractions resulting from pairs of stimuli in an attempt to assess the period of maximal activation in the twitch. Maximum summation occurred when the second stimulus was given close to the time of the peak of the twitch in the denervated muscles.

Animals↗
PubMed1986

Electrophysiologic denervation changes of human muscle fibers in motoneuron diseases.

Muscle denervation and reinnervation by sprouting of the surviving motoneurons characterizes motoneuron diseases (MND). In mammalian muscles, experimental denervation induces the appearance of extrajunctional acetylcholine (ACh) receptors and tetrodotoxin (TTX) resistant action potentials (AP). These denervation changes have been investigated in muscle biopsies from 10 MND patients and in 2 traumatically denervated normal human muscles. Extrajunctional ACh sensitivity was present in 113 of the 140 (73%) fibers from MND muscles studied. In 50 of 84 (70%) ACh-sensitive fibers, no TTX-resistant AP were present. The remaining fibers (30%) showed small regenerative responses. In contrast, all the traumatically denervated muscle fibers showed extrajunctional ACh sensitivity and TTX-resistant AP. Histochemical analysis of the biopsies showed no direct correlation between the frequency of ACh-sensitive fibers and that of the atrophic or normal-appearing fibers. The absence of TTX-resistant AP in ACh-sensitive fibers and its lack of correlation with the histochemical criteria of denervation suggest the presence of a state of innervation in MMD, where the motoneuron is not able to maintain its fully trophic influence on the muscle fiber membrane.

Acetylcholine↗
PubMed1997

Changes in myosin expression in denervated laryngeal muscle.

The effects of chronic denervation on the myosin heavy chain (MyHC) content and muscle fiber type composition of rat laryngeal muscles are described. The posterior cricoarytenoid (PCA) and thyroarytenoid (TA) muscles were removed 3 weeks, 3 months, and 6 months after recurrent laryngeal nerve sectioning. Myofibrillar adenosine triphosphatase staining of cryostat sections was performed, and fiber type percentages were determined. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to separate MyHC isoforms, and densitometry was subsequently used for quantitative analysis. Unoperated animals served as controls. In the PCA muscle, denervation resulted in a progressive reduction in type I MyHC (the slow-contracting isoform) to an almost complete loss at 6 months, with a concomitant increase in type II MyHCs (fast-contracting isoforms, excluding type IIL). Type IIL MyHC (laryngeal-specific isoform) remained relatively constant up to 6 months after denervation. The myosin expression in the TA muscle, which contained only type II MyHCs, remained relatively constant with denervation. Changes in fiber type composition of the muscles described from tissue staining correlated with MyHC content. These findings in laryngeal muscle confirm the dependence of type I MyHC expression upon neural input, as has been found previously in limb skeletal muscles. Since the expression of all MyHCs except the IIL was modified after denervation in the PCA muscle, it is possible that the IIL isoform is maintained by factors that differ from those in the other skeletal myosins.

Animals↗
PubMed1999

Late-preconditioning protection is evident in the microcirculation of denervated skeletal muscle.

We investigated whether ischemic preconditioning induces microvascular protection in skeletal muscle at the late phase (after 24 hours) when the same muscles are subjected to prolonged warm global ischemia. The cremaster muscle of the male Sprague-Dawley rat underwent vascular isolation and was subjected to 4 hours of ischemia and 60 minutes of reperfusion. Early preconditioning consisted of 45 minutes of ischemia followed by 15 minutes of reperfusion before prolonged ischemia/reperfusion; late preconditioning also consisted of 45 minutes of ischemia but was done 24 hours (24-hour period of reperfusion) before the prolonged ischemia/reperfusion. Arteriole diameters and capillary perfusion were measured with use of intravital microscopy. Four groups were compared: rats that underwent early preconditioning, their controls, rats that underwent late preconditioning, and their controls. Early and late preconditioning significantly attenuated vasospasm and capillary no-reflow compared with the controls for each. Average arteriole diameter was significantly larger in the rats that underwent late preconditioning than in any other rats; it was also significantly larger in the controls for late preconditioning than in those for early preconditioning. We introduce a model of the rat cremaster muscle that has been isolated from its vascular supply as a useful preparation to study the effects of late preconditioning on microcirculation in skeletal muscle. Late preconditioning provided better microvascular protection than did early preconditioning. The mechanism for this preconditioning protection is being investigated because it should provide a means for therapeutic intervention.

Animals↗
PubMed1982

Membrane specialisation of the extra-junctional sarcolemma of normal and denervated insect muscle.

Corrugated areas of sarcolemma were observed in serial transverse sections. Complexes of intramembranous particles that appear after freeze-fracture replication were concluded to represent the same specialisation. These specialisations, up to 14 microns x 2 microns, are orientated parallel to the long axis of the muscle. Intramembranous particles are concentrated along the peaks of the corrugations, and are associated with the P-face. Corresponding pits are found in the E-face. Fourteen and thirty days after sectioning the excitatory motor-nerve supply to the muscle, corrugated areas 0.5-1 microns x 0.5-1 microns are found. Occurring singly or in groups, their orientation with respect to the long axis of the muscle is more variable than those of control muscles. Thin sections reveal no complementary areas on adjacent fibres or intracellular submembrane attachments or specialisations. A structural role is therefore unlikely. Mitochondria are frequently found in close association with these specialisations. Their possible role as receptors or transmembrane transport systems is discussed.

Animals↗
PubMed1981

Increased phosphorylation in vitro of a cytosolic polypeptide resolved from denervated skeletal muscle.

The in vitro phosphorylation of a 40,400-dalton, cytosolic polypeptide from the soleus muscle of the rat is increased twofold within 24 hr after cutting the motor nerve fibers to this muscle. This involves an ATP-phosphotransferase reaction which we have reported to be inhibited by a specific cyclic AMP-dependent protein kinase inhibitor. The phosphorylated polypeptide does not electrophoretically comigrate on SDS-polyacrylamide gels with the 38,000-dalton catalytic subunit of cyclic AMP-dependent protein kinase which is known to undergo a site-specific autophosphorylation in skeletal muscle.

Animals↗