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PubMed1980

Effects of denervation and botulinum toxin on muscle sensitivity to acetylcholine and acceptance of foreign innervation in the frog.

1. The effects of denervation and local paralysis produced by botulinum toxin (type D) on the sensitivity of skeletal muscle to ACh and its ability to accept innervation by an implanted foreign nerve were investigated in the frog. 2. Denervated muscles developed supersensitivity to ACh within 2 weeks and became extensively innervated by an implanted foreign nerve after about 4 weeks. 3. Chronic electrical stimulation of denervated muscles (50 Hz for 1 sec every 60 sec) did not prevent the development of supersensitivity. 4. Muscles paralysed by botulinum toxin did not usually develop supersensitivity to ACh until after 2-3 months and the extra-junctional sensitivity of individual fibres was generally less than after denervation. Significant innervation of the paralysed muscles by an implanted foreign nerve did not occur until after 2-3 months. 5. The results suggest that in the frog nerves are able to control muscle sensitivity to ACh and to prevent innervation by foreign nerves by some mechanism other than muscle activity. Prolonged inactivity seems to result in some development of extra-junctional sensitivity and acceptance of foreign innervation.

Acetylcholine↗
PubMed1981

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↗
PubMed1984

Turnover of acetylcholinesterase in innervated and denervated rat diaphragm.

The acetylcholinesterase (AChE) in rat diaphragms was labelled by intravenous injection of echothiophate in order to evaluate the turnover of AChE in innervated and denervated muscle in vivo. Echothiophate diethylphosphorylates AChE thereby inactivating it. Labelled (diethylphosphorylated) enzyme is rapidly and quantitatively reactivated with 1-methyl-2-hydroxyiminomethylpyridinium (2-PAM), so labelled (diethylphosphorylated) AChE was conveniently measured as 2-PAM-reactivatable AChE activity. In homogenates in vitro, label is lost spontaneously (diethylphosphorylated AChE spontaneously reactivates) with a half-time of 27 h. In innervated diaphragm, labelled non-end-plate AChE is lost with a half-time of 13 h. When correction is made for the spontaneous loss of label on the basis of in vitro measurements, this data indicates that non-end-plate AChE turns over with a half-time of about 26 h. In innervated diaphragm, labelled end-plate-specific AChE is lost more slowly than non-end-plate AChE and at a rate essentially identical to the rate of spontaneous loss of label in vitro. The rate of loss of labelled non-end-plate AChE is essentially identical in 18 h denervated and in paired innervated diaphragms. The rate of loss of labelled end-plate-specific AChE is significantly faster in 18 h denervated diaphragms than in paired innervated diaphragms. On the basis of these observations, hypotheses concerning the mechanisms of the denervation-induced decreases in non-end-plate and end-plate-specific AChE are formulated and discussed.

Acetylcholinesterase↗
PubMed1988

Post-natal disappearance of transient calcium channels in mouse skeletal muscle: effects of denervation and culture.

1. The whole-cell voltage clamp technique was used to record Ba2+ currents in voltage-sensitive Ca2+ channels in mouse flexor digitorum brevis muscles developing in situ from day 1 to 30 after birth. Effects of denervation and tissue culture on the Ca2+ channel currents were also studied. 2. The muscle fibres in newborn mice showed two distinct types of Ca2+ channel currents, a low-threshold transient current and a high-threshold sustained current. 3. The specific amplitude of the transient current was 2.7 +/- 1.7 (S.D.) A/F in response to -30 mV test pulses in medium containing 30 mM-Ba2+ on day 1 after birth. The transient current decreased progressively in the post-natal days and became undetectable by day 17. In contrast, the specific amplitude of the sustained current in response to +20 mV test pulses increased 4-fold from 6.9 A/F on day 1 to 27.7 A/F on day 30. 4. The disappearance of the transient current could not be accounted for by either shifts in voltage dependence of activation and inactivation or changes in activation and inactivation times of the two types of current during development. 5. Denervating muscle fibres on day 8 after birth did not prevent the disappearance of the transient current. Denervating them on day 17 did not allow reappearance of the transient current. However, the increase of the sustained current was suppressed by the denervation either on day 8 or day 17. 6. In muscle fibres isolated on day 8 after birth and cultured thereafter, the transient current did not disappear until day 19 in culture (27 days after birth), while the sustained current was maintained at the level on day 8. 7. In muscle fibres isolated on day 17, when the transient current had become undetectable, and cultured thereafter, the transient current did not reappear until day 15 in culture (32 days after birth), while the sustained current was maintained at a level similar to that on day 17. 8. We conclude that innervation has little influence on the developmental disappearance of the transient Ca2+ channel current in mouse muscle fibres, and suggest that some influencing factors from surroundings other than the nerve may be required for the disappearance of the functional transient channels.

Action Potentials↗
PubMed1992

The role of acetylcholinesterase in denervation supersensitivity in the frog cardiac ganglion.

1. The sensitivity of normal and denervated cardiac ganglion cells to the cholinergic agonists acetylcholine and carbamylcholine (carbachol) were compared in the frog, Rana pipiens. Acetylcholine and carbachol bind to the same acetylcholine receptors, but, unlike acetylcholine, carbachol is resistant to hydrolysis by acetylcholinesterase. 2. Sensitivity was assessed by the peak depolarization elicited in response to a sustained pulse of ligand emitted from a pipette positioned 10 microns from the ganglion cell surface. This technique allows the sensitivity of the entire cell to be recorded with a single measurement. 3. The acetylcholine sensitivity of normal cardiac ganglion cells was increased by inhibiting extracellular acetylcholinesterase with echothiophate. 4. Denervation increased the sensitivity of cardiac ganglion cells to acetylcholine but not to carbachol. 5. Following the inhibition of extracellular acetylcholinesterase with echothiophate, sensitivity to acetylcholine was similar in normal and in denervated ganglion cells. 6. The increased sensitivity to acetylcholine of cardiac ganglion cells following denervation is caused by a reduction in the hydrolysis of the transmitter by acetylcholinesterase rather than by changes in the number and/or properties of acetylcholine receptors.

Acetylcholine↗
PubMed1992

The importance of competition between motoneurones in developing rat muscle; effects of partial denervation at birth.

1. The number of motor units in developing fourth deep lumbrical muscles was reduced by unilateral partial denervation of the muscle at birth, by cutting the lateral plantar nerve. A minority of the motor axons arrive via the sural nerve, and were thus not cut. Those muscles that contained one motor unit (one-unit muscles) after partial denervation developed in the absence of competition between motoneurones. Muscles with two motor units had little competition. A few four-unit muscles were studied for comparison. 2. Isometric twitch and tetanic tensions of single motor units were recorded in vitro at 60 days of age in response to stimulation of the sural nerve. On average, units in partially denervated muscles generated more tension than normal units. The isometric tension characteristics of the units in the one-unit and two-unit muscles were different from the normal units (e.g. slower contracting and more fatiguable). The units of four-unit muscles had properties similar to those of normal muscles. 3. Fibres of an individual unit were identified by glycogen depletion and S (slow) fibres were identified in cross-section that bound a polyclonal antibody to slow type I myosin. Those fibres that did not bind the antibody were designated F fibres. The units of one-unit muscles had the same total number of fibres and fibre type composition (both S and F fibres in the same unit) as estimated from previous work to exist at birth. The units of two-unit muscles contained the same total number of fibres, but apparently fewer S fibres, though this may have been as a result of incomplete glycogen depletion of some fibres. 4. It is concluded that competition between motoneurone terminals is necessary for the withdrawal of mismatched connections on muscle fibres present at birth; or, alternatively, that such withdrawal cannot take place if it would result in denervation of the muscle fibre.

Animals↗
PubMed1994

Effect of carotid denervation on plasma vasopressin levels during acute hypoxia in the late-gestation sheep fetus.

1. We measured plasma concentrations of arginine vasopressin (AVP), arterial, venous and amniotic pressures, and carotid and femoral blood flows in fifteen chronically instrumented fetal sheep at 119-125 days of gestation. In eight of the fetuses the carotid sinus nerves were cut (denervated fetuses); the other seven remained intact and served as controls (intact fetuses). 2. In the intact fetuses during hypoxia there was an increase in plasma [AVP] and in perfusion (arterial-venous) pressure, a transient bradycardia, and an increase in carotid and a decrease in femoral blood flow. Whilst femoral vascular resistance (perfusion pressure/femoral blood flow) increased, there were no changes in carotid vascular resistance during hypoxia. 3. In the denervated fetuses no significant bradycardia, fall in femoral blood flow or increase in femoral vascular resistance was present soon after the onset of hypoxia but plasma AVP increased to similar concentrations to those observed in intact fetuses during hypoxia. 4. We conclude that carotid denervation does not affect plasma [AVP] during hypoxia in fetal sheep. This suggests that (1) AVP release during hypoxia is not mediated by a carotid chemoreflex and (2) AVP does not play an important role in these initial fetal cardiovascular responses. Furthermore, we previously reported that intact fetuses survive acute hypoxia better than denervated fetuses following phentolamine treatment, and we believe this to be due to the action of a non-alpha-adrenergic vasoconstrictor released in part via a carotid chemoreflex. The present results suggest that this vasoconstrictor is not AVP.

Acute Disease↗
PubMed1995

The effect of partial denervation of developing rats fast muscles on their motor unit properties.

1. The effects of partial denervation on motor units of the fast twitch extensor digitorum longus (EDL) muscle of the rat were studied. 2. Partial denervation was performed by surgically removing 2-4 mm of the L4 ventral ramus in 3- and 18-day-old Wistar rats. Two to three months later, EMG activity, contractile properties and muscle fibre types were analysed. 3. After partial denervation the EDL muscle became significantly more active, particularly in the animals operated on at 3 days. The amount of activity during periods of rest was up to 4 times that of the control EDL muscle. 4. The maximum tetanic tension developed by the EDL muscles 2 months after partial denervation at 3 days was only 11 +/- 1.01% (S.E.M., n = 18) of the control. In animals operated on at 18 days this value was larger, i.e. 44 +/- 3.46% (S.E.M., n = 12). The low force output of animals operated on at 3 days was also reflected in the low mean motor unit (MU) force output which was only 69 +/- 5.82% (S.E.M., n = 17) of the contralateral control muscle. In contrast the force generated by MUs of rats operated on at 18 days was larger than that of control muscles, i.e. 151 +/- 13.05% (S.E.M., n = 11). The number of MUs was 6 +/- 0.32 (S.E.M., n = 19) in rats operated on at 3 days and 12 +/- 0.83 (S.E.M., n = 14) in rats operated on at 18 days. 5. The speed of contraction decreased and the resistance to fatigue increased. These changes were greater in animals operated on at 3 days. The proportion of muscle fibres reacting with antibody against slow myosin showed a significant increase, especially in the group of animals operated on at 3 days.

Aging↗
PubMed2004

Protective effect of sensory denervation in inflammatory arthritis (evidence of regulatory neuroimmune pathways in the arthritic joint).

OBJECTIVE: To investigate the direct effect of joint innervation on immune mediated joint inflammation in a patient with psoriatic arthritis (PsA). CASE REPORT: The patient developed arthritis mutilans in all digits of both hands with the exception of the left 4th finger, which had prior sensory denervation following traumatic nerve dissection. Plain radiography, ultrasonography and nerve conduction studies of the hands confirmed the absence of articular disease and sensory innervation in the left 4th digit. METHODS: This relationship between joint innervation and joint inflammation was investigated experimentally by prior surgical sensory denervation of the medial aspect of the knee in six Wistar rats in which carrageenan induced arthritis was subsequently induced. Prior sensory denervation--with preservation of muscle function--prevented the development of inflammatory arthritis in the denervated knee. DISCUSSION: Observations in human and animal inflammatory arthritis suggest that regulatory neuroimmune pathways in the joint are an important mechanism that modulates the clinical expression of inflammatory arthritis.

Animals↗
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↗
PubMedDate not supplied

Differential modulation of carbonic anhydrase (CA III) in slow- and fast-twitch skeletal muscles of rat following denervation and reinnervation.

Carbonic anhydrase III (CA III) is influenced by neuronal factors in skeletal muscles of the rat. CA III protein and its mRNA levels were assessed in slow- and fast-twitch muscles after short-term denervation by ligature of the sciatic nerve and reinnervation following removal of the sheath tightly fixed around the nerve. Significant elevations in the CA III mRNA content of fast-twitch muscles were recorded after denervation, but they were cancelled following spontaneous muscle reinnervation. No such variations were observed in the slow-twitch soleus muscle. CA III specific activity or cytosolic CA III protein content increased in both types of muscles after denervation, while a decrease was solely observed in the soleus after reinnervation. These results suggest that neuronal mediators may be responsible for up and down variations in CA III gene expression and (or) mRNA stability in slow- and fast-twitch muscles exposed to identical stimuli. Variations of the mRNA and the protein probably reflect, in a time-related manner, the well-programmed changes in fiber type of the muscles in the context of the denervation-reinnervation model.

Animals↗
PubMed1980

Lack of effect of chronic renal denervation on altered sodium reabsorption during increased ureteral pressure.

The role of the renal nerves in the altered sodium reabsorption which occurs during increased ureteral pressure was studied using clearance techniques in anaesthetized rats undergoing diuresis induced by isotonic saline infusion. In rats with a sham denervated kidney, an ipsilateral increase in ureteral pressure to 20 cm H2O resulted in a marked and significant decrease in sodium and water excretion, increased fractional sodium reabsorption, and increased urine osmolality with no significant change in glomerular filtration rate. A similar significant ipsilateral increase in tubular reabsorption of sodium occurred in rats with chronically denervated kidneys during increased ureteral pressure. The changes in tubular reabsorption were rapidly reversible after return of ureteral pressure to normal. These experiments indicate that enhanced tubular reabsorption of sodium during an ipsilateral increase in ureteral pressure is not mediated by increased renal nerve activity. During the antinatriuresis of increased ureteral pressure there was a decrease in the fractional reabsorption of sodium from the opposite normal kidney. The role of the renal nerves in this compensatory change in function in the opposite kidney was studied in two further groups of animals. The renal response to a contralateral increase in ureteral pressure was similar in denervated and sham-denervated kidneys. The results indicate that altered renal nerve activity, through ipsilateral or contralateral renorenal reflexes, is not responsible for the changes in tubular reabsorption of sodium which occur during increased ureteral pressure induced by partial ureteral obstruction.

Absorption↗
PubMed1982

Effect of adrenal denervation on the glucose, insulin, and glucagon responses to exercise in sheep.

Previous studies suggest that adrenal catecholamines mediate, in part, the glucose and pancreatic hormonal responses to exercise in sheep. This was examined in sheep whose adrenals were denervated to prevent stress-induced changes in catecholamine secretion. The innervation to the right adrenal gland was severed and the left adrenal was removed. Adrenal denervation was associated with a reduction in exercise-induced hyperglycemia and impairment, as measured by [2-3H]glucose, of the increase in glucose appearance during the first 10 min of exercise and increased metabolic clearance rate of glucose after 20 min of exercise. Insulin concentrations were significantly higher during exercise after adrenal denervation than in the controls. Adrenal denervation did not alter the rise in glucagon due to exercise. These effects are consistent with adrenomedullary hormonal stimulation of hepatic and muscular glycogenolysis, either directly or indirectly through the regulation of insulin secretion during exercise in sheep.

Adrenal Glands↗
PubMed1984

Renal function in rats with innervated and denervated kidneys before and during sodium pentobarbital anesthesia.

We have used a model for measuring renal clearances in the undisturbed rat to assess the role of the renal nerves in the depression of renal function during sodium pentobarbital anesthesia. One group of rats was studied with renal nerves intact and a second group was studied 7-9 days after bilateral renal denervation. Rats were prepared by placement of cannulae an average of 5 days prior to the clearance experiments. Renal function was measured before and after the injection of saline as the control vehicle and 2-3 days later, before, and after the injection of sodium pentobarbital (50 mg/kg) in the same rat. Sodium pentobarbital produced comparable decreases in glomerular filtration rate, para-aminohippuric acid clearance, urine flow rate, and sodium excretion in rats with denervated or innervated kidneys. Injection of saline resulted in no differences in measured variables between the rats with intact or sectioned renal nerves. Sodium pentobarbital caused a drop in arterial pressure in the denervated group but not in the innervated group. In a second series of experiments rats with denervated kidneys were implanted with an inflatable occluder around the aorta. This occluder was inflated to limit the drop in arterial pressure during anesthesia. When the blood pressure to the kidneys was maintained, renal function did not decrease during sodium pentobarbital anesthesia. These experiments suggest that the renal nerves are involved in the decrease in renal function during sodium pentobarbital anesthesia.

Anesthesia↗
PubMed1987

Failure of renal denervation to attenuate hypertension in Dahl NaCl-sensitive rats.

In previous experiments we have demonstrated that the renal nerves play a significant role in all genetic and (or) induced models of hypertension that we have studied. The current experiments extended this research by investigating the contribution of the renal nerves to hypertension in the Dahl NaCl-sensitive rat. This was investigated by assessing the effect of bilateral phenol renal denervation carried out prior to initiation of a high NaCl (8% NaCl) diet. In two separate studies, renal denervation did not affect systolic blood pressure in either Dahl NaCl-sensitive rats or their normotensive counterparts, Dahl NaCl-resistant rats. Further, denervation did not increase absolute urinary sodium excretion, percent urinary sodium excretion, urinary volume output, or food or water intake; nor did it differentially alter creatinine clearance or body weight. Denervation was verified at the termination of each study by a greater than 80% depletion of renal noradrenaline stores. These results indicate that the renal nerves do not provide a major contribution to hypertension in the Dahl NaCl-sensitive rat.

Aging↗
PubMed1991

The effect of partial denervation of tibialis anterior (TA) muscle on the number and sizes of motorneurons in TA motornucleus of normal and dystrophic (C57BL dy2j/dy2j) mice.

The tibialis anterior (TA) muscle in one leg of normal (C57BL) and dystrophic (dy2j) mice was partially denervated by resection of a part of the lateral popliteal nerve. Two months later the muscle was injected with horseradish peroxidase to permit visualization of the motorneurons that survived. Partial denervation in both C57 and dy2j mice resulted in reduction of the number of motorneurons that supplied the muscle to approximately one-half the normal complement. The surviving motorneurons were found to be significantly larger (about 25%) than their contralateral counterparts. This condition persisted up to 18 months and is not considered to be a transient response to the trauma associated with the partial denervation. When the size of the target tissue was also reduced by extirpation of one-half of TA together with partial denervation, motorneuron size was not found to increase. It is suggested that the increase in size is a response to the metabolic demands placed upon the motorneuron by an increase in the size of the motor unit.

Animals↗
PubMed1995

Effect of denervation of the rat superior mesenteric arterial bed on the distribution of resistance among the first four orders of branching.

The rat superior mesenteric artery with its first four orders of branching has been examined as an isolated perfused preparation. Lumen diameter and proportional resistance at each level have been determined and compared under conditions of constant pressure following methoxamine-induced constriction and acetylcholine-induced relaxation in intact and denervated preparations. A comparison was also made between intact preparations under conditions of constant pressure and constant flow. Lumen diameter and segment length were measured by analysis of video-taped images. The distribution of resistance among the first four orders of branching was estimated from the measured values using the Poiseuille formula. Intact first and fourth order branches had a proportionately low resistance, with the remaining resistance being fairly evenly distributed between the second and third order branches. No change in the pattern occurred as a result of either methoxamine or acetylcholine treatment. The pattern of distribution of resistance changed following denervation, so that proportional resistance was high in the first order branches and fell off from the second to fourth order branches. With methoxamine, the first order branches of denervates had a significantly higher resistance than in intact tissues, with the distribution of resistance in the second to fourth order branches following the basal resistance pattern. Reduction in the proportion of resistance with acetylcholine was similar in both intact and denervated preparations. Distribution of resistance did not differ between constant flow and constant pressure intact preparations.

Acetylcholine↗
PubMed2006

Denervation produces different single fiber phenotypes in fast- and slow-twitch hindlimb muscles of the rat.

Using a single, mechanically skinned fiber approach, we tested the hypothesis that denervation (0 to 50 days) of skeletal muscles that do not overlap in fiber type composition [extensor digitorum longus (EDL) and soleus (SOL) muscles of Long-Evans hooded rats] leads to development of different fiber phenotypes. Denervation (50 day) was accompanied by 1) a marked increase in the proportion of hybrid IIB/D fibers (EDL) and I/IIA fibers (SOL) from 30% to >75% in both muscles, and a corresponding decrease in the proportion of pure fibers expressing only one myosin heavy chain (MHC) isoform; 2) complex muscle- and fiber-type specific changes in sarcoplasmic reticulum Ca(2+)-loading level at physiological pCa approximately 7.1, with EDL fibers displaying more consistent changes than SOL fibers; 3) decrease by approximately 50% in specific force of all fiber types; 4) decrease in sensitivity to Ca(2+), particularly for SOL fibers (by approximately 40%); 5) decrease in the maximum steepness of the force-pCa curves, particularly for the hybrid I/IIA SOL fibers (by approximately 35%); and 6) increased occurrence of biphasic behavior with respect to Sr(2+) activation in SOL fibers, indicating the presence of both slow and fast troponin C isoforms. No fiber types common to the two muscles were detected at any time points (day 7, 21, and 50) after denervation. The results provide strong evidence that not only neural factors, but also the intrinsic properties of a muscle fiber, influence the structural and functional properties of a particular muscle cell and explain important functional changes induced by denervation at both whole muscle and single cell levels.

Animals↗