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PubMed1989

Interaction between clonidine and physostigmine in normal rats and in rats after sinoaortic denervation.

Clonidine (3-30 micrograms.kg-1, i.v.) induced a fall in mean arterial pressure in rats after sinoaortic denervation but not in sham-operated animals. Moreover, sinoaortic denervation reduced the bradycardic action of this antihypertensive drug. Pressor and tachycardic response to physostigmine (60 micrograms.kg-1, i.v.) were greater in denervated than in sham-operated rats. The increase of mean arterial pressure was 26.2 +/- 2.2 mm Hg in sham-operated rats (n = 12) and 53.8 +/- 2.0 mm Hg in denervated rats (n = 12, P less than 0.005). Pretreatment with 3 micrograms.kg-1 (i.v.) of clonidine did not alter the pressor response to physostigmine (60 micrograms.kg-1) in either of the two groups; 10 and 30 micrograms.kg-1 of clonidine reduced the physostigmine-induced increase of mean arterial pressure in sham-operated rats but enhanced the pressor response in denervated animals. Furthermore, an ineffective dose of physostigmine (30 micrograms.kg-1, i.v.) induced a pressor response after pretreatment with clonidine (10 micrograms.kg-1) in denervated rats. Clonidine (10 micrograms.kg-1) did not affect the pressor effect of 1,1 dimethyl-4-phenylpiperazinium iodide (DMPP: 50 micrograms.kg-1, i.v.) or phenylephrine (4 micrograms.kg-1, i.v.) in either group. The anticholinergic effect of clonidine in sham-operated rats may be explained by an inhibitory action on the release of acetylcholine in several brain structures but the facilitatory effect of clonidine observed in denervated animals is not clear. The results did not suggest a peripheral involvement in this facilitatory effect.

Animals↗
PubMed1984

Lysosomes in skeletal muscle following denervation. Time course of horseradish peroxidase uptake and increase of lysosomal enzymes.

The in-vivo uptake of exogenously applied horseradish peroxidase and the activities of the lysosomal enzymes acid phosphatase and cathepsin D were studied histochemically and/or biochemically in innervated and 2-14 day-denervated tibialis anterior muscles of the mouse. The biochemically determined uptake of horseradish peroxidase showed a large increase already 4 days after denervation. The activities of the lysosomal enzymes increased in a more gradual fashion, and only cathepsin D showed an increase in activity when expressed as total activity per muscle. Histochemically horseradish peroxidase was found to be localized in muscle fibres in characteristic spindle-shaped segments after denervation. The main increase in the number of such segments per transverse section of the muscle occurred between 3 and 6 days after denervation. In serial sections these segments frequently showed positive staining also for acid phosphatase. It is concluded that exogenously applied horseradish peroxidase is taken up into the lysosomal system, which after denervation becomes organized into characteristic spindle-shaped segments in the muscle fibres. The endocytic activity of muscle fibres increases early after denervation. This is followed by a more gradual increase in activity of lysosomal enzymes and finally by an organization of the lysosomal system into characteristic spindle-shaped segments. The results are compatible with the working hypothesis that increased endocytosis may initiate lysosomal activation in denervated skeletal muscle.

Acid Phosphatase↗
PubMed1987

Hypertrophy of radial and ulnar arteries following denervation of the chicken wing.

Denervation of radial and ulnar arteries in the growing and adult domestic fowl was achieved by unilateral sectioning of the brachial plexus. Eight weeks later the denervated arteries and those of the contralateral wing were examined with light- and electron microscopy to determine the effect of denervation on arterial structure. In growing fowls, the area of the media in radial and ulnar arteries was increased by 29% and 25%, respectively, after denervation. The number of smooth muscle layers was also significantly increased by 16% (radial) and 14% (ulnar), but no significant variation was seen in the wall/lumen ratio of either growing artery. In adult fowls, the area of the media was increased by 93% (radial) and 32% (ulnar) following denervation and the number of smooth muscle cell layers increased by 39% (radial) and 11% (ulnar). There was also an increased wall/lumen ratio of 64% (radial) and 92% (ulnar). These results indicate that hyperplasia of smooth muscle has occurred in response to denervation. Flow-cytometric DNA analysis of growing arteries also indicates that the increase in muscle-cell volume is a result of cell division (not polyploidy) since no significant differences were found between the control and denervated arteries in any stages of the cell cycle.

Animals↗
PubMed1982

Inhibition of insulin-stimulated xylose uptake in denervated rat soleus muscle: a post-receptor effect.

Insulin (100 U/l) stimulated xylose uptake in rat soleus muscle from a basal value of 2.3 +/- 0.5 to 11.6 +/- 2.1 mumol . g-1 . h-1. Denervation (section of the sciatic nerve) markedly reduced the stimulatory action of insulin (basal 1.3 +/- 0.4 mumol . g-1 . h-1; insulin-stimulated 4.5 +/- 0.6 mumol . g-1 . h-1). This effect appeared 3 days after denervation and was maximal after 5 days. Denervation also affected the insulin dose response curve; there was no effect of insulin in denervated muscle until the concentration exceeded 1 U/l. Denervation inhibited insulin-stimulated alpha-aminoisobutyrate uptake by 77% but did not affect 125I-insulin binding or glucose-independent activation of glycogen synthase by insulin. There was no effect of denervation on the insulinomimetic effects of concanavalin A, hydrogen peroxide, vitamin K5, anoxia, 2:4-dinitrophenol, cooling, hyperosmolarity or EDTA, but the effect of diamide was inhibited. It is concluded [1] that denervation inhibits insulin-stimulated sugar transport at some early post-receptor step, and [2] that the mechanism whereby insulin activates glycogen synthase is different from the activation of the membrane transport of sugars and amino acids.

Aminoisobutyric Acids↗
PubMed1984

Gangliosides in rabbit and human skeletal muscle with denervation atrophy.

In experimental denervation of rabbit skeletal muscle, an increase of gangliosides has been reported. Ganglioside, DNA, total lipid and protein levels were studied in experimental denervation of rabbit gastrocnemius muscle and in human skeletal muscle with denervation atrophy. In severe human denervation atrophy total ganglioside, DNA and total lipid levels were increased, and protein levels decreased as compared with controls. The ganglioside pattern was changed in severe human denervation atrophy and showed a significant increase of a GT1b-like and moderate decrease of a GM1-like compound. Findings in experimentally denervated rabbit muscle resembled results obtained in human denervation atrophy, showing a significant increase of total ganglioside levels as well as a moderate increase of GT1b-like and decrease of GM1-like compounds.

Adult↗
PubMed1993

Protein secretion from mouse skeletal muscle: coupling of increased exocytotic and endocytotic activities in denervated muscle.

The secretion of proteins labelled by incorporation of radioactive amino acids was studied in innervated and 10 to 13-day-denervated mouse skeletal muscle. The secretion of 3H-leucine-labelled proteins, expressed per mg muscle wet weight, increased after denervation, and the kinetics of the secretory process was also altered in denervated muscle. Separation of secreted 35S-methionine-labelled proteins by sodium dodecyl sulphate polyacrylamide gel electrophoresis followed by autoradiography revealed some denervation-induced alterations in the pattern of secreted proteins. The secretion from both innervated and denervated muscle was highly temperature sensitive and was reversibly inhibited by brefeldin A, a drug that blocks forward membrane transport from the endoplasmic reticulum/Golgi apparatus. This drug was also found to inhibit the uptake of fluorescein isothiocyanate-labelled dextran in denervated muscle but had no effect on the endocytotic activity of innervated muscle. This lends support to the hypothesis that the increased endocytotic activity in denervated muscle is coupled to a high secretory activity.

Animals↗
PubMed1994

Effect of unilateral surgical denervation of brown adipose tissue on uncoupling protein mRNA level and cytochrom-c-oxidase activity in the Djungarian hamster.

The bilateral lobe of interscapular brown adipose tissue of the Djungarian hamster was unilaterally denervated in order to study the role of the sympathetic innervation for maintenance and cold-induced increase of non-shivering thermogenesis. Denervation decreased the noradrenaline content of brown adipose tissue to less than 9% of the intact contralateral pad. This low noradrenaline level was maintained for 1-14 days after denervation. First, to study the role of the sympathetic innervation of brown adipose tissue in the maintenance of the high thermogenic capacity characteristic of the cold acclimated state, brown adipose tissue was denervated in hamsters either kept at thermoneutrality or acclimated to 5 degrees C ambient temperature for 4 weeks. Cold-acclimated hamsters had elevated levels of uncoupling protein messenger ribonucleic acid (8.1-fold) and cytochrom-c oxidase-activity (3-fold). Denervation of brown adipose tissue decreased uncoupling protein-messenger ribonucleic acid level and cytochrom-c-oxidase-activity as compared to the intact pad in thermoneutral and in cold-acclimated hamsters. However, in cold-acclimated hamsters uncoupling protein-messenger ribonucleic acid level and cytochrom-c-oxidase-activity in denervated brown adipose tissue both were maintained on an elevated 6-fold higher level as compared to thermoneutral controls. Second, to study the role of the sympathetic innervation of brown adipose tissue in the cold-induced increase in thermogenic capacity, hamsters were denervated prior to cold acclimation and responses were measured after 3 and 14 days of cold exposure. Uncoupling protein-messenger ribonucleic acid level and cytochrom-c-oxidase-activity of intact brown adipose tissue increased after 14 days cold acclimation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adipose Tissue, Brown↗
PubMed1994

Jun, Fos, MyoD1, and myogenin proteins are increased in skeletal muscle fiber nuclei after denervation.

After denervation, mRNA levels of the jun and fos protooncogenes and of the muscular differentiation factors myoD1 and myogenin are increased. Here, immunohistochemistry was used (a) to show that this increase in mRNA is followed by an increase in the transcription factor proteins, and (b) to determine which cell populations in skeletal muscle express these factors after denervation. Rat diaphragms were denervated and analyzed after periods of 90 min-8 days. An increase in Fos and Jun as well as MyoD1 and Myogenin immunoreactivity was found after 2-2.5 days of denervation. Fos, MyoD1, and Myogenin immunoreactivity was mostly confined to muscle cell nuclei, whereas Jun antibodies stained muscle cell and some interstitial cell nuclei. A selective expression of any of the four transcription factors in muscle cell nuclei closely associated with motor endplates could not be detected in either denervated or innervated muscle at any time point examined, indicating that synaptic and extrasynaptic muscle cell nuclei are activated simultaneously after denervation. These results suggest that a genetic program which includes protooncogenes and myogenic differentiation factors is activated in skeletal muscle after denervation.

Amino Acid Sequence↗
PubMed1985

Subsensitivity of enteric cholinergic neurones to alpha 2-adrenoceptor agonists after chronic sympathetic denervation.

The concentration-effect relationships of noradrenaline, dopamine and clonidine in inhibiting resting and stimulated acetylcholine output have been studied in intact and in sympathetically denervated preparations of guinea pig isolated distal colon. The order of potencies for the inhibition of resting acetylcholine release in intact preparations was clonidine greater than dopamine greater than noradrenaline while the order of intrinsic activities was noradrenaline greater than dopamine greater than clonidine. Sympathetic denervation was able to modify the potency of either clonidine, dopamine and noradrenaline. Noradrenaline was 6 times more potent in inhibiting resting acetylcholine release in denervated than in intact preparations, while clonidine and dopamine underwent a 18-fold and a 11-fold decrease in potency after denervation. The potency of clonidine relative to noradrenaline was 110 in intact preparations and only 1.2 in denervated organs. The intrinsic activities of noradrenaline, dopamine and clonidine were almost unchanged in denervated organs. A dose-dependent facilitatory effect of yohimbine on both the resting acetylcholine output and the peristaltic reflex could be observed in intact but not in sympathetically denervated preparations at concentrations ranging from 2.5 X 10(-8) M to 2.5 X 10(-7) M. Yohimbine was able to counteract the inhibitory effect of dopamine and to remove the inhibitory effect of periarterial nerve stimulation on both acetylcholine release and the peristaltic reflex. Our results are consistent with the existence of a tonic physiological modulation of enteric cholinergic neurones by postganglionic sympathetic fibres. The order of potencies of adrenoceptor agonists and the antagonism by yohimbine is consistent with such a modulation being entirely carried out through alpha 2-heteroceptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗
PubMed1981

Changes in mechanical properties of the inferior oblique muscle of the rabbit after denervation.

The inferior oblique muscle (IO) of the rabbit was denervated. The mechanical properties of the muscles were determined at 35 degrees C in vitro 2--76 days after the operation. After denervation the muscles exhibited a considerable hypertrophy. The cross-sectional area of muscles denervated 30 days or longer grew to about twice as innervated controls. The length-tension relationship of passive or activated long-term denervated IO differed from normal in a higher stiffness. After denervation the time parameters of single twitches and tetanic contractions evoked by massive stimulation were prolonged, the fusion frequency was decreased, and the fatigue resistance was increased. During the first week after cutting the motor nerve both twitch and tetanic tensions decreased drastically. A minimum was reached at the end of the second weak. In the subsequent weeks the tension development was increased again, but per unit cross-sectional area it was always smaller than in innervated muscles. After denervation the twitch:tetanus ratio was increased. Cooling to 25 degrees C was followed by an increase in time parameters of single twitches and tetanic contractions and by a depression of twitch and tetanic tensions. Following a repetitive stimulation denervated IO showed a posttetanic depression of the single twitch.

Animals↗
PubMed1984

Lysosomal and energy enzyme activities in hypertrophied rat soleus muscle after denervation.

Muscle hypertrophy was induced in the soleus muscle of young rats by tenotomy of the gastrocnemius and plantaris muscles. Three and 7 days afterwards the sciatic nerve was sectioned. The loss of weight of muscles subjected to this combined procedure three days after denervation was 30-40%. Lysosomal enzyme activities (acid phosphatase, alpha-glucosidase, beta-galactosidase and N-acetyl-beta-D-glucosaminidase) and energy enzyme activities (lactate dehydrogenase, LDH, triose-3-phosphate dehydrogenase, TPDH , D-hexokinase, HK and citrate synthase, CS) were determined 3 days after denervation, 3, 7 and 10 days after hypertrophy had been induced and 3 days after denervation of hypertrophying muscles on day 3 and 7. Normal non-operated rats of corresponding body weight served as controls and their enzyme activities were estimated on the same day. In the course of muscle hypertrophy, the 4 lysosomal enzyme activities increased progressively. Although 3 days' denervation of control muscles did not alter lysosomal enzyme activities, denervation of hypertrophying muscles greatly enhanced the activity of these enzymes. Enzymes of energy metabolism were affected to a lesser degree. The results suggest that denervation of hypertrophying muscles causes more extreme changes in muscle weight and lysosomal enzyme activities than denervation alone. The possible implications of this finding are discussed in relation to the rapid atrophy.

Animals↗
PubMed1987

Effects of low and high frequency patterns of stimulation on contractile properties, enzyme activities and myosin light chain accumulation in slow and fast denervated muscles of the chicken.

The effects of denervation and direct stimulation in fast and slow latissimus dorsii muscles were investigated in chicken. In slow ALD muscle, denervation resulted in an incompleteness of the relaxation, a decrease in MDH and CPK activities and an increase in fast myosin light chains (MLC) accumulation. Direct stimulation at either fast or slow rhythm prevented the effects of denervation on relaxation and CPK activity but was ineffective on MDH activity and fast MLC accumulation. Moreover, direct stimulation of denervated ALD caused rhythm-dependent change in tetanic contraction. In fast PLD muscle, the main changes in muscle properties following denervation were a slowing down of the time course of the twitch and an incompleteness of the relaxation, a decrease in LDH and CPK activities and in LC3F accumulation. Stimulation at a high frequency partly prevented the effects of denervation and resulted in a large accumulation of LC3F, while a low frequency stimulation did not restore the twitch time to peak, increased MDH activity and induced synthesis of slow MLC. This study emphasizes the role of muscle activity and its pattern in some properties of slow and fast chicken muscles following denervation.

Animals↗
PubMed1981

Changes in lipid levels of three skeletal muscles following denervation.

Nonpolar and polar lipids extracted from denervated rat gastrocnemius, plantaris, and soleus muscles were measured 7-9 days after unilateral sciatic nerve transection. The contralateral muscle (CCON) was used to obtain control lipid levels. After denervation changes in lipid concentrations were found in all three muscles. These alterations in lipid levels were generally in same direction but not to the same extent. The change in total nonpolar lipids (NL) was an increase in soleus greater than gastrocnemius greater than plantaris concentration. This change in lipid concentration was more apparent than real since the wet weight of muscle was decreased after denervation. Since polar lipid (PL) concentrations were not increased under these conditions of muscle weight loss, an actual decrease of polar lipids after denervation may be inferred. In contrast to the other two muscles, a marked difference was noted for polar lipids of denervated gastrocnemius muscle. An unidentified spot near the origin was detected. This area is the location of a nerve sprouting factor(s). The compound(s) was not detectable for the other two muscles. When the gastrocnemius from an unoperated animal rather than a CCON muscle was used as a benchmark, slight increases were found for total nonpolar, polar, and plasmalogen fractions following denervation. The changes for individual lipid fractions were less definable, except for the significant increase for the unknown polar compound near the origin. This spot was noted in extracts from CCON and DEN muscles but not in untouched control muscle. The CCON gastrocnemius muscle is therefore a poor control for determining effects of denervation on lipid levels and perhaps other biochemical parameters as well.

Animals↗
PubMed1991

Effects of denervation on the distribution of peanut agglutinin binding molecules in frog muscles.

The extracellular matrix has been shown to play an important role in the differentiation of neuromuscular junctions during reinnervation in frogs. Peanut agglutinin, a lectin, is known to specifically bind to some glycoconjugates in the extracellular matrix at the frog neuromuscular junction and myotendinous junction. In order to determine if innervation has any role in regulating the specific binding of peanut agglutinin at neuromuscular junctions and myotendinous junctions, the distribution of peanut agglutinin binding was examined in muscles chronically denervated for various periods. Short-term denervated muscles (less than or equal to 2 months) showed no changes in peanut binding agglutinin binding at neuromuscular junctions and no extrajunctional binding. In contrast, long-term denervation (greater than 2 months - 7.5 months) resulted in altered peanut agglutinin distribution and a substantial reduction or a total loss in its binding at denervated neuromuscular junctions; binding at myotendinous junctions was not affected. Results of electron microscopic studies suggest that the presence of Schwann cells at denervated endplates delays the loss of peanut agglutinin binding. Reinnervation restores normal peanut agglutinin binding at neuromuscular junctions following long-term denervation. This report demonstrates that although the distribution of peanut agglutinin binding molecules is unchanged short-term denervation, intact innervation is necessary for the long-term maintenance of these molecules at neuromuscular junctions.

Animals↗
PubMed1991

A comparison of fibrillation in denervated skeletal muscle of the anaesthetized rat and guinea-pig.

We have used intracellular recording in vivo to study fibrillation (spontaneous repetitive membrane activity) in extensor digitorum longus (fast twitch) and soleus (slow twitch) muscles of the anaesthetized rat and guinea-pig denervated for periods of about 10 to 60 days. The proportion of fibres fibrillating in the guinea-pig soleus was greater than 50% in most animals up to the longest period of denervation (65 days). Fibrillation was rarely found in rat soleus after three weeks of denervation. Its incidence in the extensor digitorum longus muscles of both species was intermediate. The mean frequency of fibrillation was higher in guinea-pig extensor digitorum longus (16 Hz) and soleus (8 Hz) than in the rat extensor digitorum longus (3 Hz) and soleus (2 Hz). The resting membrane potentials of the denervated muscles were less than normal and correlated inversely with the frequency of fibrillation but not with the incidence of fibrillation: in rat soleus, many fewer fibers were fibrillating at a given membrane potential than in the other three muscles. The incidence of fibrillation was compared with previously reported tensions of the four denervated muscles and was found to have the same rank order. We suggest that fibrillation may reduce atrophy (and hence tension loss) of denervated muscle, which may have implications for artificial stimulation. Fibrillation frequency was directly related to changes in twitch speed of the four muscles after denervation.

Animals↗
PubMed1988

Differentiation of fast and slow muscles in the rat after neonatal denervation: a physiological study.

Whether an intact innervation is essential for postnatal muscle differentiation was examined in the rat by recording physiological contraction parameters. Muscles in one leg were denervated neonatally (within 24 h of birth) and, between 3-28 days after the operation, their contractions were compared with those of the contralateral control muscles. Experiments were performed on the extensor digitorum longus (edl, a fast muscle) and the soleus (a slow muscle) muscles and contractions were recorded in vitro, at 35 degrees C and with direct stimulation. When compared with the control muscles, 3-4-day-old neonatally denervated fast and slow muscles had longer twitch contractions, higher twitch/tetanus ratios and certain other specific differences in their contraction parameters. These denervation-induced changes in neonatal muscles were essentially similar to those produced 3-7 days after denervation in the differentiated (4-week-old) fast muscle. Despite differences in their absolute values, the contraction parameters of neonatally denervated and control edl muscles changed similarly during development, indicating that postnatal differentiation of fast muscle fibres is independent of a neuronal influence. In the case of the neonatally denervated soleus muscle, the developmental changes in contraction parameters, i.e. shortening of the twitch duration, increase of rate of rise and rate of relaxation in the tetanus and increase of the maximum shortening velocity, were more pronounced than in the control slow muscle; also, there were similarities with the pattern of fast muscle differentiation. Thus, muscle fibre differentiation in soleus becomes altered towards that of a fast muscle after neonatal denervation.

Age Factors↗
PubMed2002

Protective effects of ciliary neurotrophic factor on denervated skeletal muscle.

To study the effects of ciliary neurotrophic factor (CNTF) on denervated skeletal muscle atrophy and to find a new approach to ameliorate atrophy of denervated muscle, a model was established by cutting the right sciatic nerve in 36 Wistar mice, with the left side serving as control. Then they were divided into two groups randomly. CNTF (1 U/ml) 0.1 ml was injected into the right tibial muscle every day in experimental group, and saline was used into another group for comparison. The muscle wet weight, muscle total protein, Ca2+, physiological response and morphology were analyzed on the 7th, 14th and 28th day after operation. Our results showed that compared to control group, there was a significant increase in muscle wet weight, total protein, Ca2+, muscle fiber cross-section area in CNTF group (P < 0.05). CNTF could ameliorate the decrease of tetanic tension (PO), post-tetanic twitch potentiation (PTP), and the prolonged muscle relaxation time (RT) caused by denervation (P < 0.05). The motor end-plate areas 7 days and 14 days after denervation was similar (P > 0.05), but significantly larger 28 days after the denervation (P < 0.05). Our results suggest that CNTF exerts myotrophic effects by attenuating the morphological and functional changes associated with denervation of rat muscles and has protective effects on denervated muscle and motor end plate.

Animals↗
PubMed2003

Functional characterisation of alpha(1)-adrenoceptors in denervated rat vas deferens.

We investigated whether or not surgical denervation of the rat vas deferens changes the alpha(1)-adrenoceptor subtypes involved in the contractions to noradrenaline. Denervated vas deferens was approximately 22 times more sensitive to noradrenaline (pD(2)=7.35+/-0.04) than control vas (pD(2)=6.01+/-0.03). This difference in noradrenaline potency was eliminated when cocaine (6 microM) was added to control vas (pD(2)=7.22+/-0.04). The noradrenaline-induced contractions of control and denervated vas deferens were insensitive to the alpha(1B)/alpha(1D)-adrenoceptor alkylating agent chloroethylclonidine (100 microM, 45 min). The concentration-response curves to noradrenaline in control and denervated vas deferens were competitively antagonised by prazosin (pA(2) approximately equal to 9.6), WB-4101 (pA(2) approximately equal to 9.5), 5-methyl urapidil (pA(2) approximately equal to 8.4), phentolamine (pA(2) approximately equal to 8.7), yohimbine (pA(2) approximately equal to 6.9), BMY 7378 (pA(2) approximately equal to 6.9) and indoramin (pA(2) approximately equal to 8.7). After the treatment of control and denervated vas deferens with phenoxybenzamine, the partial agonist oxymetazoline antagonised competitively the concentration-response curves to noradrenaline showing pA(2) values approximately 7.4 in both groups. We conclude that noradrenaline-induced contractions in control and denervated rat vas deferens are mediated by alpha(1A)-adrenoceptors and that surgical denervation of the rat vas deferens is not able to change the alpha(1)-adrenoceptor subtypes involved in the contractions to noradrenaline.

Adrenergic alpha-Agonists↗