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Denervation increases a neurite-promoting activity in extracts of skeletal muscle.

During early stages of embryonic development, the motoneurones of the spinal cord send out axons that penetrate the differentiating muscle masses and establish connections with individual muscle fibres. It has been proposed that during this period the survival, differentiation and axon outgrowth of the motoneurones depend upon retrograde factors produced by the muscles, and in a previous study, we used a quantitative assay for neurite outgrowth from dissociated embryonic spinal neurones in vitro to characterize a neurite-promoting activity in media conditioned by embryonic muscle cells. At the adult neuromuscular junction, if some of the axons supplying a muscle are experimentally interrupted, fine nerve processes 'sprout' from the remaining intramuscular nerves and grow to innervate the denervated muscle fibres. In this situation also, it has been postulated that the denervated fibres release a diffusible sprouting stimulus. Using the same in vitro assay as before, we now report a striking increase of neurite-promoting activity in extracts of neonatal chick leg muscle after total denervation.

Animals↗

Autonomic denervation hypersensitivity in the primary glaucomas.

Autonomic denervation hypersensitivity of the iris was assessed in 44 patients with primary closed-angle glaucoma, 20 patients with primary open-angle glaucoma, and 40 age- and sex-matched control subjects. Significantly increased pupillary constriction in response to 2.5% methacholine chloride, indicating parasympathetic denervation hypersensitivity, was present in both closed-angle (p less than 0.02) and open-angle glaucoma patients (p less than 0.001), compared with the control group. Significantly increased pupillary dilatation in response to 0.5 per cent phenylephrine hydrochloride, indicating sympathetic denervation hypersensitivity of the iris, was also present in both closed-angle glaucoma (p less than 0.01) and open-angle glaucoma patients (p less than 0.05). The association between autonomic neuropathy and the primary glaucomas is discussed, with particular reference to the potential effects of autonomic dysfunction on aqueous dynamics.

Aged↗

Arpp, a new homolog of carp, is preferentially expressed in type 1 skeletal muscle fibers and is markedly induced by denervation.

In this study, we isolated and characterized a murine counterpart of the human Arpp (hArpp) gene. Sequence analysis revealed that the murine Arpp (mArpp) gene is almost identical to the Ankrd2 gene, which has recently been isolated as a mouse gene induced in stretched skeletal muscle. The mArpp gene encodes a protein of 332 amino acids that contains four well-conserved ankyrin-repeat domains in the central portion of the protein. The amino acid sequence of mArpp protein (mArpp) is highly homologous to that of mouse cardiac-restricted ankyrin-repeat protein (Carp), which is proposed to be a putative genetic marker for cardiac hypertrophy. Immunohistochemical analysis revealed that mArpp is preferentially expressed in type 1 skeletal muscle fibers, and that mArpp is localized in both the nucleus and the sarcomeric I-band of muscle fibers, suggesting that Arpp may function as a nuclear and sarcomeric protein. Furthermore, mArpp was also expressed in neurons of the cerebellum and cerebrum, the islets of Langerhans in the pancreas, and the esophageal epithelium, suggesting that mArpp may play a functional physiologic role in brain, pancreas, and esophagus as well as in type 1 muscle fibers. Interestingly, although mArpp was localized in both nucleus and cytoplasm in neurons, its localization was restricted to nucleus in pancreas and esophagus, suggesting that intracellular localization of mArpp is regulated in a tissue-specific manner. Furthermore, we found that mArpp- and Carp-expression in skeletal muscle were markedly up-regulated after denervation. Although the elevated expression level of Carp was kept only for two weeks after denervation, that of Arpp was kept at least for 4 weeks, suggesting that mArpp and Carp may play distinct functional roles in denervated skeletal muscle.

Animals↗

The effects of potassium ions and denervation on protein synthesis and the transport of amino acids in muscle.

1. The effects of varying concentrations of K(+) during incubation, of denervation and of various drugs on the accumulation of (14)C-labelled amino acids, their incorporation into protein and the stimulation of these processes by insulin in rat diaphragm preparations were studied. 2. The accumulation of glycine and aminoisobutyrate and incorporation of glycine into protein was less in tissue incubated in K(+)-free buffer or 20mm-K(+) than with 5-10mm-K(+). Incorporation of leucine was unaffected. 3. Incorporation into protein of amino acids by diaphragm that had been denervated 3 days previously was elevated. Accumulation of both glycine and aminoisobutyrate was also raised but that of phenylalanine was unaffected. 4. Accumulation of glycine by diaphragm and extensor digitorum longus muscle was decreased by a number of agents including cocaine and mepyramine. 5. The stimulation of incorporation by insulin was unaffected by changes in K(+) or in the presence of cocaine and mepyramine. Denervated tissue was markedly less responsive to insulin than its control. 6. The results are discussed in the context of the relation of amino acid accumulation to operation of the Na(+) pump and the influence of insulin thereon.

Acetylcholine↗

Effect of denervation on the synthesis of ribonucleic acid and deoxyribonucleic acid in rat diaphragm muscle.

1. The effect of unilateral denervation of rat diaphragm muscle on its content of nucleic acids and their incorporation of precursors was investigated. 2. After denervation the paralysed hemidiaphragm hypertrophies and within 3 days its content of RNA increases considerably. The concentration of DNA/unit mass remains fairly constant. 3. During this period there is some increase in the rate of incorporation of [(14)C]adenine into RNA, whereas there is some diminution in the rate of incorporation of [(14)C]orotic acid. 4. Incorporation of [(14)C]adenine and [(3)H]thymidine into DNA is much increased in the paralysed tissue, reaching its maximum by about the third day, but returning to normal by the tenth. 5. The significance of these results in relation to the hypertrophy after denervation is discussed.

Adenine↗

Effects of denervation on the glycogen content and on the activities of enzymes of glucose and glycogen metabolism in rat diaphragm muscle.

1. Changes in the content and concentration of glycogen and in the activity of a number of enzymes involved in glucose and glycogen metabolism were studied in the rat hemidiaphragm after unilateral denervation. 2. After nerve section the tissue hypertrophies; this hypertrophy is said to be confined to the smaller red fibres and not to the white. 3. The total hexokinase activity increases, whereas that of total glycogen phosphorylase decreases. The specific activity of phosphorylase a, determined after Halothane anaesthesia, remains fairly constant. 4. In fed animals the denervated tissue stores less glycogen, but in the early stages its glycogen content does not fall on starvation. 5. The effect of denervation on the specific activities of several other characteristically white-fibre enzymes are not consistent with the response of glycogen phosphorylase; the increase in content of glyceraldehyde 3-phosphate dehydrogenase and lactate dehydrogenase is thought to be related to proliferation of the sarcoplasmic reticulum. 6. The ratio of lactate dehydrogenase M/H subunits increases at the height of the hypertrophy, but then declines as the mass of the tissue falls. 7. The chronology of these changes in enzyme activities suggests a multiplicity of distinct responses after nerve section not consistent with any one model, either specific fibre development or reversion to de-differentiated, foetal-type metabolism.

Animals↗

The action of the beta-agonist clenbuterol on protein metabolism in innervated and denervated phasic muscles.

1. Clenbuterol treatment in innervated and denervated phasic extensor digitorum longus, plantaris and gastrocnemius muscles from rats caused a significant increase in RNA and protein contents in all muscles except denervated extensor digitorum longus. 2. All muscles showed an increase in the fractional rate of protein synthesis (Ks) with clenbuterol, but the temporal response varied. 3. The data suggest that the effect of clenbuterol on protein metabolism in innervated muscles is muscle-type specific, and demonstrate the homology of response for denervated muscles.

Animals↗

Reflex sympathetic dystrophy: result of autonomic denervation?

1. To investigate the nature of sympathetic dysfunction in the pathogenesis of reflex sympathetic dystrophy, the microcirculatory vasoconstrictive responses to dependency were investigated in the skin of the hand of 76 reflex sympathetic dystrophy patients with unilateral disease by means of laser Doppler flowmetry (in perfusion units) and capillary microscopy. The patients were divided into three stages according to their perception of skin temperature (stage I in the case of a stationary warmth sensation, stage II in the case of an intermittent warmth and cold sensation, and stage III in the case of a stationary cold sensation). The vasoconstrictive responses were induced by lowering of the affected hand. 2. As compared to controls, the mainly sympathetically mediated vasoconstrictive response at thermoregulatory level of the skin microcirculation, as measured by laser Doppler flowmetry, was attenuated at stage I (1.82 versus 1.41, P < 0.05), stage II (1.82 versus 1.09, P < 0.0001) and stage III (1.82 versus 1.14, P < 0.01), suggesting the involvement of sympathetic denervation at all stages of the reflex sympathetic dystrophy syndrome. This sympathetic denervation may also account for the observed increase in thermoregulatory skin blood flow at stage I as compared to controls (152 versus 81, P < 0.01). 3. Since sympathetic denervation has been reported to cause increased sensitivity of vascular structures to catecholamines, the decrease in thermoregulatory skin blood flow at stages II (54 versus 81, P < 0.05) and III (31 versus 81, P < 0.05), both as compared to controls, may result from hypersensitivity to catecholamines of skin microvessels. 4. The sympathetically independent vasoconstrictive response at the nutritive level of skin microcirculation, as measured by capillary microscopy, was impaired only at stage III as compared to controls (1.04 versus 2.06, P < 0.05). This divergence in microvascular reactivity upon dependency of the nutritive and thermoregulatory subsystems also supports the hypothesis of sympathetic dysfunction.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Disturbed trafficking of dystrophin and associated proteins in targetoid phenomena after chronic muscle denervation.

Dystrophin and associated proteins form a complex with an important role at the sarcolemma. Expression of this protein complex is highly regulated during development and regeneration. In order to better understand assembling patterns of these proteins, we have studied their expression in targetoid-like phenomena found in human muscle after chronic denervation, a situation known to give rise to abnormal protein trafficking. In eight biopsies of patients with chronic denervation, mainly resulting from amyotrophic lateral sclerosis, we found a number of targetoid phenomena. Selective accumulation of a number of sarcolemmal and sarcoplasmatic proteins occurred in targetoid phenomena. The larger majority of them contained gamma-sarcoglycan (gammaSG), but none contained the developmental heavy chain myosin isoform. In a series of 166 targetoid phenomena which could be studied with 17 different antibodies recognizing sarcolemmal and cytoplasmatic proteins, a high level of colocalization of gammaSG with desmin and alpha-actinin was found. Colocalization rate was much lower with other proteins, including other members of the dystrophin-associated protein complex. These data show that selective changes in expression of otherwise closely related proteins occur during disturbed trafficking leading to target formation. Because members of the dystrophin-associated protein complex do not accumulate in a similar fashion within targets, we suggest that a complex molecular control of gene expression and trafficking of this complex is involved after chronic muscle denervation.

Amyotrophic Lateral Sclerosis↗

Cellular changes in denervated tissue during wound healing in a rat model.

There is increasing evidence that innervation, possibly mediated via neuropeptides, promotes wound healing. This study presents data on the early cellular events during healing in denervated tissue. Free oblique groin flaps were raised on 25 adult Sprague-Dawley rats. Excisional wounds were placed within the flap and in two control sites, the contralateral inguinal region and the thorax. The absence of innervation in the free flap wounds was confirmed 10 days after surgery by indirect immunofluorescence with a pan-neuronal marker. The cellular infiltrate of the wounds was analysed immunohistochemically with a panel of antibodies to rat macrophages and monocytes (ED1), rat B lymphocytes (CD45R) and T lymphocytes (CD2). The immunostained cellular infiltrate was quantified at 2, 4, 7 and 10 days postoperatively. Our results show that denervated wounds have a significantly lower macrophage and T-lymphocyte count at day 4 of wound healing (P < 0.05). Inflammatory cells, particularly macrophages, are known to play an important part in wound healing and their reduced chemotaxis in denervated tissue may be related to the observed delay in wound closure.

Analysis of Variance↗

Rat adrenal transplants are reinnervated: an invalid model of denervated adrenal cortical tissue.

Adrenal autotransplantation is a widely used approach to investigate the potential for neural modulation of adrenal cortical function. It is believed that regenerating adrenal transplants are not reinnervated, thereby providing a model to investigate adrenal function in the absence of neural modulation. However, the hypothesis that adrenal transplants become reinnervated has not been directly tested. The purpose of the present study was to characterize the time course, extent, and nature of the reinnervation of the regenerating adrenal transplant and to assess whether the recovery of steroidogenic function and enzyme expression correlates temporally with the presence of innervation. Using immunohistofluorescent detection of tyrosine hydroxylase (TH), neuropeptide Y (NPY), calcitonin gene-related peptide (CGRP), and vasoactive intestinal peptide (VIP), the innervation of regenerating adrenals was assessed 14-30 days after transplantation of adrenal capsules beneath the kidney capsule in rats. Extensive reinnervation by TH-, NPY-, and VIP-positive fibres was present by 14 days after transplantation including regions of the adrenal capsule and cortex, with only minimal reinnervation by CGRP-positive fibres up to 30 days. TH- and NPY-positive chromaffin cells were also observed in the regenerating transplants. In addition, there was marked recovery of steroidogenic function and steroidogenic enzyme expression up to 30 days. The finding that nerve fibres are present in the transplants during the re-establishment of steroidogenic function and enzyme expression suggests that innervation may modulate the regeneration and functional recovery of adrenal transplants. In an attempt to prevent reinnervation of transplants, adrenal capsules were autotransplanted to denervated kidneys. Immunohistochemical analysis showed that, despite extensive denervation of the kidney tissue, the reinnervation and regeneration of the adrenal transplants still occurred. These data demonstrate the marked capacity of the regenerating adrenal to become reinnervated and reinforces the conclusion that adrenal transplants are an invalid model of denervated adrenal cortical tissue.

Adrenal Cortex↗

The incidence of post-vasectomy chronic testicular pain and the role of nerve stripping (denervation) of the spermatic cord in its management.

OBJECTIVE: To assess the incidence of chronic postvasectomy testicular pain (CPTP) and evaluate the use of denervation of the spermatic cord in its management. PATIENTS AND METHODS: A retrospective postal survey of 560 patients (mean age 36 years, range 25-55; mean time since vasectomy 19 months, range 8-39) who underwent vasectomy between July 1992 and December 1994 was carried out to determine the incidence of CPTP. A prospective study was conducted in a further group of 17 patients (mean age 43 years, range 34-60), who had had CPTP for at least one year, to evaluate the effectiveness of nerve stripping of the spermatic cord in relieving pain. RESULTS: Of 396 replies, 108 (27.2%) patients complained of some testicular pain following their vasectomy operation. In 88 (82%) of these 108 patients the pain was brief and was not defined as CPTP, while 20 (19%) patients had pain for > 3 months; 33 (31%) patients required analgesics to control the pain. Of the 17 patients who underwent spermatic cord denervation, 13 reported complete relief of pain at their first follow-up visit and were discharged. Four patients had a significant improvement in the symptom score and were satisfied with the results. CONCLUSIONS: There is a small but significant incidence of CPTP and patients should be warned of this possibility when counselled before operation. Denervation of the spermatic cord seems to be a viable surgical option for patients with CPTP who fail to respond to conservative measures.

Adult↗

Systemic administration of NMDA and AMPA receptor antagonists reverses the neurochemical changes induced by nigrostriatal denervation in basal ganglia.

In Parkinson's disease, nigrostriatal denervation leads to an overactivity of the subthalamic nucleus and its target areas, which is responsible of the clinical manifestations of the disease. Because the subthalamic nucleus uses glutamate as neurotransmitter and is innervated by glutamatergic fibers, pharmacological blockade of glutamate transmission might be expected to restore the cascade of neurochemical changes induced by a dopaminergic denervation within the basal ganglia. To test this hypothesis, two types of glutamate antagonists, the NMDA receptor antagonist MK-801 and the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptor antagonist LY293558, were administered systemically, either alone or in combination with L-DOPA, in rats with a unilateral 6-hydroxydopamine lesion of the nigrostriatal dopamine pathway. The effect of treatment was assessed neurochemically by analyzing at the cellular level the functional activity of basal ganglia output structures and the subthalamic nucleus using the expression levels of the mRNAs coding for glutamic acid decarboxylase and cytochrome oxidase, respectively, as molecular markers of neuronal activity. The present study shows that treatment with glutamate antagonists, and particularly with AMPA antagonists, alone or in combination with L-DOPA, reverses the overactivity of the subthalamic nucleus and its target areas induced by nigrostriatal denervation. These results furnish the neurochemical basis for the potential use of glutamate antagonists as therapeutic agents in Parkinson's disease.

Animals↗

Reserpine pretreatment prevents increases in extracellular striatal dopamine following L-DOPA administration in rats with nigrostriatal denervation.

The influence of L-DOPA and reserpine on extracellular dopamine (DA) levels in the striatum of intact and dopaminergic denervated rats was studied using the brain microdialysis technique. In intact rats, reserpine (5 mg/kg s.c.) reduced extracellular DA levels to 4% of basal values. L-DOPA (50 mg/kg i.p.) had no effect on extracellular DA levels in reserpine-pretreated rats. In rats with 6-hydroxydopamine-induced lesion of the nigrostriatal dopaminergic system, basal levels of extracellular DA were low but markedly increased by L-DOPA (50 mg/kg i.p.). In 6-hydroxydopamine-lesioned rats, pretreatment with reserpine (5 mg/kg s.c.) diminished L-DOPA (50 mg/kg i.p.)-induced increases in extracellular DA levels to 16% of those obtained in denervated animals not pretreated with reserpine (p<0.01). These results suggest that in the intact striatum, extracellular DA stems mainly from vesicular storage sites and that in the striatum with dopaminergic denervation, a large part of the L-DOPA-derived extracellular DA is also derived from a vesicular pool that is released by an exocytosis mechanism.

3,4-Dihydroxyphenylacetic Acid↗

Partial vagal denervation increases vulnerability to vagally induced atrial fibrillation.

INTRODUCTION: Cervical vagal stimulation shortens the atrial effective refractory period (ERP) primarily in the high right atrium (HRA) and facilitates induction of atrial fibrillation (AF) by single premature HRA extrastimuli. We hypothesized that vagal denervation of the HRA prevents both ERP shortening in the HRA and AF induction during vagal stimulation. METHODS AND RESULTS: Vagal denervation of the HRA was achieved using radiofrequency catheter ablation (RFA) of the fat pad at the right pulmonary vein-atrial junction (RPV fat pad). Programmed stimulation was performed at each of four atrial sites to measure ERP and inducibility of AF during vagal stimulation. RPV fat pad RFA increased only the HRA ERP during vagal stimulation (70 +/- 8.7 vs 117 +/-14.8, P < 0.05). RPV fat pad RFA increased measures of dispersion of refractoriness, the standard deviation of ERP (24 +/- 2.1 vs 33 +/- 2.0, P < 0.01), and the standard deviation of AF cycle length (11 +/- 0.8 vs 22 +/- 1.7, P < 0.001) during vagal stimulation. RPV fat pad RFA increased the incidence of AF (15/28 vs 24/28, P < 0.05) and the vulnerability (22 +/- 4.7 vs 39 +/- 5.6, P < 0.01) to AF induction during vagal stimulation, particularly from left atrial premature beats. After RPV fat pad RFA, premature beats induced AF by causing conduction block primarily in the HRA and macroreentrant activation around the block. CONCLUSION: Partial right atrial vagal denervation facilitated rather than prevented initiation of vagally mediated AF.

Adipose Tissue↗

[Subacromial denervation as a treatment modality for patients with subacromial pathology].

PURPOSE: The purpose of the presented study was to valuate the question whether subacromial denervation alone is sufficient in patients with subacromial pathology. MATERIAL AND METHODS: 20 patients (male: 7; female 13) with subacromial pathology without anterior acomionosteophytes, rotator cuff tear, calcific tendinitis or ac-joint pathology underwent endoscopically controlled subacromial denervation. The procedure was performed with an HF device. Patients were followed up at 6 weeks, 3 and 6 months with the Constant score. RESULTS: The preoperative Constant score was 59 +/- 18. Postoperatively the Constant score increased to 78 +/- 18 after 6 weeks, up to 83 +/- 5 after 3 months and up to 85 +/- 13 after 6 months. There was no difference between male and female patients nor between patients with different profession or athletic activity. We could not document any complication. Patients returned to work within 6 weeks after surgery. CONCLUSION: Patients without anterior acromion osteophytes and without rotator cuff tear may get good results with subacromial denervation alone and may not need acrornioplasty.

Arthroscopy↗

Effect of systemic creatine monohydrate supplementation on denervated muscle during reinnervation: experimental study in the rat.

The purpose of this experimental study was to evaluate possible upgrading effects of systemic creatine monohydrate administration on the reinnervation of denervated muscle. At the same time, the protective effect of the agent on denervated muscle until ultimate reinnervation after nerve repair was quantified. The functional outcome of muscle reinnervation after creatine monohydrate application was compared with a control group. Forty adult Wistar rats weighing 180 to 220 g were used. The right sciatic nerve was dissected, exposed, and cut at the level of the midthigh in all rats. The experimental design consisted of two groups: experimental (animals were fed creatine monohydrate) and control (gavage feeding was provided by saline). Both groups were divided into two subgroups: subgroups A and B for the experimental group, and subgroups C and D for the control group. In subgroups A and C, the nerves were repaired with four 10-0 epineurial stitches. In subgroups B and D, both the proximal and distal ends of the nerves were ligated and no neural anastomosis was performed. In the experimental groups (subgroups A and B), the rats were fed by daily supplementation of oral creatine monohydrate, 300 mg/kg body weight. In the controls (subgroups C and D), oral supplementation was provided by saline. Functional recovery was evaluated using walking track analysis, pinching test, and limb circumference and toe contracture measurements at the end of 6 months, after which the rats were sacrificed and nerve specimens from both ends of the repair sites and the whole gastrocnemius muscle were obtained to document the results of the histomorphometric and histochemical studies, including light microscopic examinations and muscle weight measurements. The mean functional recovery values in subgroups A, B, C, and D were 91 percent, 80 percent, 87 percent, and 59 percent, respectively. Functional recovery improved significantly in the experimental groups (in both the surgically repaired and unrepaired subgroups), compared with the control groups (p<0.05). The pinching test revealed a statistically significant difference in nerve conduction between the experimental and control groups (p<0.05). The limb circumference ratio of the surgically treated side to the untouched side in subgroups A, B, C, and D were noted as 0.95, 0.89, 0.91, and 0.87, respectively, and the difference between the experimental and the control groups was statistically significant (p<0.05). The differences between subgroups A and B, C and D, A and C, and B and D were also significant. The surgically repaired and creatine-supplemented subgroups demonstrated the best results in toe contracture index. The muscle weight measurement results were concordant with the results of the limb circumference ratio. In both surgically repaired subgroups (subgroups A and C), there were qualitatively significant amounts of myelinated fibers in the nerve distal to the anastomotic site; there were no myelinated fibers in the distal stumps of subgroups B and D. Histochemical analyses of the contents of the muscle fiber types also revealed no significant difference. Overall, the results showed the useful effect of oral creatine supplementation on both surgically repaired and unrepaired nerve injuries. The best results were obtained from surgically repaired nerve injuries and also from the systemic creatine-supplemented subgroups. This study confirms that systemic administration of creatine monohydrate has a protective and upgrading effect on the functional properties of denervated muscle, especially in surgically reinnervated subjects.

Animals↗

Preventing denervation atrophy of a grafted muscle.

Electrodes were implanted in the grafted muscles of rabbits to generate continuous stimulation for the purpose of preventing denervation atrophy. Denervation atrophy could be prevented, to some extent, in dissected muscles with vascular pedicles not occluded during the experiment. Denervation atrophy was found to be promoted, rather than prevented, in grafted muscles with occluded vascular pedicles. The crucial conditions are varying stimulation and the long duration of occlusion (90 min). Overly strong stimulation of muscle contraction appears to result in "fatigue" of grafted muscles in which blood circulation has been blocked for more than the 90 min used in the reported experiment. Further investigations are necessary of the conditions of stimulation and the point in time at which stimulation should commence after grafting.

Animals↗