Effects of short-term denervation and subsequent reinnervation on motor endplates and the soleus muscle in the rat

The rat sciatic nerve was locally frozen, and changes in the nerve, motor endplates, and the soleus muscle were examined for up to 6 weeks by light and electron microscopy. The wet weights of denervated soleus muscles compared with contralateral values progressively declined to a minimum at 2 weeks after injury (60.7 +/- 2.5%) and began to reverse following 3 weeks. The sciatic nerve thoroughly degenerated after freezing. However, numerous regenerated myelinated and thin nerve fibers were observed at 3 weeks. They were considerably enlarged but still smaller than normal counterparts at 6 weeks postoperatively. Nerve terminals containing synaptic vesicles of endplates disappeared at day 1 and mostly reappeared at 3 weeks (about 70% of the endplates). All endplates examined were reinnervated at 4, 5, and 6 weeks. On the other hand, postsynaptic folds of muscle fibers seemed to be only slightly influenced by denervation or reinnervation. Ultrastructural alterations of myofibrils, in particular the loss of register, immediately appeared after denervation, spread progressively, peaked at 2 weeks, ameliorated following reinnervation, and became significantly normalized at 6 weeks after freezing. The proportion of type II fibers in the soleus muscle similary showed an increase and a decrease with a short delay in response to denervation and reinnervation, respectively. This study clearly demonstrated that the nerve supply affects the ultrastructural integrity of skeletal muscles. In addition, changes in the endplates and the soleus muscle evaluated in this study after short-term denervation are largely reversible following reinnervation.     

The influence of secretin on the secretion of pepsin in response to acid stimulants in the anaesthetized cat.

1. The problem of selectivity during reinnervation of skeletal muscle fibres was investigated in the rat using the fast-twitch extensor digitorum longus (EDL) and the slow-twitch soleus muscles and their nerves. 2. After an operation on these nerves permitting them to compete for reinnervation of one or the other muscle (hereafter called Y-union), virtually the total isometric tetanic tension of EDL muscle could be elicited by stimulating the EDL nerve, while stimulating the soleus nerve yielded little or no tension. In the case of the soleus muscle, stimulation of either nerve elicited about half of the total isometric tetanic tension. 3. During the course of reinnervation of these muscles in non-competitive situations, the time course of increase in the ratio of tension elicited by nerve stimulation to that by direct stimulation was slower in the case of soleus nerve reinnervating EDL muscle, compared with cross-reinnervation in the reverse direction or reinnervation of each muscle by its own nerve. 4. Crushing the common peroneal nerve 12 days after a Y-union in an attempt to retard the EDL nerve did not favour reinnervation of the EDL by soleus nerve, but crushing the nerve again or just once at 1 month after the original operation produced substantial partial reinnervation of the EDL by the soleus nerve. 5. It is concluded that soleus nerve fibres form functioning neuromuscular synapses on EDL muscle fibres only with difficulty. The pattern of reinnervation reveals characteristic differences between fast-twitch and slow-twitch muscles on the one hand and between their respective nerves on the other.     

Long-term retention of regenerative capability after denervation of skeletal muscle, and dependency of late differentiation on innervation

The present study examines the influence of denervation on the regenerative ability of skeletal muscle in rats. Muscle denervation was achieved by transecting and ligating the cut ends of the sciatic nerve. Four to 48 weeks after denervation, the extensor digitorum longus (EDL) muscle was autotransplanted to induce muscle regeneration. The transplanted EDL muscles were examined at 1-12 weeks. Normal (i.e., no prior denervation) EDL muscle autotransplants were also examined for comparison. Denervation resulted in progressive atrophy of muscle, marked by a reduction in the size of myofibers and an increase in endomysialperimysial connective tissue. In spite of these alterations, typical events of muscle regeneration were invariably observed after transplantation. Initial myofiber degeneration and subsequent regeneration of myotubes occurred in a manner similar to normal muscle transplants. However, only a partial maturation of myotubes was observed in denervated muscles. These results show that extended denervation does not abolish the capability for muscle regeneration. The precursor myosatellite cells, proposed to be responsible for muscle regeneration, retain their regenerative potential after denervation. It is concluded, however, that the presence of intact innervation is crucial for the terminal differentiation and maturation of regenerating muscle.     

Long-term reiention of regenerative capability after denervation of skeletal muscle,and dependency of late differentiation on innervation

The present study examines the influence of denervation on the regenerative ability of skeletal muscle in rats. Muscle denervation was achieved by transecting and ligating the cut ends of the sciatic nerve. Four to 48 weeks after denervation, the extensor digitorum longus (EDL) muscle was autotransplanted to induce muscle regeneration. The transplanted EDL muscles were examined at 1–12 weeks. Normal (i.e., no prior denervation) EDL muscle autotransplants were also examined for comparison. Denervation resulted in progressive atrophy of muscle, marked by a reduction in the size of myofibers and an increase in endomysialperimysial connective tissue. In spite of these alterations, typical events of muscle regeneration were invariably observed after transplantation. Initial myofiber degeneration and subsequent regeneration of myotubes occurred in a manner similar to normal muscle transplants. However, only a partial maturation of myotubes was observed in denervated muscles. These results show that extended denervation does not abolish the capability for muscle regeneration. The precursor myosatellite cells, proposed to be responsible for muscle regeneration, retain their regenerative potential after denervation. It is concluded, however, that the presence of intact innervation is crucial for the terminal differentation and maturation of regenerating muscle.     

Reinnervation and recovery of mouse soleus muscle after long-term denervation

Reinnervation and recovery of the mouse soleus muscle were studied 2-10 months after denervation periods of about 7 months. To maintain denervation the right sciatic nerve was frozen 14 times at 2-week intervals. Though initially intermittent muscle reinnervation occurred, contractile force of denervated muscles was reduced to less than 10% of the contralateral muscles by the fifth nerve freezing and further declined thereafter. Following reinnervation, recovery of soleus muscle force proceeded slowly to reach plateau values after 5-6 months. Tetanic muscle force reached on average 72% (range 58-86%, n = 12) of contralateral muscles after 5-10 months, (P less than 0.01, t-test for absolute values) and 87% of unoperated animals after 10 months (P less than 0.05, n = 5). Muscle fibre diameters were significantly reduced in reinnervated muscles, but frequency distributions were normal and similarly shaped in reinnervated and control muscles, suggesting complete muscle reinnervation and the absence of denervated fibres even at 2 months of reinnervation. Total numbers of muscle fibres were similar in reinnervated (842 +/- 73 S.D., n = 15), contralateral (854 +/- 104 S.D., n = 15) and control soleus muscles (853 +/- 77 S.D., n = 5). The number of myelinated axons in regenerating soleus nerves reached control values by 3 months after the last freezing, continued to increase till 6 months (150% of control), and declined thereafter (125% at 9-10 months). In the contralateral soleus nerves the number of myelinated axons remained constant during this period. Nerve fibre diameters remained abnormally small; even after 10 months of reinnervation fibre diameters were unimodally distributed with a mean diameter of 3.3 microns in contrast to the bimodal distribution in intact nerves (mean values 3.9 and 9.0 microns, respectively). Total fibre cross-section area per nerve increased with time but reached only 54% +/- 6 S.D., (n = 3) of contralateral nerves by 10 months. The relative thickness of the myelin sheath (g-ratio) returned to normal after 9-10 months. Anatomically, muscle reinnervation appeared to be complete by 7-8 weeks since unusually small muscle fibre profiles were absent.(ABSTRACT TRUNCATED AT 400 WORDS)     

Effects of growth hormone on skeletal muscle. I. Studies on normal adult rats

A study was made on the effects of recombinant human growth hormone (rhGH) on fast and slow skeletal muscle in normal adult female rats. Daily injections of 4 IE of rhGH over 36 days resulted in increased levels of insulin-like growth factor I in serum and increased body weight. Morphometric analysis of the muscle fibres of the extensor digitorum longus (EDL) and soleus muscles revealed a significant increase in diameter of both type 1 and type 2 fibres in both muscles. The DNA: protein ratio and the number of satellite cells:muscle fibre in cross-sections was increased in the GH-treated rats in relation to controls. The results show that rhGH has pronounced effects on both cell proliferation and muscle fibre growth in skeletal muscle of normal adults rats.     

Late reinnervation of the rat soleus muscle is differentially suppressed by chronic stimulation and by ectopic innervation

Soleus (SOL) and extensor digitorum longus (EDL) muscles in adult rats were kept denervated for 2 months by four repeated freezes at 2-week intervals of the sciatic nerve. Reinnervation was studied in the absence or presence of chronic muscle stimulation, starting 1 month before reinnervation began. In addition, reinnervation was studied in SOL muscles where a previously transplanted fibular (FIB) nerve had formed ectopic neuromuscular junctions outside the original endplate area. After repeated freezes only, reinnervation was complete judged by tension measurements and histochemical examinations in SOL (n = 7) and EDL (n = 8) muscles. In directly stimulated muscles reinnervation was incomplete, and the force tensions evoked from indirect stimulation was on average 87 (n = 5) and 82% (n = 5) of direct muscle stimulation in SOL and EDL muscles, respectively. Of ectopically innervated SOL muscle fibres, only 26% became reinnervated in 12 muscles. Denervation and reinnervation increased the number of muscle fibres in stimulated (n = 4) and unstimulated (n = 5) EDL muscles by 18 and 15%, respectively. In stimulated (n = 4) and unstimulated (n = 7) SOL muscles, on the other hand, the number of muscle fibres remained normal. The stronger suppression of reinnervation in ectopically reinnervated compared to chronically stimulated SOL fibres indicates that reinnervation can also be suppressed by activity independent influences from the foreign nerve.     

Satellite cells and myonuclei in long-term denervated rat muscles

Background: The percentage of satellite cells rapidly decreases in aneurally regenerating soleus muscles of rat. Also denervation of intact muscles causes fiber loss and regeneration, but the fate of satellite cells is unknown; myonuclei have been suggested to undergo changes resembling those in apoptotic cells. Methods: Rat soleus and extensor digitorum longus (EDL) muscles were denervated at birth or at age 5 weeks and investigated after periods of up to 38 weeks. At least 400 myonuclei in each muscle were assessed by electron microscopy, and satellite cell nuclei were counted. In sity nick translation and tailing were performed after 30 weeks denervation in order to demonstrate DNA breaks associated with apoptosis. Results: Myotubes indicating regeneration were prominent in the adult denervated soleus and deep layers of EDL muscles after 7 weeks and in the superficial parts of EDL muscle after 16 weeks. The percentage of satellite cell nuclei slowly decreased to less than one fifth of normal after 20–30 weeks. Almost all satellite cells had vanished 10 weeks after neonatal denervation. Degenerating myonuclei in adult, but not in neonatally denervated muscles, remotely resembled apoptotic nuclei of lymphocytes, but no evidence of DNA breaks was found. Conclusion: Denervation of rat skeletal muscles causes, in addition to fiber atrophy, loss of fibers with subsequent regeneration. Proliferation of satellite cells under aneural conditions may lead to exhaustion of the satellite cell pool. This process is more rapid in growing than in adult muscles. Myonuclei in denervated muscles do not show DNA breaks which can be demonstrated by in situ nick translation. © 1995 Wiley-Liss, Inc.     

Paralysis of rat skeletal muscle equally affects contractile properties as does permanent denervation

The effects of long lasting (4--5 weeks) nerve conduction block and denervation were compared by investigating contractile, morphological and histochemical properties of slow (soleus) and fast (EDL) rat skeletal muscles. The block was based on improved perfusion techniques of the sciatic nerve with a tetrodotoxin (TTX) solution delivered at doses adequate to obtain maximal effects in the muscles. The TTX-inactivated axons retained normal histological and physiological properties such as the ability to evoke full contractile responses, to regenerate, and to completely reinnervate muscle. In spite of their intact innervation or of their full reinnervation, the TTX-paralysed muscles underwent weight loss, fibre atrophy and reduction in force output quantitatively indistinguishable from those following denervation. The same was true for all other contractile parameters tested, that is, twitch speed, twitch to tetanus ratio, post-tetanic potentiation, endurance, and fibre type composition. The results indicate the fundamental role of activity as a regulatory signal for muscle contractile properties, while they do not support the notion of a participation of chemical, activity-independent factors in this regulation This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of an anabolic hormone on striated muscle growth and performance

Chronic administration of an anabolic hormone, nandrolone phenylpropionate, in sedentary female rats for 6 weeks gave a 20% increase in body weight and the same proportional increase in all muscles sampled (heart, diaphragm, soleus, TA, EHP and EDL), such that the muscle/body weight ratio was unchanged. Cardiac muscle was unresponsive to treatment. Acute stimulation of EDL via lateral popliteal nerve gave similar values for contraction time, 1/2 relaxation time and twitch: tetanus ratio in both groups suggesting no slowing of the muscle. Fatigue resistance of EDL was improved with 0.29±0.029 vs. 0.46±0.071 of maximum isometric twitch tension being developed after 10 min repetitive stimulation at 4 Hz. This improved endurance was not accompanied by any increase in strength and could not be explained on the basis of cellular hypertrophy, but appears to reflect an increased aerobic capacity of skeletal muscle. The proportion of FOG fibres in EDL increased, 38±1.1% vs. 46±1.1%, and this was paralleled in the other skeletal muscles. Specific hypertrophy of FOG and FG fibres could be conclusively demonstrated in soleus and TA, respectively.     

The interaction between foreign and original motor nerves innervating the soleus muscle of rats.

1. The fibular nerve was transplanted on to the soleus muscle of the rats. Interruption of the original soleus nerve then permitted cross-innervation, and subsequently, over a period of weeks, re-innervation by the original nerve. 2. Individual muscle fibres were often innervated by both the original and the foreign nerve. The original and foreign end-plates were located in separate regions of the muscle. There were no indications that the original nerve could displace or repress the foreign innervation. 3. The extent of re-innervation by the original nerve depended upon the method of denervation. A single crush of the nerve was followed by virtually complete re-innervation, even of muscle fibres already innervated by the foreign nerve. When re-innervation was delayed by resection of a segment of the nerve only muscle fibres without foreign nerve innervation were re-innervated. Denervation by a simple nerve cut gave an intermediate result. 4. Re-innervation by the original nerve can take place without measurable extrajunctional sensitivity to ACh. 5. The original end-plate region could retain high and localized sensitivity to ACh for several months despite degeneration of its motor nerve terminal and activity of the muscle fibre. 6. Established foreign end-plates were re-innervated by the foreign nerve on muscle fibres with intact original innervation. 7. The factors controlling synapse formation in skeletal muscles are discussed.     

Effects of denervation and deafferentation on mass and enzyme activity in rat skeletal muscles

Effects of denervation and deafferentation on mass and enzyme activity of soleus and plantaris muscles were studied when the synergistic gastrocnemius was either intact or tenotomized in adult male Wistar rats. Denervation of the sciatic nerve at L4 and L5 induced a severe atrophy (approximately 50%) in both soleus and plantaris within 2 weeks. Deafferentation at L4 and L5 also caused a decrease in muscle weight for soleus and plantaris (approximately 11 and 24%, respectively). No compensatory response to the tenotomy was induced either for muscle weight or enzyme activity in both denervated and deafferentated muscles. It is suggested that the efferent information(s) through the nerve fibers has an important effect of the regulation of both muscle mass and enzyme activity. Its effect on the activities was greater in mitochondrial than cytoplasmic enzymes. The data also suggest that the afferent information(s) plays a significant role for the regulation of muscle mass, including an induction of compensatory hypertrophy, but not for enzyme activity.     

Muscle differentiation after sciatic nerve transection and reinnervation in adult rats

Reinnervation after peripheral nerve transections generally leads to poor functional recovery. In order to study whether changes in muscles might be a contributing factor in this phenomenon we studied muscle morphology and fibre type distributions after sciatic nerve transection in the rat hind limb. Proximally, before the bifurcation in the tibial and common peroneal nerve, a 12 mm segment of the sciatic nerve was resected, reversed and re-implanted as an autologous nerve graft. After survival periods of 7, 15 and 21 weeks the lateral gastrocnemius, tibialis anterior and soleus muscles were dissected, stained with mATP-ase, and fibre type distributions were studied. In addition, numbers of muscle fibres were counted, and cross sectional areas were calculated. After 7 weeks, cross sectional areas were decreased in all muscles. In the gastrocnemius and tibialis anterior muscles the fibre number remained unaltered but the hypotrophy had been reversed at later ages. The number of muscle fibres in the soleus muscle remained decreased over the entire period of observation. The percentages of type II fibres in the gastrocnemius and tibialis anterior muscles were decreased at 7 and 15 weeks but these again approached normal values at 21 weeks. The type I fibres, however, remained arranged in groups. In the soleus muscle a large increase in the percentage of type II muscle fibres was observed and this remained until 21 weeks. We conclude that a non-selective reinnervation and later readjustments by regression of polyneural innervation may in part explain the changes in distributions of various fibre types.     

A quantitative ultrastructural analysis of satellite cells in denervated fast and slow muscles of the mouse

Mononucleated cells located between the external lamina and sarcolemma of denervated muscle fibers within the extensor digitorum longus (EDL) and soleus muscles of adult mice were quantified and examined ultrastructurally from 3 to 65 days after ligating and removing a section of the sciatic nerve. During the first 2 weeks postdenervation, mononucleated cells in denervated muscles were morphologically indistinguishable from satellite cells observed in control muscles. With time, however, many of these satellite-like cells appeared more active as evidenced by a decrease in their nucleocytoplasmic ratio and an increase in their mean percentage of euchromatin material. The number of satellite cells (expressed as a ratio of satellite cell nuclei to satellite cell nuclei plus myonuclei) did not increase significantly until 30 days postdenervation, at which time the mean percentage for the soleus muscle had risen from a control value of 4.1–8.5%, and for the EDL from 1.2–4.1%. Smalldiameter, presumably regenerating, myofibers were occasionally observed but only after 30 days denervation. The ultrastructural evidence plus comparisons of euchromatin distributions between myonuclei and satellite cell nuclei support the concept that an increase in the number of satellite-like cells during denervation is more likely due to satellite cell proliferation than to the formation of mononucleated fragments utilizing preexisting myonuclei.     

'Fast' and 'slow' muscle fibres in hindlimb muscles of adult rats regenerate from intrinsically different satellite cells

Myosin heavy chain (MyHC) expression was examined in regenerating fast extensor digitorum longus (EDL) and slow soleus (SOL) muscles of adult rats. Myotoxic bupivacaine was injected into SOL and EDL and the muscles were either denervated or neuromuscularly blocked by tetrodotoxin (TTX) on the sciatic nerve. Three to 10 or 30 days later, denervated SOL or EDL, or innervated but neuromuscularly blocked EDL received a slow 20 Hz stimulus pattern through electrodes implanted on the muscles or along the fibular nerve to EDL below the TTX block. In addition, denervated SOL and EDL received a fast 100 Hz stimulus pattern. Denervated EDL and SOL stimulated with the same slow stimulus pattern expressed different amounts of type 1 MyHC protein (8% versus 35% at 10 days, 13% versus 87% at 30 days). Stimulated denervated and stimulated innervated (TTX blocked) EDL expressed the same amounts of type 1, 2A, 2X and 2B MyHC proteins. Cross-sections treated for in situ hybridization and immunocytochemistry showed expression of type 1 MyHC in all SOL fibres but only in some scattered single or smaller groups of fibres in EDL. The results suggest that muscle fibres regenerate from intrinsically different satellite cells in EDL and SOL and within EDL. However, induction by different extrinsic factors arising in extracellular matrix or from muscle position and usage in the limb has not been excluded. No evidence for nerve-derived trophic influences was obtained.     

Effects of chronic stimulation on the size and speed of long-term denervated and innervated rat fast and slow skeletal muscles

This study seeks to identify the mechanisms which motoneurones use to control the contractile force and speed of skeletal muscles. We have stimulated directly slow soleus (SOL) and fast extensor digitorum longus (EDL) muscles of adult rats intermittently at 100 Hz for 1-9 months. The muscles were either chronically denervated, denervated and reinnervated, or normally innervated. The stimulation started either immediately, or more commonly, after 1-9 months of denervation. Stimulation starting several months after denervation increased the mean maximum tetanic tension 37 times in SOL and eight times in EDL. These values represented 40 and 12% of the increases obtained by reinnervation after comparable periods of time. In denervated SOL and EDL muscles stimulated directly for more than 2 months, the mean isometric twitch contraction times were 13 and 12.7 ms, as in normal EDL muscles (13 ms). In innervated SOL muscles stimulated directly for 1-4 months, the mean twitch contraction times were 23.6 ms (normally innervated) and 19.2 ms (reinnervated), which were considerably shorter than in normal control SOL muscles (39.2 ms). Single motor unit recordings revealed that the natural (background) nerve impulse activity was essentially unaffected by the stimulation. Twitch contraction time and percentage of type II fibres in SOL muscles were related. The fastest muscles (denervated and stimulated) consisted of 100% type II fibres (with one exception), the second fastest (reinnervated and stimulated) of 70-50%, the third fastest (normally innervated and stimulated) of 45-0%, the second slowest (reinnervated) of 15-0%, and the slowest muscles (innervated controls) of 5-0% type II fibres.     

Myogenic and neurogenic regulation of myosin gene expression in cat jaw-closing muscles regenerating in fast and slow limb muscle beds

Summary Immunocytochemical techniques were used to study changes in myosin gene expression during the regeneration of the cat posterior temporalis muscle transplanted into the bed of either the fast extensor digitorum longus (EDL) or the slow soleus muscle. Strips of the posterior temporalis, a homogeneously superfast muscle, were treated with Marcaine and then transplanted into limb muscle beds which had been completely cleared of host muscle fibres. The regenerates were examined 6 to 224 days after surgery. Early regenerates in both muscle beds reacted with antibodies against the heavy chain of foetal, slow and superfast myosins, but not with antibodies against fast myosin. In the long-term, regenerates innervated by the EDL nerve expressed only superfast myosin whereas in the regenerates innervated by the soleus nerve most fibres expressed only slow myosin and only a few fibres reacted exclusively with the anti-superfast myosin antibody even after 210 days. In contrast, EDL and soleus muscles regenerating in their own beds expressed foetal, slow and fast myosin, but did not express superfast myosin. The isometric contraction times of the various types of regenerates reflected the types of myosin synthesized. It is concluded that jaw and limb muscle cells exist as two distinct allotypes, each having a distinct repertoire for the expression of adult isomyosins, and that within that repertoire isomyosin gene expression can be modulated by the nerve. Thus, myosin gene expression in skeletal muscle fibres is regulated by both myogenic and neurogenic mechanisms.     

Increase of elastic fibres in muscle spindles of rats following single or repeated denervation with or without reinnervation

Summary Muscle spindles in the lower lumbrical muscles of rats were studied by transmission electron microscopy following denervation with or without reinnervation. The number and total area of elastic fibres per muscle spindle increased at 3–12 months following various experimental procedures: (1) denervation and reinnervation after a single crush lesion to the sciatic nerve; (2) reinnervation after four-fold repeated crush injuries; and (3) transection and suture of the nerve. The increased number of oxytalan and elaunin fibres, the precursors of mature elastic fibres, within these muscle spindles provided further evidence for their numerical and dimensional increase. An attachment site of elastic fibres at the spindle pole was identified at the inner cells of the outer spindle capsule. The processes of these cells embraced terminating elastic fibres tightly. Attachment of elastic fibres to intrafusal muscle fibres was less conspicuous since they were not similarly embraced but were rather indistinctly, though closely, associated with the basal lamina along longitudinal surface indentations of intrafusal muscle fibres. It is concluded from this series of experiments that muscle spindles, as dynamic mechanoreceptors, maintain their elastic properties even under pathological conditions. The increase of elastic fibres following denervation and reinnervation represents an obviously meaningful reaction that may compensate for loss of tonic properties of muscle spindles without causing stiffness.     

Nestin expression reflects formation, revascularization and reinnervation of new myofibers in regenerating rat hind limb skeletal muscles

This work provides a comprehensive view of expression and distribution patterns of the intermediate filament nestin in rat regenerating skeletal muscles. Regeneration was induced by heterochronous isotransplantation of extensor digitorum longus (EDL) or soleus muscles from 15-day-old rats into the EDL muscle of adult female inbred Lewis rats. Recipients were sacrificed by anesthetic overdose and the host muscles with the graft were excised after 7-, 16-, 21- and 29-day survival, fixed in 10% formalin solution and immunohistochemically stained. In the regenerating skeletal muscle maximal nestin expression was observed in newly formed myoblasts and myotubes and it decreased in the course of skeletal myofiber development. We demonstrate the phenomenon of a peripheral concentration of nestin expression within the differentiating myofibers called 'lateralization', which may represent a mechanism of nestin elimination during the development of skeletal myofibers. Furthermore, nestin has been detected in newly formed endothelial cells of blood vessels growing into the regenerated area and also in peripheral nerves innervating the graft as well as the host muscle. In conclusion, according to our results, nestin represents an ideal marker for skeletal muscle regeneration, as its expression reflects processes of development, revascularization and reinnervation of new myofibers.     

The use of local anaesthetics to produce prolonged motor nerve block in the study of denervation hypersensitivity

1. Silastoseal cuffs containing 25% and 45% lignocain, 25% marcain and 25% NaCl were placed onto the sciatic nerve. The effects of this on the sciatic nerve as well as the soleus and extensor digitorum longus (EDL) muscles was studied. 2. Cuffs containing local anaesthetics caused both early and late phases of paralysis, cuffs containing NaCl caused only the late phase of paralysis, showing that the late phase of paralysis was not due to the anaesthetic properties of the drugs. The paralysis produced by these procedures was irreversible. 3. Responses to indirect and direct stimulation of soleus and EDL muscles were compared to assess the degree of denervation. In case of cuffs containing 25% marcain and 45% lignocain more than half of the muscle fibres were denervated. 4. Acetylcholine sensitivity was assesses in all experiments. Soleus and EDL muscles from all animals that had cuffed nerves containing either local anaesthetics or 25% NaCl were hypersensitive to ACh. The degree of hypersensitivity could be correlated to the degree of "denervation" as assessed by comparing the directly and indirectly elicited twitch tensions. 5. It was found that the sciatic nerve undergoes degenerative changes when exposed to cuffs containing either local anaesthetics or 25% NaCl.