Peripheral nerve injury and recovery after temporary ischemia

Summary Nerve (NCV) and motor (MNCV) conduction velocities of the rat sciatic nerve were examined between 1 and 90 days after ischemia for 1, 2, 3, 4 or 6 h. The results were compared to light and electron microscopy of the nerve.Slight diminution in the MNCV was observed 1 day after 1–2 h ischemia, whereas 3–6 h ischemia resulted in a complete conduction block. Diminution in the NCV occurred first after ischemia for 2 h and a complete block was seen after 4 and 6 h ischemia. Reduced NCV and MNCV were seen between 4 and 18 days only in the animals subjected to ischemia of longer duration of 3–6 h. Both the NCV and MNCV were nearly normalized at the 90th day. Ischemia of 4 and 6 h resulted in denervation of some of the muscle fibers, seen as spontaneous fibrillation at the 4th and 18th day.Electron microscopy and histometric studies showed degeneration of myelinated fibers increasingly after longer durations of ischemia; ischemia for 2 h caused a degeneration of about 5%, 3 h of about 35%, 4 h about 45%, and 6 h about 75% of the fibers. Myelinated fibers of different sizes were equally damaged. In the teased fiber preparations normal and myelin sheaths undergoing Wallerian-like degeneration was seen. Regeneration occurred, but even at the 90th day there was a tendency of the myelin/axon ratio towards values less than control values.     

Quantitation of axon loss and conduction block in acute radial nerve palsies.

Eleven acute radial nerve palsies were examined between 3 days and 14 weeks following the onset of the neuropathy. Our objective was to quantify the relative extent of axon loss and conduction block in radial motor fibers supplying the extensor and abductor pollicis longus (EPL/APL) muscles. In 10 of 11 cases, conduction block exceeded axon loss. Maximum motor and sensory conduction velocities were normal distal to the spiral groove, suggesting that the larger myelinated fibers were not selectively involved in this acute neuropathy. The localization of the conduction block and slowing was, in every case, across the spiral groove. This method provides a relatively simple way of assessing the approximate contributions of axon loss and conduction block, and fits well with the early and usually complete clinical recovery in these cases.     

Rapid alterations of the axon membrane in antibody-mediated demyelination

Alterations of nodal and paranodal axolemma of the rat sciatic nerve were investigated in antigalactocerebroside serum-induced demyelination. A ferric ion-ferrocyanide (FeFCN) stain that appears to stain the regions with a high sodium channel density in nerve fibers was applied. When acute conduction block was initiated 20 to 180 minutes after the antiserum injection, myelin terminal loops began to be detached from the paranodal axolemma and reaction product of FeFCN stain originally localized at the nodes decreased in density and extended to the paranodal axolemma. By the time that complete conduction block was established, 5 hours after the injection, FeFCN stain was barely detectable around the nodal area. The loss of staining was associated with detachment and vesiculovacuolar degeneration of the paranodal myelin. This rapid deterioration and disappearance of normal cytochemical characteristics of the axolemma in the presence of only modest paranodal demyelination could be a morphological correlate of the loss of excitability of the axon membrane.     

Physiopathology of the clinical manifestation of multiple sclerosis. Electrophysiological hypotheses

Clinical manifestations of multiple sclerosis cannot always be correlated with the type of lesion present. Plaques in certain regions may be clinically silent whereas, inversely, some neurological disorders appear not to be related to a demyelinization process. The pathophysiology of neurological symptoms and signs in multiple sclerosis may therefore be related to two other factors: 1) membrane phenomena observed in demyelinated fibers and leading to partial or complete conduction block, or 2) conduction blocks due to pathological phenomena that fail to provoke anatomical demyelinization. Demyelinization induces a sudden drop in impedance of the axon membrane which is responsible for the conduction block. Redistribution of sodium ionophores in the demyelinated axon enables transmission of certain messages but in a slower and unreliable manner. Sensitivity of demyelinated fibers to temperature and the extracellular ionic environment provides an explanation for the transient, recurrent stereotyped reactions provoked by fever, physical exercise or digestion. In the absence of demyelinization, a conduction block could be the result of either a minimal lesion at the paranodal myelin with denudation of the specialized parts between axon and glia,or of humoral blocking factors--of debatable specificity--or cellular factors that have been demonstrated recently. The existence of such mechanisms suggests the need for revision of the notion of acute episodes of multiple sclerosis. Finally, knowledge of the mechanisms involved in nerve conduction along demyelinated fibers suggests the possibility of therapy to restore conduction of these fibers by acting on extracellular ionic concentrations or directly on membrane ionic canals.     

Ganglioside promotes the bridging of sciatic nerve defects in cryopreserved peripheral nerve allografts

Previous studies have shown that exogenous gangliosides promote nervous system regeneration and synapse formation.In this study,10 mm sciatic nerve segments from New Zealand rabbits were thawed from cryopreservation and were used for the repair of left sciatic nerve defects through allograft bridging.Three days later,1 m L ganglioside solution(1 g/L) was subcutaneously injected into the right hind leg of rabbits.Compared with non-injected rats,muscle wet weight ratio was increased at 2–12 weeks after modeling.The quantity of myelinated fibers in regenerated sciatic nerve,myelin thickness and fiber diameter were elevated at 4–12 weeks after modeling.Sciatic nerve potential amplitude and conduction velocity were raised at 8 and 12 weeks,while conduction latencies were decreased at 12 weeks.Experimental findings indicate that ganglioside can promote the regeneration of sciatic nerve defects after repair with cryopreserved peripheral nerve allografts.     

Morphologic basis of conduction block]

Conduction block is electrophysiologically defined and is shown to be an important pathologic condition encountered in both central and peripheral nervous system disorders. The conduction block is encountered most frequently in peripheral nerve injuries, which are the results of compression and ischemia. It is impossible to study morphologically the particular myelinated fibers showing the conduction block in human cases, but the alterations of myelinated fibers in the experimental models have been studied. The disturbance and/or destruction of the axoglial junction, with or without subsequent paranodal and segmental demyelination, is the major cause of the conduction block and is relatively easily identified morphologically in teased fiber and electron microscopic preparations of myelinated fibers. Following paranodal and segmental demyelination, the compensatory increase of large intramembranous particles, probably corresponding to the sodium channel, may enable the continuous conduction to be successful across the demyelinated axon. The abnormality of the nodal axolemma, dysfunction and/or loss of sodium channels, is another major cause and is not easily proven by light and electron microscopic techniques. Both causes may be concomitant under certain clinical and experimental conditions. Clinical and experimental conditions characterized by the conduction block are also listed.     

Focal conduction block in the dorsal root ganglion in experimental allergic neuritis.

Acute experimental allergic neuritis was induced in Lewis rats by inoculation with bovine intradural root myelin and adjuvants. In terminal experiments, sensory conduction was assessed in rats with hindlimb ataxia and weakness by stimulating the exposed sciatic nerve and recording directly from the exposed L-4 spinal nerve, dorsal root ganglion, dorsal root, and dorsal root entry zone. Focal conduction block was present in a high proportion of large-diameter fibers in the dorsal root ganglion. In contrast, nerve conduction in the peripheral nerve and spinal nerve was essentially normal apart from probable conduction block in some fibers in the proximal spinal nerve in a minority of rats. The afferent volley arriving at the dorsal root entry zone of the spinal cord was greatly reduced, as a consequence of the conduction block in the dorsal root ganglion and probable conduction block in the dorsal root. The M wave recorded from the fourth dorsal interosseus muscle of the hindfoot was normal in amplitude but slightly prolonged in latency and the H reflex was absent. These electrophysiological findings correlated well with the histological findings of inflammation and prominent demyelination in the dorsal root ganglia and dorsal roots with minimal involvement of the proximal spinal nerve and no involvement of the sciatic nerve. It is concluded that the hindlimb ataxia in rats with this form of acute experimental allergic neuritis is due to demyelination-induced nerve conduction block in the dorsal root ganglia and probably in the dorsal roots.     

Effects of tacrolimus (FK506) on sciatic nerve regeneration in rats

BACKGROUND: Tacrolimus (FK506) protects peripheral nerves located in damaged regions by inhibiting T lymphocyte proliferation and activation.OBJECTIVE: To evaluate the effect of FK506 on promoting regeneration of rat sciatic nerve. DESIGN, TIME AND SETTING: A randomized, controlled, animal study was performed at the Laboratory of the Department of Orthopedic Surgery, Dalian Medical University, China, from September 2007 to September 2008.MATERIALS: A total of 60 adult, male, Sprague-Dawley rats were equally and randomly divided into model, local administration and systemic administration groups. All rats received a neurotomy of bilateral sciatic nerves to establish models of nerve regeneration chambers. The powder and injection of FK506 were supplied by Fujisawa Pharmaceutical, Japan.METHODS: The regeneration chambers of the model group were infused with 0.2 mL saline. The systemic group were injected with 0.2 mL saline, followed by daily subcutaneous injections of FK506 (1 mg/kg), for 14 days. The local administration group was infused with 0.2 mL FK506 (1 μg/mL).MAIN OUTCOME MEASURES: Local immune response was observed using hematoxylin-eosin staining. Myelinated nerve fiber number, myelin sheath and nerve fiber thickness were observed using toluidine blue staining. Wet weight of gastrocnemius was evaluated. Compound muscle action potential amplitude, latency, and conduction time were recorded, and motor nerve conduction velocity was calculated using electrophysiology.RESULTS: The total number of myeiinated nerve fibers in the local and systemic administration groups was significantly higher than in the model group. The density of myelinated nerve fibers, myelin sheath thickness and mean axon diameter were significantly increased in the systemic administration group compared with the model group (P < 0.05). Lymphocyte infiltration was decreased in the local and systemic administration groups compared with the model group. The wet weight of rat gastrocnemius in the local and systemic administration groups were significantly greater compared with the model group (P<0.05). Motor nerve conduction velocity was the fastest in the systemic administration group, and the slowest in the model group. Compound muscle action potential amplitude was larger in the systemic administration group compared with the local administration and model groups (P<0.05).CONCLUSION: Systemic administration of FK506 can promote regeneration of rat sciatic nerve and recovery of neural function. Systemic administration produced better regeneration and recovery of function than local administration of FK506.     

Acrylamide-induced peripheral neuropathy in normal and neurofilament-deficient Japanese quails

Morphological effects of acrylamide (AC) on the peripheral nerves in normal and neurofilament (NF)-deficient (Quv) Japanese quails were investigated. AC (100 mg/kg) was injected intraperitoneally every other day. After the birds manifested neurological signs, they were necropsied (after 10 21 AC injections) and the sciatic and tibial nerves were examined. In both normal and Quv qualis, AC produced axonopathy with a distal-proximal progression. In AC-intoxicated normal quails, the nerve fiber pathology was characterized by typical Wallerian-like degeneration, consisting of axonal degeneration, myelin breakdown, macrophage migration, Schwann cell proliferation and regeneration of nerve fibers. Ultrastructurally, AC-induced NF accumulation was detected in the axon of myelinated nerve fibers. In AC-intoxicated Quv qualis, axonal degeneration with accumulation of membranous organelles occurred; however, sequential events of Wallerian-like degeneration were not as prominent as in AC-intoxicated normal qualis. These results demonstrated that NF-deficient Quv quails are sensitive to neurotoxic effects of AC. On the other hand, the different pathology of AC-intoxicated normal and Quv qualis indicates the presence or absence of NFs influences the appearance and extent of AC axonopathy.     

Conduction studies in peripheral cat nerve using implanted electrodes: III. The effects of prolonged constriction on the distal nerve segment

Electrophysiological properties were monitored in detail in chronically constricted peripheral nerves by implanted, multicontact nerve cuff electrodes and correlated with morphometric histology in selected cases. The physiological and histological responses in nerve to a range of constricting cuffs of standard sizes were readily graded. The initial response to any significant constriction was a transient, focal conduction slowing or block at the constriction, followed by more protracted distal effects; the latter ranged from loss of excitability consistent with "dying-back" degeneration to reductions in conduction velocity consistent with histologically observed atrophy. Smaller myelinated fibers tended to have similar but less pronounced changes than larger diameter fibers. Recordings from ventral and dorsal roots showed that distal degeneration was more pronounced in motor than in sensory fibers of similar caliber. Electronmicroscopical measurements showed that basal laminas were relatively preserved around even the most atrophic and demyelinated axons. Perimeter measurements of the basal lamina could be used to estimate the diameter of the original nerve fiber.     

Remyelination and recovery of conduction in cat optic nerve after demyelination by pressure

Pressure has been applied to the optic nerve of cats sufficient to block conduction in the large (Y) nerve fibers. The pressure block produces a mixture of axotomy and demyelination. By means of implanted electrodes, recovery of conduction in these fibers was monitored. There is a short-term recovery starting about 2 weeks after block induction and finishing at about 4 weeks. A later recovery starts at about 6-7 weeks and finishes at about 10-11 weeks. The remyelination has been monitored in the electron microscope by measurement of the myelin thickness and axon diameter of the large fibers. The remyelination follows a time course similar to the late phase of conduction recovery. By reference to the work of others, we surmise that the early recovery of conduction is due to the reorganization of microtubules disorganized by the pressure.     

Recovery of brain dysfunction after methylmercury exposure in rats

We studied the time course of central nervous system (CNS) involvement after the termination of methylmercury exposure to rats, in order to investigate whether or not the involvement still progresses even after the termination of exposure. Methylmercury chloride (MMC), at a dose of 2 mg/kg/day, was subcutaneously injected for 25 consecutive days in 12 adult male Sprague-Dawley rats. Six of them were sacrificed on the final day of exposure (group A) after completing the observations of behavioral changes and determining the local cerebral glucose utilization (LCGU) as an indicator of cerebral neuronal activities. Histological examinations of the brain and the sciatic nerve were done. The other six rats were further followed up for 90 days after the termination of exposure (group B). In addition, six rats that received physiological saline served as a control. Group A showed a significant reduction of LCGU without any accompanying cerebral histological alterations and a moderate loss of myelinated fibers in the sciatic nerve. Group B showed normal LCGU rates while severe axonal degeneration of the sciatic nerve was found on the final day of the 90-day follow-up period. The present results demonstrate that a transient involvement of the CNS can occur after MMC exposure. In addition, a complete recovery may occur when the process is mild enough not to cause histological alterations. In contrast, the involvement of the peripheral nerve is much more severe than that of the CNS and it was observed to progress even after the cessation of MMC exposure. Therefore, it seems unlikely, at least in rats, that a steadily progressive course occurs in the CNS but not in the peripheral nerves over a long period of time after MMC exposure.     

The effect of mixed bovine brain gangliosides on hypoxic conduction block in control and streptozotocin-diabetic rats.

This study describes the electrophysiological responses of endoneurial preparations derived from rat sciatic nerve to acute hypoxia in vitro. Preparations from control rats exhibited a 40% decline in compound action potential (CAP) amplitude after 40 min exposure to medium gassed with 8% O2. In preparations from 4 week streptozotocin-diabetic rats CAP declined by only 29%, indicating a resistance to hypoxic conduction blockade. Treating diabetic rats with mixed bovine brain gangliosides (10 mg/kg/day i.p.) exaggerated this resistance to hypoxic conduction blockade as CAP amplitude fell to only 18% of initial values. In a separate experiment, treating non-diabetic rats with gangliosides (10 mg/kg/day i.p.) or adding gangliosides (400 micrograms/ml) directly to the medium in which control nerves were maintained during in vitro recording also significantly attenuated the decline in CAP amplitude after 40 min hypoxia, thus effectively inducing a resistance to hypoxic conduction blockade similar to that observed in nerves from diabetic rats. These studies demonstrate that the systemic or acute local administration of gangliosides induces a resistance to hypoxic conduction block in normal nerve and exaggerates the resistance to hypoxic conduction block of diabetic rats.     

Nerve conduction changes and fine structural alterations of extra- and intrafusal muscle and nerve fibers in streptozotocin diabetic rats

Streptozotocin-induced diabetes mellitus in known to cause a reduction of both conduction velocity and axon caliber in sciatic nerves and also a decrease in muscle fiber size. The present study investigates whether the distal parts of the peripheral nervous system, including extra- and intrafusal muscle fibers, are more severely affected than the proximal segments in the diabetic state. Proximal and distal sensory nerve conduction velocities were monitored during a period of 3 months in rats rendered diabetic by injection of streptozotocin. Segments of the sciatic and ventral coccygeal nerves, and of the biceps femoris and lumbrical muscles, were studied by light and electron microscopy, including morphometric analysis. In contrast to previous studies, daily suboptimal insulin injections were given to prevent acute metabolic complications. Sensory conduction velocity in the ventral coccygeal nerve was significantly (P > 0.05) decreased in the diabetic rats compared to controls. Proximal and distal nerve segments were equally affected. Mean cross-sectional axon area of the sciatic nerve was moderately, but significantly (P < 0.05), smaller in insulin-treated diabetic rats than in controls. In both the sciatic nerve and the terminal, intrafusal nerve segments, occasional axons showed moderate dystrophic changes. Fibers of the intrafusal nerve segments appeared to be equally affected compared to the fibers in the sciatic nerve, although no quantitative comparison was made. The increase of small caliber skeletal muscle fibers in experimental streptozotocin-induced diabetes was confirmed. These findings indicate that proximal and distal segments of peripheral nerves are affected equally in the early stages of experimental diabetic neuropathy.© 1995 John Wiley &Sons, Inc.     

Recombinant human fibroblast growth factor-2 promotes nerve regeneration and functional recovery after mental nerve crush injury

Several studies have shown that fibroblast growth factor-2(FGF2) can directly affect axon regeneration after peripheral nerve damage. In this study, we performed sensory tests and histological analyses to study the effect of recombinant human FGF-2(rh FGF2) treatment on damaged mental nerves. The mental nerves of 6-week-old male Sprague-Dawley rats were crush-injured for 1 minute and then treated with 10 or 50 μg/m L rh FGF2 or PBS in crush injury area with a mini Osmotic pump. Sensory test using von Frey filaments at 1 week revealed the presence of sensory degeneration based on decreased gap score and increased difference score. However, at 2 weeks, the gap score and difference score were significantly rebounded in the mental nerve crush group treated with 10 μg/m L rh FGF2. Interestingly, treatment with 10 μg/m L rh FGF had a more obviously positive effect on the gap score than treatment with 50 μg/m L rh FGF2. In addition, retrograde neuronal tracing with Dil revealed a significant increase in nerve regeneration in the trigeminal ganglion at 2 and 4 weeks in the rh FGF2 groups(10 μg/m L and 50 μg/m L) than in the PBS group. The 10 μg/m L rh FGF2 group also showed an obviously robust regeneration in axon density in the mental nerve at 4 weeks. Our results demonstrate that 10 μg/m L rh FGF induces mental nerve regeneration and sensory recovery after mental nerve crush injury.     

Effects of dexmedetomidine and dexketoprofen on the conduction block of rat sciatic nerve

Dexmedetomidine is a selective α2-adrenoceptor agonist that is used because of its sedative,anxiolytic,and analgesic effects.Dexketoprofen,which is used as an analgesic,is a nonselective nonsteroidal anti-inflammatory drug (NSAID).The use of dexmedetomidine and dexketoprofen as adjuvants to local anesthetics for the peripheral nerve is gradually increasing.In this study,we aimed to investigate the effects of different doses of dexmedetomidine and dexketoprofen on conduction block of rat sciatic nerve.The isolated sciatic nerve from adult rats was transferred to a nerve chamber.The compound action potentials (CAPs) were recorded from stimulated nerve with electrophysiological methods.Dexmedetomidine (n = 8) and dexketoprofen (n = 8) were administered in the chamber with cumulative concentrations of 10–9 to 10–5 M,and the CAPs were recorded for 5 and 10 minutes.The CAP parameters were calculated.Both dexmedetomidine and dexketoprofen significantly depressed all CAP parameters in a dose-dependent manner compared with the control group,i.e.,the group in which rats did not receive treatment.CAP parameters showed there was no significant difference in nerve conduction inhibition between dexmedetomidine and dexketoprofen.Higher doses of dexmedetomidine suppressed the conduction in the fast-conducting fibers;however,dexketoprofen was found to suppress the conduction in the slow-conducting fibers in a time-dependent manner and suppress the conduction in the medium- and slow-conducting fibers in a dose-dependent manner.These findings suggest that dexmedetomidine and dexketoprofen exhibit better anesthetic effects on peripheral nerve through different ways of action.The experimental procedures were approved by the Necmettin Erbakan University on January 30,2013 (approval No.2013-024).     

Chronic inflammatory demyelinating polyneuropathy--an electrophysiological and histological study of the ulnar nerve from a case with intractable persistent conduction block]

We report a 71-year-old male with CIDP, in whom complete motor conduction block persisted for more than several years. Corticosteroid and plasma therapy showed little effect to his weakness. He died of pneumonia. The ulnar nerve in which complete conduction block had persisted was taken out immediately after death and studied with conventional histological and morphometrical techniques. In the transverse section, many thinly-myelinated fibers and some cluster formations were confirmed. Active axonal degeneration was scarce. Myelinated fiber density was 7,417/sgmm. Fiber diameter histogram showed a two-peaked normal distribution. It is thus concluded that demyelination with little axonal degeneration can be the main pathological process causing intractable weakness and poor prognosis of CIDP. Low CMAPs should not lead to therapeutic nihilism, because it may simply be caused by demyelination without exonal degeneration in CIDP.     

A mild acute compression induces neurapraxia in rat sciatic nerve

The pressure that induces neurapraxia in rat remains unrevealed. To determine the appropriate force to induce neurapraxia, two types of clips were applied to the sciatic nerve and were evaluated with functional, electrophysiological, and histological examinations. With a compression of 60 g/mm2, walking track analysis showed complete sciatic nerve paralysis one day postoperatively, but became normal in 14 days. Electrophysiologically, complete conduction block occurred one day post operatively, whereas the motor conduction velocity (MCV) below the compression site remained normal. Histologically, only limited signs of Wallerian degeneration were seen. The model in this study exhibited the features of neurapraxia.     

A MILD ACUTE COMPRESSION INDUCES NEURAPRAXIA IN RAT SCIATIC NERVE

The pressure that induces neurapraxia in rat remains unrevealed. To determine the appropriate force to induce neurapraxia, two types of clips were applied to the sciatic nerve and were evaluated with functional, electrophysiological, and histological examinations. With a compression of 60 g/mm2, walking track analysis showed complete sciatic nerve paralysis one day postoperatively, but became normal in 14 days. Electrophysiologically, complete conduction block occurred one day post operatively, whereas the motor conduction velocity (MCV) below the compression site remained normal. Histologically, only limited signs of Wallerian degeneration were seen. The model in this study exhibited the features of neurapraxia.     

Alterations in the structure, function, and chemistry of C fibers following local application of vinblastine to the sciatic nerve of the rat

Vinblastine, a transport blocker, was applied locally to the sciatic nerve in rats. It was found to be a powerful neurotoxin with a dose-dependent action, destroying all afferents at doses of 5 X 10(-4)M, primarily C fibers at intermediate doses of 2.5 X 10(-4)M, and only at a critically low dose of 10(-4)M was a degeneration-free axon transport blockade, lasting for 4 to 5 days, produced. Such transport block failed to alter thermal responsiveness of the rats as measured behaviorally, by the flexor reflex, or by dorsal horn cell responses. It did, however, significantly reduce both the chemical sensitivity of the C afferents and their ability to produce neurogenic edema. This began 24 hr after treatment and lasted 4 to 5 days. Therefore, it is likely that these functions are dependent on the continuous transport of some compound to the axon terminals from the cell body. This low concentration of local vinblastine treatment also resulted in depletion of fluoride-resistant acid phosphatase from C fiber terminals in the dorsal horn of the spinal cord. Transmission from C fibers to second-order neurons in the spinal cord, however, was totally unaffected. Substance P levels in the spinal terminals was largely unaffected, although in 1 of 5 cases there was depletion. It appears, therefore, that some, but not all, retrograde changes in sensory neurons following peripheral nerve damage can be mimicked by blockade of axon transport. The effects following vinblastine treatment are compared to other peripheral nerve manipulations, such as cut, crush, and application of local capsaicin.