Claudin 14/15 play important roles in early wallerian degeneration after rat sciatic nerve injury / 中华创伤杂志(英文版)

PURPOSE@#Wallerian degeneration (WD) is an antegrade degenerative process distal to peripheral nerve injury. Numerous genes are differentially regulated in response to the process. However, the underlying mechanism is unclear, especially the early response. We aimed at investigating the effects of sciatic nerve injury on WD via CLDN 14/15 interactions in vivo and in vitro.@*METHODS@#Using the methods of molecular biology and bioinformatics analysis, we investigated the molecular mechanism by which claudin 14/15 participate in WD. Our previous study showed that claudins 14 and 15 trigger the early signal flow and pathway in damaged sciatic nerves. Here, we report the effects of the interaction between claudin 14 and claudin 15 on nerve degeneration and regeneration during early WD.@*RESULTS@#It was found that claudin 14/15 were upregulated in the sciatic nerve in WD. Claudin 14/15 promoted Schwann cell proliferation, migration and anti-apoptosis in vitro. PKCα, NT3, NF2, and bFGF were significantly upregulated in transfected Schwann cells. Moreover, the expression levels of the β-catenin, p-AKT/AKT, p-c-jun/c-jun, and p-ERK/ERK signaling pathways were also significantly altered.@*CONCLUSION@#Claudin 14/15 affect Schwann cell proliferation, migration, and anti-apoptosis via the β-catenin, p-AKT/AKT, p-c-jun/c-jun, and p-ERK/ERK pathways in vitro and in vivo. The results of this study may help elucidate the molecular mechanisms of the tight junction signaling pathway underlying peripheral nerve degeneration.

Clinical and Radiological Features of Wallerian Degeneration of the Middle Cerebellar Peduncles Secondary to Pontine Infarction / 中华医学杂志(英文版)

<p><b>Background</b>Wallerian degeneration (WD) of bilateral middle cerebellar peduncles (MCPs) can occur following pontine infarction, but its characteristics have not yet been clarified because of the low incidence. Thus, the present study discussed the clinical and radiological features to improve the awareness of this disease.</p><p><b>Methods</b>Clinical and radiological information from consecutive individuals diagnosed with WD of bilateral MCPs following pontine infarction in three hospitals over the past 4 years between October 2012 and October 2016 were retrospectively investigated and compared with a control group (patients with pontine infarction had no secondary WD).</p><p><b>Results:</b>This study involved 30 patients with WD of MCPs, with a detection rate of only 4.9%. The primary infarctions (χ =24.791, P = 0.001, vs. control group) were located in the paramedian pons in 21 cases (70.0%), and ventrolateral pons in nine cases (30.0%). WD of the MCPs was detected 8-24 weeks after pons infarction using conventional magnetic resonance imaging (MRI); all secondary WDs were asymptomatic and detected incidentally. All WD lesions exhibited bilateral, symmetrical, and boundary blurring on MRI. The signal features were hypointense on T1-weighted imaging, hyperintense on T2-weighted imaging and fluid-attenuated inversion recovery, and slightly hyperintense or isointense on diffusion-weighted imaging and apparent diffusion coefficient maps. Secondary brainstem atrophy was found in six (20.0%) cases. A Modified Rankin Scale score 0-2 was found in 10 (33.3%) cases and score >2 in 20 (66.7%) cases at 90 days after discharge, and the short-term prognosis was worse than that in control group (χ =12.814, P = 0.001).</p><p><b>Conclusions</b>Despite the rarity of bilateral and symmetrical lesions of MCPs, secondary WD should be highly suspected if these lesions occur within 6 months after pontine infarction, particularly paramedian pons. Conventional MRI appears to be a relatively sensitive method for detecting WD of MCPs, which might affect the short-term prognosis.</p>

Célula de Büngner / Büngner cell

Las lesiones del nervio periférico constituyen una condición clínica frecuente; por ello, entender su fisiopatología y los avances en el campo de la regeneración nerviosa es fundamental para brindar el mejor tratamiento a los pacientes. En los últimos años se ha venido dando cada vez mayor importancia a los eventos regenerativos después de la lesión, donde interviene en gran medida una expresión fenotípica única en este proceso, derivada de células ya presentes, fenómeno clave para la recuperación de la función del nervio lesionado. Este artículo revisó la literatura disponible con el objetivo de entender mejor este evento regenerativo y se encontraron procesos celulares y moleculares que suceden en los axones.

Peripheral nerve injuries are a common clinical condición for which the understanding of the pathophysiology and advances in the fteld of nerve regeneration are important to provide the best treatment for patients. In recent years, it has been giving increasing importance to the regenerative events after injury, where it operares largely unique phenotypic expression in this process, derived from cells already present, kev event for the recoven- of nerve function injured. A review of the literature is done with the aim of a better understanding of this regenerative event, fínding a series of cellular and molecular processes that go on axonal level.

Current State and Prospects of Development of Blood-based Biomarkers for Mild Traumatic Brain Injury

The current understanding of the pathophysiology of mild traumatic brain injury (mTBI) is, without doubt, incomplete. Nevertheless, we tried to summarize the state-of-the-art explanation of how the brain is continuously injured even after a single impact. We also reviewed the real struggle of diagnosing mTBI, which culminated in showing the potential of blood-based biomarkers as an alternative or complementary way to overcome this difficulty. Pathophysiology of mTBI is subdivided into primary and secondary injuries. Primary injury is caused by a direct impact on the head and brain. Secondary injury refers to the changes in energy metabolism and protein synthesis/degradation resulting from the biochemical cascades as follows; calcium influx, mitochondrial dysfunction, fractured microtubules, and Wallerian degeneration, neuroinflammation, and toxic proteinopathy. Since the diagnosis of mTBI is made through the initial clinical information, it is difficult and inaccurate to diagnose mTBI without the absence of a witness or sign of head trauma. Blood-based biomarkers are expected to play an important role in diagnosing mTBI and predicting functional outcomes, due to their feasibility and the recent progress of targeted proteomics techniques (i.e., liquid chromatography tandem mass spectrometry [LC-MS/MS]).

Rescuing axons from degeneration does not affect retinal ganglion cell death

After a traumatic injury to the central nervous system, the distal stumps of axons undergo Wallerian degeneration (WD), an event that comprises cytoskeleton and myelin breakdown, astrocytic gliosis, and overexpression of proteins that inhibit axonal regrowth. By contrast, injured neuronal cell bodies show features characteristic of attempts to initiate the regenerative process of elongating their axons. The main molecular event that leads to WD is an increase in the intracellular calcium concentration, which activates calpains, calcium-dependent proteases that degrade cytoskeleton proteins. The aim of our study was to investigate whether preventing axonal degeneration would impact the survival of retinal ganglion cells (RGCs) after crushing the optic nerve. We observed that male Wistar rats (weighing 200-400 g; n=18) treated with an exogenous calpain inhibitor (20 mM) administered via direct application of the inhibitor embedded within the copolymer resin Evlax immediately following optic nerve crush showed a delay in the onset of WD. This delayed onset was characterized by a decrease in the number of degenerated fibers (P<0.05) and an increase in the number of preserved fibers (P<0.05) 4 days after injury. Additionally, most preserved fibers showed a normal G-ratio. These results indicated that calpain inhibition prevented the degeneration of optic nerve fibers, rescuing axons from the process of axonal degeneration. However, analysis of retinal ganglion cell survival demonstrated no difference between the calpain inhibitor- and vehicle-treated groups, suggesting that although the calpain inhibitor prevented axonal degeneration, it had no effect on RGC survival after optic nerve damage.

Neural Ablation and Regeneration in Pain Practice

A nerve block is an effective tool for diagnostic and therapeutic methods. If a diagnostic nerve block is successful for pain relief and the subsequent therapeutic nerve block is effective for only a limited duration, the next step that should be considered is a nerve ablation or modulation. The nerve ablation causes iatrogenic neural degeneration aiming only for sensory or sympathetic denervation without motor deficits. Nerve ablation produces the interruption of axonal continuity, degeneration of nerve fibers distal to the lesion (Wallerian degeneration), and the eventual death of axotomized neurons. The nerve ablation methods currently available for resection/removal of innervation are performed by either chemical or thermal ablation. Meanwhile, the nerve modulation method for interruption of innervation is performed using an electromagnetic field of pulsed radiofrequency. According to Sunderland's classification, it is first and foremost suggested that current neural ablations produce third degree peripheral nerve injury (PNI) to the myelin, axon, and endoneurium without any disruption of the fascicular arrangement, perineurium, and epineurium. The merit of Sunderland's third degree PNI is to produce a reversible injury. However, its shortcoming is the recurrence of pain and the necessity of repeated ablative procedures. The molecular mechanisms related to axonal regeneration after injury include cross-talk between axons and glial cells, neurotrophic factors, extracellular matrix molecules, and their receptors. It is essential to establish a safe, long-standing denervation method without any complications in future practices based on the mechanisms of nerve degeneration as well as following regeneration.

Transient lysosomal activation is essential for p75 nerve growth factor receptor expression in myelinated Schwann cells during Wallerian degeneration / 대한해부학회지

Myelinated Schwann cells in the peripheral nervous system express the p75 nerve growth factor receptor (p75NGFR) as a consequence of Schwann cell dedifferentiation during Wallerian degeneration. p75NGFR has been implicated in the remyelination of regenerating nerves. Although many studies have shown various mechanisms underlying Schwann cell dedifferentiation, the molecular mechanism contributing to the re-expression of p75NGFR in differentiated Schwann cells is largely unknown. In the present study, we found that lysosomes were transiently activated in Schwann cells after nerve injury and that the inhibition of lysosomal activation by chloroquine or lysosomal acidification inhibitors prevented p75NGFR expression at the mRNA transcriptional level in an ex vivo Wallerian degeneration model. Lysosomal acidification inhibitors suppressed demyelination, but not axonal degeneration, thereby suggesting that demyelination mediated by lysosomes may be an important signal for inducing p75NGFR expression. Tumor necrosis factor-alpha (TNF-alpha) has been suggested to be involved in regulating p75NGFR expression in Schwann cells. In this study, we found that removing TNF-alpha in vivo did not significantly suppress the induction of both lysosomes and p75NGFR. Thus, these findings suggest that lysosomal activation is tightly correlated with the induction of p75NGFR in demyelinating Schwann cells during Wallerian degeneration.

The Role of Axonopathy in Parkinson's Disease

New genetic and environmental studies of Parkinson's disease have revealed early problems in synaptic function and connectivity indicating that axonal impairment may be an important hallmark in this disorder. Since many studies suggest that axonal dysfunction precedes cell body loss, it is critical to target axons with treatments aimed at preserving "connectivity" as well as to develop and verify "biomarkers" with which to assess disease progression and drug efficacy.

Wallerian Degeneration of the Brain and Spinal Cord After Traumatic Brain Injury

No abstract available.

Estudo da regeneração simpática pós simpaticotomia seletiva experimental (ramocotomia) / Study of sympathetic regeneration post experimental selective sympathicotomy (ramicotomy)

Introdução: A simpatectomia torácica é o único tratamento, definitivo e eficaz, para a hiperidrose primária. A ramicotomia é um procedimento cirúrgico tão eficaz, mais conservador e com menos efeitos adversos que a simpatectomia convencional, contudo foi abandonada pela alta taxa de recidiva, atribuída, até então, à secção incompleta dos ramos comunicantes, ao desenvolvimento de outras vias de condução para o estímulo central e à regeneração neural. A avaliação histológica dos ramos comunicantes simpáticos após a ramicotomia, pode ajudar a entender o processo de recidiva dos sintomas da hiperidrose e, dessa forma contribuir para o desenvolvimento de estratégias para evitá-la. MATERIAL E MÉTODOS: 28 suínos foram submetidos à ramicotomia por videotoracospia e divididos randomicamente em 5 grupos, sacrificados com 15, 45, 90, 135 e 180 dias de pós-operatório (DPO). Os segmentos operados foram removidos cirurgicamente e submetidos à avaliação macroscópica da regeneração assim como análise histológica dos ramos comunicantes brancos e cinzentos para quantificação da reação inflamatória, deposição de fibras de colágeno grossas e finas, fibras reticulares e células de Schwann por imuno-histoquímica. Os dados foram comparados ao grupo controle, composto por segmentos intactos, não operados. RESULTADOS: Não houve regeneração macroscópica no grupo de 15 DPO sendo presente em 41,6% dos casos no grupo 180 DPO (p < 0,05). A reação inflamatória foi determinante no processo de degeneração Walleriana, com presença importante das células de Schwann nos ramos pré-ganglionares (p < 0,05), as células de Schwann apresentaram evolução semelhante nos dois ramos a partir do grupo de 45DPO, mantendo-se em menor número nos ramos cinzentos. As fibras de colágeno foram cruciais na cicatrização e as fibras reticulares importantes na regeneração neural, com correlação negativa entre elas (r = - 0,414; p < 0,01)...

INTRODUCTION: Thoracic sympathectomy is the only definitive and efficient treatment for primary hyperhidrosis. The ramicotomy is a surgical procedure that is as efficient as conventional sympathectomy but more conservative, having less adverse effects then conventional sympathectomy. This procedure was abandoned on account of the high recurrence rate, attributed to the incomplete section of the rami communicantes and to the development of new pathways of conduction to the central stimuli. MATHERIAL AND METHODS: Twenty-eight swine underwent bilateral videothoracoscopic ramicotomy and were randomly divided into 5 groups. The animals were sacrificed at 15, 45, 90, 135 and 180 days post-operative POD. The segments were removed and evaluated for macroscopic regeneration and histological analysis of the white and gray rami communicantes analyzing the inflammatory reaction, deposition of thin and thick collagen fibers, reticular fibers and Schwann cells. The data was compared to intact segments of sympathetic trunk as a positive control. RESULTS: There was neither macroscopic nor microscopic regeneration at the 15 POD group. The remaining groups had an average of 41,6% of regeneration, more significant at the 180 POD group (p<0.05). The inflammatory reaction was crucial in the process of Wallerian degeneration, with an important participation of the Schwann cells in the pre-ganglionic rami (p<0.05). The Schwann cells presented a similar evolution in both rami beginning at the 45 POD group, with a smaller count in the gray rami. The collagen fibers were significant in the cicatrization and the reticular fibers were important in neural regeneration, with a meaningful negative correlation between them (r = - 0,414; p < 0,01)...

The Role of the Peripheral Chemokine, CCL3, in Hyperalgesia following Peripheral Nerve Injury in the Rat / 대한통증학회지

BACKGROUND: Upregulation of one type of the pro-inflammatory chemokine (CCL2) and its receptor (CCR2) following peripheral nerve injury contributes to the induction of neuropathic pain. Here, we examined whether another type of chemokine (CCL3) is involved in neuropathic pain. METHODS: We measured changes in mechanical and thermal sensitivity in the hind paws of naive rats or rats with an L5 spinal nerve ligation (SNL) after intra-plantar injection of CCL3 or met-RANTES, an antagonist of the CCL3 receptor, CCR1. We also measured CCL3 levels in the sciatic nerve and the hind paw skin as well as CCR1 expression in dorsal root ganglion (DRG) cells from the lumbar spinal segments. RESULTS: Intra-plantar injection of CCL3 into the hind paw of naive rats mimicked L5 SNL-produced hyperalgesia. Intra-plantar injection of met-RANTES into the hind paw of rats with L5 SNL attenuated hyperalgesia. L5 SNL increased CCL3 levels in the sciatic nerve and the hind paw skin on the affected side. The number of CCR1-positive DRG cells in the lumbar segments was not changed following L5 SNL. CONCLUSIONS: Partial peripheral nerve injury increases local CCL3 levels along the degenerating axons during Wallerian degeneration. This CCL3 binds to its receptor, CCR1, located on adjacent uninjured afferents, presumably nociceptors, to induce hyperalgesia in the neuropathic pain state.

Copper (Cu2+) induces degeneration of dopaminergic neurons in the nigrostriatal system of rats / 神经科学通报·英文版

<p><b>OBJECTIVE</b>To study the effects of intranigral injection of different doses of CuSO4.5H2O on dopaminergic neuron in the nigrostriatal system of rats.</p><p><b>METHODS</b>Wistar rats were divided into four groups, including control group, 10 nmol, 50 nmol and 200 nmol copper injected into left substantia nigra (SN) groups. Seven days after the intranigral injection of copper, dopamine (DA) contents in the striatum (Str) were measured by high performance lipid chromotophotography (HPLC); the density of tyrosine hydroxylase (TH) positive axons in the Str was measured by TH staining method; TH and Caspase-3 mRNA expression in the SN were measured by semi-quantitative RT-PCR. We detected the activity of superoxide dismutase (SOD) in the lesioned midbrain of rats using biochemical methods.</p><p><b>RESULTS</b>DA and its metabolites contents had no significant difference between control group and low dose (10 nmol) copper group. But from 50 nmol copper group, DA contents in the lesioned sides were reduced with the increase in the copper doses injected, showing a significant linear correlation (F = 34.16, P < 0.01). In the 50 nmol copper group, TH positive axons in the Str decreased compared with those of the control and unlesioned sides (F = 121.9, P < 0.01). In the 50 nmol copper group, TH mRNA expression decreased (t = 3.12, P < 0.01) while Caspase-3 mRNA expression increased (t = 8.96, P < 0.01) in the SN compared with the control. SOD activity decreased in the midbrain of rats treated with 50 nmol copper compared with that of the control (t = 2.33, P < 0.01).</p><p><b>CONCLUSION</b>Copper could induce damage of dopaminergic neurons in the SN of rats through destroying antioxidant defenses and promoting apoptosis.</p>

The Effects of Soybean Diet on the Diabetic Neuropathy in the Rats / 대한해부학회지

Neuropathy is a serious and disabling complication that contributes to increased morbidity and mortality in diabetic patients. There is progressive distal to proximal axonal atrophy that ultimately leads to Wallerian degeneration. This study was performed to identify the effect of soy bean on diabetic neuropathy using morphometry. Male Sprague-Dawley rats were grouped into control, diabetic with red chow diet and diabetic with soy bean diet. The myelinated nerve fibers were counted and fiber size distributions were evaluated in each group at 4 and 8 weeks, respectively. Diabetic neuropathy didn't develop in streptozotocin-induced diabetic rats at four weeks. At 8 weeks, the myelinated nerve fiber in diabetic with soy bean diet was larger in number than that in diabetic to which did not be administered insulin. The number of myelinated nerve fiber was not different between diabetic group with insulin and without insulin. Mean myelinated nerve fiber size was smaller in diabetic with soy bean diet than diabetic with red chow diet and control. Histogram of fiber size distribution was shifted to left in diabetic with red chow and soy bean diet groups compared to control. Light and electron microscopic findings showed marked degeneration of nerve fibers in diabetic with red chow diet but preservation in diabetic with soy bean diet. The level of glucose and HbA1c was lower in diabetic with soy bean diet than red chow diet. Soy bean could be effective in the protection of neuropathy induced by diabetes mellitus.

Ultrastructural changes of compressed lumbar ventral nerve roots following decompression

To study whether there will be a permanent lumbar nerve root scarring or degeneration secondary to continuous compression followed by decompression on the nerve roots, which can account for postlaminectomy leg weakness or back pain. The study was performed at the Department of Anatomy, Faculty of Medicine, King Abdul-Aziz University, Jeddah, Kingdom of Saudi Arabia during 2003-2005. Twenty-six adult male New Zealand rabbits were used in the present study. The ventral roots of the left fourth lumbar nerve were clamped for 2 weeks then decompression was allowed by removal of the clips. The left ventral roots of the fourth lumbar nerve were excised for electron microscopic study. One week after nerve root decompression, the ventral root peripheral to the site of compression showed signs of Wallerian degeneration together with signs of regeneration. Schwann cells and myelinated nerve fibers showed severe degenerative changes. Two weeks after decompression, the endoneurium of the ventral root showed extensive edema with an increase in the regenerating myelinated and unmyelinated nerve fibers, and fibroblasts proliferation. Three weeks after decompression, the endoneurium showed an increase in the regenerating myelinated and unmyelinated nerve fibers with diminution of the endoneurial edema, and number of macrophages and an increase in collagen fibrils. Five and 6 weeks after decompression, the endoneurium showed marked diminution of the edema, macrophages, mast cells and fibroblasts. The endoneurium was filled of myelinated and unmyelinated nerve fibers and collagen fibrils. Decompression of the compressed roots of a spinal nerve is followed by regeneration of the nerve fibers and nerve recovery without endoneurial scarring

The Usefulness of Diffusion Tensor MRI for the Prediction of Clinical Outcome in Patients with Acute Subcortical Infarction

BACKGROUND: Diffusion tensor MRI (DTI) is a new imaging technique and enables us to analyze the structural damage of fiber pathways and to monitor the time course of Wallerian degeneration of the pyramidal tract in stroke patients. We used DTI to investigate structural changes of the infarct area and the associated descending corticospinal tract in patients with subcortical infarct. METHODS: We examined 24 consecutive patients who presented with acute single cerebral infarct in the subcortical area and who also had undergone an MRI study within 7 days after symptom onset. Clinical outcome was assessed using the National Institutes of Health Stroke Scale (NIHSS) at admission, 7 days, 14 days and 30 days and modified Rankin Scale (mRS) at admission and 30 days. Each of the indices was achieved by post processing the acquired DTI data and correlated with the NIHSS. RESULTS: In infarct region, fractional anisotropy (FA) was significantly decreased compared with matched-contralateral regions (0.39 vs. 0.53, p<0.001). In the distal to the infarct, FA was significantly decreased at internal capsule (0.62 vs. 0.64, p=0.019), not at pons (0.51 vs. 0.53, p=0.103). The decrease of anisotropy at infarct region correlated positively with the NIHSS at 7, 14 and 30 days and mRS at 30 days after stroke, but the decrease of anisotropy at internal capsule did not correlate with the NIHSS. CONCLUSIONS: This study shows the potential of DTI to detect and monitor the structural degeneration of fiber pathways and to establish the prognosis in patients with acute subcortical cerebral infarct.

Degeneração Walleriana de origem vascular em ressonância magnética de paciente com paralisia supranuclear progressiva provável: fator etiológico ou associação fortuita? / Vascular Wallerian degeneration on magnetic resonance in a patient with probable progressive supranuclear palsy: aetiology or casual link?

Progressive supranuclear palsy (PSP) is one of the most important causes of parkinsonism non responsive to therapy. Vascular parkinsonism is not uncommon. However, the cause-effect relationship between them is uncertain. We report on a 65 year old man with probable PSP who developed the clinical features of the disease after a ischaemic stroke. Magnetic resonance imaging disclosed a corticospinal tract Wallerian degeneration. There is not such an observation in the literature about this possible correlation.

Paralisia supranuclear progressiva (PSP) é uma das principais causas de parkinsonismo-plus não responsivo a terapia. A ocorrência de doença cerebrovascular associada a parkinsonismo não é infreqüente, no entanto é difícil estabelecer a relação causa-efeito entre ambas. Relatamos o caso de um paciente de 65 anos com PSP provável iniciada após infarto cerebral, em que a imagem por ressonância magnética evidenciou sinais de degeneração walleriana do trato córtico-espinhal. Não há relato na literatura pesquisada sobre esta possível correlação.

Double-stranded RNA Induces Inflammatory Gene Expression in Schwann Cells: Implication in the Wallerian Degeneration

Schwann cells play an important role in peripheral nerve regeneration. Upon neuronal injury, activated Schwann cells clean up the myelin debris by phagocytosis, and promote neuronal survival and axon outgrowth by secreting various neurotrophic factors. However, it is unclear how the nerve injury induces Schwann cell activation. Recently, it was reported that certain cytoplasmic molecules, which are secreted by cells undergoing necrotic cell death, induce immune cell activation via the toll-like receptors (TLRs). This suggests that the TLRs expressed on Schwann cells may recognize nerve damage by binding to the endogenous ligands secreted by the damaged nerve, thereby inducing Schwann cell activation. Accordingly, this study was undertaken to examine the expression and the function of the TLRs on primary Schwann cells and iSC, a rat Schwann cell line. The transcripts of TLR2, 3, 4, and 9 were detected on the primary Schwann cells as well as on iSC. The stimulation of iSC with poly (I: C), a synthetic ligand for the TLR3, induced the expression of TNF-alpha and RANTES. In addition, poly (I: C) stimulation induced the iNOS expression and nitric oxide secretion in iSC. These results suggest that the TLRs may be involved in the inflammatory activation of Schwann cells, which is observed during Wallerian degeneration after a peripheral nerve injury.

Schwann cell apoptosis in Wallerian-degenerated sciatic nerve of the rat / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To investigate systematically Schwann cell apoptosis in Wallerian-degenerated sciatic nerve of the rat, and evaluate its time-related feature.</p><p><b>METHODS</b>Ninety-five SD rats were divided randomly into one normal group (8 rats) and 11 experimental groups (66 rats, 6 in each). Both hind legs of each rat in experimental groups were randomly divided into test leg (sciatic nerve transected) and control one (nerve uninjured). All test legs constituted a test group and all control legs constituted a control one. After operation, all rats were respectively sacrificed at 1 h, 6 h, 12 h, 24 h, 2 d, 3 d, 4 d, 8 d, 14 d, 21 d, and 30 d. We analyzed the specimens of mid-distal sciatic nerve, especially the morphological changes of the nerve, the different expression levels of S-100 protein and apoptosis-related proteins such as Bcl-2, Bax, and Fas in Schwann cells. The TUNEL method was used to detect the apoptotic rate of Schwann cells.</p><p><b>RESULTS</b>(1) The test group showed Wallerian degeneration. The number of Schwann cells began to decrease at 24 h, obviously decreased on day 3 and 4, then began to increase from day 8 and formed Bungner belt after 14 days. (2) Schwann cells generally expressed S-100 at a low level in all groups. The control group was not significantly different from the normal group. The test group had statistical significance at 1 h and day 21. (3) As an inhibitory gene protein of Schwann cell apoptosis, Bcl-2 positive rates in the control and test groups apparently elevated and were statistically different from the normal group. (4) As a promotive gene protein of Schwann cell apoptosis, the control and test groups expressed Bax at a high level and were statistically different from the normal group. (5) As a promotive gene protein of Schwann cell apoptosis, Fas positive rate in control group was slightly elevated, but had no statistical significance compared with the normal group. Fas positive rate in test group continuously elevated in a fluctuant way, with highly statistical significance compared with the normal group. (6) TUNEL detection further proved that Schwann cell apoptosis rarely existed in the normal group, and the left sciatic nerve had no statistical significance compared with the right sciatic nerve. While the test group showed lots of apoptotic nuclei at 6 h, 2 d, 4 d, and 21 d. It had highly statistical significance compared with the normal group.</p><p><b>CONCLUSIONS</b>Schwann cell apoptosis does exist in Wallerian-degenerated sciatic nerve of the rat after transection. Schwann cell apoptosis and its apoptotic genes expression have a time-related feature.</p>

Watery and dark axons in Wallerian degeneration of the opossum's optic nerve: different patterns of cytoskeletal breakdown?

In this paper we report a qualitative morphological analysis of Wallerian degeneration in a marsupial. Right optic nerves of opossums Didelphis marsupialis were crushed with a fine forceps and after 24, 48, 72, 96 and 168 hours the animals were anaesthetized and perfused with fixative. The optic nerves were immersed in fixative and processed for routine transmission electron microscopy. Among the early alterations typical of axonal degeneration, we observed nerve fibers with focal degeneration of the axoplasmic cytoskeleton, watery degeneration and dark degeneration, the latter being prevalent at 168 hours after crush. Our results point to a gradual disintegration of the axoplasmic cytoskeleton, opposed to the previous view of an "all-or-nothing'' process (Griffin et al 1995). We also report that, due to an unknown mechanism, fibers show either a dark or watery pattern of axonal degeneration, as observed in axon profiles. We also observed fibers undergoing early myelin breakdown in the absence of axonal alterations