雏鸡核黄素缺乏症的外周神经酶细胞化学

为进一步探讨核黄素缺乏时外周神经变化的发生机理,对琥珀酸脱氢酶、酸性磷酸酶、腺苷三磷酸酶、碱性磷酸酶进行了酶细胞化学研究。结果表明,核黄素不足的鸡,雪旺氏细胞和轴突中琥珀酸脱氢酶活性下降,酸性磷酸酶活性明显升高,定位于雪旺氏细胞膜上的腺苷三磷酸酶和碱性磷酸酶活性均下降。通过对琥珀酸脱氢酶、腺苷三磷酸酶、碱性磷酸酶活性变化的动态观察,证明轻症鸡自愈的现象以及雪旺氏细胞中琥珀酸脱氢酶活性下降可能是形成脱髓鞘、外周神经水肿、导致雪旺氏细胞膜上的腺苷三磷酸酶和碱性磷酸酶活性下降的原因之一,而增生的雪旺氏细胞中琥珀酸脱氢酶活性增强则可能是修复损伤的一种代偿。腺苷三磷酸酶活性下降与神经水肿密切相关。     

Increased phosphorylation of c-Jun NH (2)-terminal protein kinase in the sciatic nerves of Lewis rats with experimental autoimmune neuritis

The phosphorylation of c-Jun NH (2)-terminal protein kinase (JNK), one of the mitogen-activated protein kinases, was analyzed in the sciatic nerves of Lewis rats with experimental autoimmune neuritis (EAN). Western blot analysis showed that the expression levels of both phosphorylated JNK1 (p-JNK1, approximately 46 kDa) and phosphorylated JNK2 (p-JNK2, approximately 54 kDa) in the sciatic nerves of rats with EAN increased significantly (p < 0.05) at day 14 post-immunization (PI) and remained at this level at days 24 and 30 PI, with a slight decrease. In EAN-affected sciatic nerves, there was intense immunostaining for p-JNK in the infiltrating inflammatory cells (especially ED1-positive macrophages) and Schwann cells on days 14-24 PI, compared with those of controls. Some macrophages with increased p-JNK immunoreactivity was shown to be apoptotic, while some Schwann cells remained survived in this rat EAN model, suggesting that JNK is differentially involved in the EAN-affected sciatic nerves. These findings suggest that JNK phosphorylation is closely associated with the clearance of inflammatory cells as well as the activation of Schwann cells in the EAN affected sciatic nerves.     

Strain differences in the branching of the sciatic nerve in rats

The sciatic nerve in the rat is the site most often used for peripheral nerve regeneration studies. The length of sciatic nerve available for research, however, depends on the point at which the sciatic nerve divides into the peroneal and tibial nerves. In the present study, the hind limbs of 150 adult male rats of five different strains (Sprague-Dawley, Fischer 344, Wistar-Han, Lewis and Nude) were analysed with regard to femur length, the point at which the sciatic nerve divides into the tibial and peroneal nerves, and where these are surrounded by the same epineurium, and the point at which they are encased in individual epineurial sheaths. The results indicate that the lengths of sciatic nerve are fairly constant in all strains of rats. In absolute terms, they amount to about one-third of the length of the femur for stretches of undivided sciatic nerve, and up to nearly half of the femur length for stretches where the tibial and peroneal nerves are already present, but are still enclosed by the same epineurium. In 61.7% of the hind limbs examined in Fischer rats, however, no sciatic nerve could be seen as such, but only in the form of its successors surrounded by the separate epineuria. This makes it highly advisable not to use male adult Fischer rats in peripheral nerve regeneration studies with the sciatic nerve as the point of focus.     

雏鸡“卷趾”麻痹症坐骨神经病变观察

１７只１日龄肉用小公鸡,饲喂低水平核黄素日粮（１．６５ｍｇ／ｋｇ）,于２２－２３日龄间有２９．４％出现其型的“卷趾”麻痹症状。光镜观察,坐骨神经纤维雪旺氏细胞肿大、增生、酸性磷酶活性增强和髓鞘变性。严重病例的坐骨神经显著肿大,神经纤维分离,间质水肿和炎性细胞浸润。电镜观察,坐骨神经有髓神经纤维的髓鞘扭曲、分层、破碎、形成同心圆层状小体,神经轴膜与髓鞘分离形成间和神经轴索变形。雪旺氏细胞胞浆中见同心     

Tumor-specific diagnostic marker for transmissible facial tumors of Tasmanian devils: immunohistochemistry studies

Devil facial tumor disease (DFTD) is a transmissible neoplasm that is threatening the survival of the Tasmanian devil. Genetic analyses have indicated that the disease is a peripheral nerve sheath neoplasm of Schwann cell origin. DFTD cells express genes characteristic of myelinating Schwann cells, and periaxin, a Schwann cell protein, has been proposed as a marker for the disease. Diagnosis of DFTD is currently based on histopathology, cytogenetics, and clinical appearance of the disease in affected animals. As devils are susceptible to a variety of neoplastic processes, a specific diagnostic test is required to differentiate DFTD from cancers of similar morphological appearance. This study presents a thorough examination of the expression of a set of Schwann cell and other neural crest markers in DFTD tumors and normal devil tissues. Samples from 20 primary DFTD tumors and 10 DFTD metastases were evaluated by immunohistochemistry for the expression of periaxin, S100 protein, peripheral myelin protein 22, nerve growth factor receptor, nestin, neuron specific enolase, chromogranin A, and myelin basic protein. Of these, periaxin was confirmed as the most sensitive and specific marker, labeling the majority of DFTD cells in 100% of primary DFTD tumors and DFTD metastases. In normal tissues, periaxin showed specificity for Schwann cells in peripheral nerve bundles. This marker was then evaluated in cultured devil Schwann cells, DFTD cell lines, and xenografted DFTD tumors. Periaxin expression was maintained in all these models, validating its utility as a diagnostic marker for the disease.     

Age-related immunohistochemical and ultrastructural changes in rat oculomotor nerve

During ageing process, multiple changes occur on nervous tissue composed of cells and extracellular matrix. Changes on nervous tissues are usually known as degenerative changes on axon structure and connective tissue covering the nerve such as a decrease in the number of fibre or general structural changes. For this purpose, we have studied age-dependent ultrastructural changes in the rat oculomotor nerve with electron microscopy and also demonstrated collagen structure of the neural sheaths with immunohistochemical techniques. This study was conducted in Gazi University Faculty of Medicine, Department of Anatomy with a total of nine Wistar albino rats. We observed strong collagen type I immunoreactivity in endoneurium and slight to moderate reactivity in fibroblast cytoplasm in 3-month- and 12-month-old groups and mild reactivity in 24-month-old group. Collagen type IV immunoreactivity was stronger in endoneurium and perineurium in the 3-month- and 12-month-old groups compared with collagen type I and fibroblast cytoplasm showed a very strong reactivity. On the other hand, in the 24-month-old group, there was slight reactivity in endoneurium and a strong reactivity in perineurium. NGF staining showed moderate to strong reactivity on Schwann cells of the 3-month-old group. The immunoreactivity decreased in the 12-month- and 24-month-old groups. In the 3-month-old rat group, Schwann cell cytoplasm, mitochondrial structure and neurofilaments were normal. In the 12-month-old group, there were no changes in organelle distribution, mitochondrial structure and neurofilaments, but there was an increase in the connective tissue. An inconsiderable number of degenerated myelinated nerves were observed. We detected an important decrease in the collagen type I immunoreactivity, which could suggest that the endoneurium, perineurium and epineurium are less resistant to the age-related collagen loss and that the peripheral nerve is protected by a weaker barrier in the old group. The collagen type IV immunoreactivity was significantly decreased with age. NGF synthesis decreases with age because of Schwann cell structural degeneration or for different reasons. Thus, this could explain the diminished capacity of regeneration and damage of the myelination of the peripheral nerve.     

鸡马立克氏病外周神经的病理组织学研究

用光镜、电镜及特殊染色法对兰州某鸡场２０例自然发生的马立克氏病鸡进行了外周神经的病理组织学研究。结果表明,外周神经主要表现增生性和渗出性病变,前者以瘤细胞增生为主,伴以脱髓鞘和雪旺氏细胞增生;后者以浆液渗出为特征,并发生神经纤维轴索肿胀和髓鞘变化,瘤细胞浸润很少。本文首次报告了马立克氏病的内脏－皮肤－神经混合型病变,还探讨了瘤细胞增生与脱髓鞘、神经水肿之间的关系。     

Electrophysiologic studies of the cutaneous innervation of the pelvic limb of male dogs

The area of skin supplied by the afferent fibers in one cutaneous nerve is called the cutaneous area (CA) for that nerve. The CA of peripheral branches of lumbar and sacral spinal nerves responsive to the stimulation of hair follicle mechanoreceptors were mapped in 27 dogs. The amount of overlap among the CA was similar to that found for other CA of the body. The CA of peripheral branches of the sciatic nerve were restricted to the lateral, cranial, and caudal aspects of the pelvic limb distal to the stifle. The CA of the saphenous nerve was located on the medial side of the limb, except for a small area located on the lateral side of the crus. The distal part of the CA of the saphenous nerve was completely overlapped in the hind paw by branches of the superficial peroneal nerve laterally and the medial plantar branch of the tibial nerve medially. The CA for the deep peroneal nerve was located on the dorsal surface of the webbing between digits 2 and 3 and the adjacent skin of these digits. The CA of the plantar branches of the tibial nerve were small in comparison with the diameter of the nerve, suggesting that these branches contained nerve fibers supplying other, deeper structures in the hindpaw and that damage to these nerves would interfere with cutaneous sensation in only a small region on the plantar surface of the hindpaw. Knowledge of the CA of the various branches of the sciatic nerve allows more accurate localization of injury to the sciatic nerve or its branches by using areas of anesthesia.     

Electrophysiologic characteristics of tibial and sciatic nerves in the hen

The peripheral nerve of the hen has become an increasingly important animal model in studies of peripheral neuropathy, especially that induced by organophosphorus agent exposure. However, few electrophysiologic studies have been performed, and few data on normal peripheral nerve function exist. The purpose in the present study was to measure the characteristics of the compound action potential of peripheral nerves of the healthy hen. The results showed that conduction velocities of the tibial and the sciatic nerves were 41 m/s and 60 m/s, respectively. The relative refractory period was slightly shorter in the sciatic nerve. The tibial nerve exhibited a lower threshold for stimulation and a significantly shorter chronaxie (19 microseconds) than did the sciatic nerve (25 microseconds). Variability between animals was relatively small. These data should provide useful reference points on studies in which peripheral neuropathy is induced or suspected.     

Peripheral neuropathy associated with dietary riboflavin deficiency in the chicken. I. Light microscopic study

Chickens fed a riboflavin-deficient diet from hatching had leg weakness and paralysis as early as 12 days of age. Signs worsened through day 16; after 35 days, recovery was evident. Sciatic nerves from affected chickens were enlarged. Significant microscopic lesions were confined to peripheral nerves and included tissue separation (suggesting interstitial edema), Schwann cell swelling, perivascular leukocytic infiltration, and segmental demyelination accompanied by accumulation of osmiophilic debris in Schwann cell cytoplasm. Axon degeneration was present, but was not a primary lesion. Acid phosphatase enzyme activity of Schwann cells was increased in affected nerves. These results demonstrate that dietary riboflavin deficiency causes a demyelinating peripheral neuropathy in young, rapidly growing chickens.     

Acquired spinal cord and peripheral nerve disease.

Acquired spinal cord diseases in ruminants result most commonly from infectious, traumatic, metabolic/nutritional, or toxic causes and rarely from neoplasia. Clinical signs of spinal cord disease depend on the neuroanatomic location of the lesion. Acquired spinal cord diseases including vertebral osteomyelitis/spinal abscess, cauda-equina disease, enzootic ataxia, lymphosarcoma,polyradiculoneuritis, and degenerative myeloencephalopathy are discussed. Acquired peripheral nerve disease in cattle most often is a result of injury, and most commonly only one limb is involved.Peripheral nerve injuries frequently occur secondary to myopathy in recumbent adult cattle. In small ruminants, peripheral nerve injury seems less common, most likely due to their smaller size,but may occur from predator wounds or iatrogenically following intramuscular drug administration. Injury to the brachial plexus and radial, suprascapular, sciatic, femoral, and obturator nerves is discussed.     

Peripheral hypomyelinization in two golden retriever littermates

Peripheral hypomyelinization was found in 2 Golden Retriever littermates that had pelvic limb ataxia, depressed postural reactions, and depressed segmental reflexes. Diagnostic findings included infrequent denervation potentials, reduced or absent evoked potentials, and markedly diminished motor nerve conduction velocities. Light and electron microscopy of peripheral nerves revealed fewer than normal myelinated axons, myelinated sheaths inappropriately thin for the caliber of the fiber, poor myelin compaction, greater than normal numbers of Schwann cell nuclei, many Schwann cells with voluminous cytoplasm, and greater than normal amount of perineural collagen. Findings were compatible with a peripheral hypomyelinization process; a defect in Schwann cell function was suspected.     

Contrast-enhanced ultrasonography for evaluation of the blood perfusion of sciatic nerves in healthy dogs

Blood supply to the peripheral nerves is essential for fulfilling their structural and functional requirements. This prospective, experimental, exploratory study aimed to assess the feasibility of contrast-enhanced ultrasonography (CEUS) for evaluating blood perfusion of the sciatic nerve in normal dogs. Contrast-enhanced ultrasonography examinations were performed on the bilateral sciatic nerves after bolus injection of Sonazoid™ (0.015 mL/kg) in 12 healthy Beagles for 150 s. Then, qualitative assessment of the wash-in timing, degree and enhancement patterns, and quantitative measurement of the peak intensity and time to peak intensity were performed from the sciatic nerve. The results were compared to those obtained from the adductor muscle around the nerve and caudal gluteal artery. After contrast agent injection, the sciatic nerve was enhanced at approximately 13–14 s, immediately after wash-in of the caudal gluteal artery. The peak intensity of the sciatic nerve was significantly lower than that of the caudal gluteal artery and higher than that of the adductor muscle. The time to peak intensity was significantly slower than that of the caudal gluteal artery; but was not significantly different from that of the adductor muscle. There were no significant differences in the peak intensity and time to peak intensity between the left and right sciatic nerves. These results demonstrate the feasibility of CEUS to assess blood perfusion of the sciatic nerve in healthy dogs qualitatively and quantitatively. This result from healthy dogs could serve as a reference for further studies that evaluate the sciatic nerve under pathological conditions.     

Influence of electrical stimulation on regeneration of the radial nerve in dogs

The effects of biphasic electric fields on nerve regeneration that follows injury to the left radial nerve were studied in dogs by electromyography (EMG). Left and right radial nerves were crushed with a serrated haemostat. Stimulating electrodes were positioned proximally and distally to the site of the injury. The left nerves received rectangular, biphasic and current pulses (30 microA, 0.5 Hz) through the injury for two months. The right radial nerves were treated as controls and regenerated without electrical stimulation. EMG activities were recorded intramuscularly from the left and right musculus extensor digitalis communis (MEDC). Results obtained at the end of the two-month stimulation period showed a significant difference in EMG activity between the left (stimulated) and the right (non-stimulated) MEDC, suggesting that electrical treatment enhanced nerve regeneration.     

ULTRASONOGRAPHIC ASSESSMENT OF THE CANINE SCIATIC NERVE

To describe the ultrasonographic technique for investigation of the canine sciatic nerve, four canine cadaver pelvic limbs, two live healthy dogs, and five canine patients with suspected peripheral sciatic nerve lesions were examined with a high-resolution linear ultrasound transducer. The caudal part of the lumbosacral trunk and the origin of the sciatic nerve were visualized through the greater ischiatic foramen. The two components of the sciatic nerve, common peroneal and tibial nerves, were distinguished along the entire length of the nerve, until they branched at the level of the distal femur. In healthy live dogs they appeared as two adjacent hypoechoic tubular structures with internal echotexture of discontinuous hyperechoic bands, surrounded by a thin rim of highly echogenic tissue. The common peroneal component had a smaller diameter and was on the cranial aspect of the tibial component. An ultrasonographic lesion compatible with a peripheral nerve sheath tumor was found in one dog. Improved understanding of the ultrasonographic anatomy of the sciatic nerve supports clinical use of this modality.     

Anatomical and experimental studies of brachial plexus, sciatic, and femoral nerve-location using peripheral nerve stimulation in the dog

OBJECTIVE: To investigate the anatomy of the brachial plexus, sciatic, and femoral nerves for the use of a peripheral nerve-stimulator to perform nerve blocks in dogs. STUDY DESIGN: Prospective experimental trial. ANIMALS: Four canine cadavers and four healthy adult dogs weighing 23 +/- 2.5 kg. METHODS: Phase I: in four canine cadavers, an anatomical study was conducted to evaluate accurate needle-insertion techniques. Phase II: the utility of these techniques, and the value of electrostimulation, were evaluated in four anesthetized dogs in lateral recumbency (medetomidine, 5 microg kg(-1)/ketamine 5 mg kg(-1)) using an electrical stimulator and shielded needles. RESULTS: For the brachial plexus, the needle was inserted cranial to the acromion, medial to the subscapularis muscle, at an angle of approximately 20-30 degrees in relation to a plane vertical to the surface on which the animal was lying, oriented parallel to the long axis of the animal, in a ventro-caudal direction. For the sciatic nerve, the needle was inserted just cranial to the sacrotuberous ligament, through the gluteus superficialis muscle, at an angle of approximately 60 degrees in relation to the horizontal plane, in a ventro-cranial direction, and up to the level of the ischium. For the femoral nerve, the needle was inserted perpendicular to the skin, just cranial to the femoral artery, and directed a little caudally. Using a peripheral nerve-stimulator, all nerves were located, and muscle contractions were elicited at a current of 0.2-0.4 mA. No complications were observed during the procedures. CONCLUSION: Electrostimulation of peripheral nerves is useful in locating the branches of the brachial plexus as well as the sciatic and femoral nerves in dogs. CLINICAL RELEVANCE: Peripheral nerve stimulation increases the reliability of a nerve block when compared with blind needle-insertion.     

Propagation of scrapie in peripheral nerves after footpad infection in normal and neurotoxin exposed hamsters

As is known from various animal models, the spread of agents causing transmissible spongiform encephalopathies (TSE) after peripheral infection affects peripheral nerves before reaching the central nervous system (CNS) and leading to a fatal end of the disease. The lack of therapeutic approaches for TSE is partially due to the limited amount of information available on the involvement of host biological compartments and processes in the propagation of the infectious agent. The in vivo model presented here can provide information on the spread of the scrapie agent via the peripheral nerves of hamsters under normal and altered axonal conditions. Syrian hamsters were unilaterally footpad (f.p.) infected with scrapie. The results of the spatiotemporal ultrasensitive immunoblot-detection of scrapie-associated prion protein (PrP(Sc)) in serial nerve segments of both distal sciatic nerves could be interpreted as a centripetal and subsequent centrifugal neural spread of PrP(Sc) for this route of infection. In order to determine whether this propagation is dependent on main components in the axonal cytoskeleton (e.g. neurofilaments, also relevant for the component ;a' of slow axonal transport mechanisms), hamsters were treated -in an additional experiment- with the neurotoxin beta,beta-iminodiproprionitrile (IDPN) around the beginning of the scrapie infection. A comparison of the Western blot signals of PrP(Sc) in the ipsilateral and in the subsequently affected contralateral sciatic nerve segments with the results revealed from IDPN-untreated animals at preclinical and clinical stages of the TSE disease, indicated similar amounts of PrP(Sc). Furthermore, the mean survival time was unchanged in both groups. This in vivo model, therefore, suggests that the propagation of PrP(Sc) along peripheral nerves is not dependent on an intact neurofilament component of the axonal cytoskeleton. Additionally, the model indicates that the spread of PrP(Sc) is not mediated by the slow component ;a' of the axonal transport mechanism.     

Cutaneous saphenous nerve graft for the treatment of sciatic neurotmesis in a dog

CASE DESCRIPTION: A 2-year-old Griffon Vendéen was examined because of a 1-month history of right hind limb lameness after a traumatic injury. CLINICAL FINDINGS: Neurologic examination revealed monoplegia and anesthesia of the right hind limb distal to the stifle (femorotibial) joint except for the area supplied by the cutaneous saphenous nerve. Results of electromyographic testing were consistent with a severe lesion of the tibial and peroneal nerves at the level of the stifle joint. TREATMENT AND OUTCOME: Exploratory surgery revealed an 80-mm-long gap in both the peroneal and tibial branches of the right sciatic nerve. A section of the left cutaneous saphenous nerve was interposed to graft the nerve defects. The dog received joint mechanotherapy and electrophysiologic therapy during the reinnervation process. Ten months after surgery, the dog had recovered almost completely. Neurologic examination revealed diminished flexion of the tarsal and digital joints. Repeat electromyographic testing revealed no abnormal spontaneous electrical activity in the right hind limb musculature, and small compound muscle action potentials were recorded in the right interosseous and cranial tibial muscles. CLINICAL RELEVANCE: Without surgical treatment, neurotmesis injury results in poor recovery of motor and sensory functions and may result in amputation. If a nerve defect exists, nerve grafting should be considered, even if the procedure is delayed until well after the injury. The sensory portion of the cutaneous saphenous nerve is a potential source of peripheral nerve for grafting in dogs. Reinnervation is a long-term process and physiologic support and owner involvement are necessary, but nearly complete functional recovery is possible.     

Cytotoxicity and cytokine production by bovine alveolar macrophages challenged with wild type and leukotoxin-deficient Mannheimia haemolytica

The aim of this study was to investigate the use of ultrasound (US) guidance to perform sciatic and saphenous nerve blocks in dogs. Five dogs were sedated with medetomidine and butorphanol. A high-resolution US transducer was used to locate the nerves, guide placement of the needle and visualise the perineural injection of lidocaine 2%. Electrostimulation was used to confirm correct placement prior to the sciatic block. Nerve functions were evaluated over a 3 h period following administration of atipamezole. Successful identification of the nerves and the quality of the blocks were recorded. Location of the nerves, complete sensory block of the saphenous nerve, and partial to complete sensory and motor blocks of the sciatic nerve were achieved in all dogs. The resultant US guidance is potentially valuable for blocking the sciatic and saphenous nerves in dogs, although further work will be required to ensure a complete block of the sciatic nerve.     

嗅鞘细胞生物学特性及其在脊髓再生中应用的研究

近年来研究证明嗅球及嗅神经组织可以分离出嗅神经鞘细胞(olfactory ensheathing cells,OECs)，OECs具有中枢神经系统（CNS）星形胶质细胞和外周雪旺式细胞的特性，移植后的OCEs能够在宿主损伤的脊髓组织进行迁移，能够引导支持损伤的神经元突起再生、生长、延伸。而且还可以通过转基因技术使其携带某些促进神经元再生的外源性基因，表达一些促神经再生的分子，从而改善神经损伤的内环境，诱导损伤的脊髓神经元再生，使得受损脊髓再生的潜力得以发挥，达到脊髓再生和功能恢复的目的。本文就OECs的生物学特性及其在脊髓损伤和CNS的基因治疗进行综述。