GAP-43 expression in the developing rat lumbar spinal cord.

The expression of the growth-associated protein GAP-43, detected by immunocytochemistry, has been studied in the developing rat lumbar spinal cord over the period E11 (embryonic day 11), when GAP-43 first appears in the spinal cord, to P29 (postnatal day 29) by which time very little remains. Early GAP-43 expression in the fetal cord (E11-14) is restricted to dorsal root ganglia, motoneurons, dorsal and ventral roots and laterally positioned and contralateral projection neurons and axons. Most of the gray matter is free of stain. The intensity of GAP-43 staining increases markedly as axonal growth increases, allowing clear visualization of the developmental pathways taken by different groups of axons. Later in fetal life (E14-19), as these axons find their targets and new pathways begin to grow, the pattern of GAP-43 expression changes. During the period, GAP-43 staining in dorsal root ganglia, motoneurons, and dorsal and ventral roots decreases, whereas axons within the gray matter begin to express the protein and staining in white matter tracts increases. At E17-P2 there is intense GAP-43 labelling of dorsal horn neurons with axons projecting into the dorsolateral funiculus and GAP-43 is also expressed in axon collaterals growing into the gray matter from lateral and ventral white matter tracts. At E19-P2, GAP-43 is concentrated in axons of substantia gelatinosa. Overall levels decline in the postnatal period, except for late GAP-43 expression in the corticospinal tract, and by P29 only this tract remains stained.     

Redistribution of B-50/growth-associated protein 43 during differentiation and maturation of rat hippocampal neurons in vitro.

Morphologically polarized hippocampal neurons, grown in culture for two days, contain immunoreactivity of the growth-associated protein B-50 along the plasma membrane of both dendrites and axons. In mature hippocampal neurons, both in vitro and in vivo, B-50 is located in the axon. In order to assess at which stage during neuronal differentiation B-50 is selectively located in the axon, an immuno-light and electron-microscopic study was performed on rat hippocampal neurons developing in vitro. B-50 immunofluorescence was detected in the axon, dendrites and soma of two-day-old polarized neurons. Simultaneously, microtubule-associated protein 2, a marker specific to dendritic microtubules, was predominantly found in the soma, the short dendritic processes and at the base of axonal growth cones. In hippocampal neurons cultured beyond seven days in vitro, microtubule-associated protein 2 immunofluorescence is restricted to the cell soma and dendrites. The spatial distribution of B-50, however, varies. In solitary neurons maturing without interneuronal contacts, B-50 immunofluorescence is observed in axons and in the dendrosomatic domain characterized by the presence of microtubule-associated protein 2. In contrast, in high-density cell cultures B-50 immunofluorescence is absent in the cell body and dendrites, but punctate in axons running along the dendrites. Electron microscopy was carried out on hippocampal neurons of eight to 21 days in vitro to study the process of redistribution of B-50 at the subcellular level. In neurons of eight days in vitro with prominent synapses, B-50 immunoreactivity is significantly elevated at the axonal plasma membrane compared to the plasma membrane of the dendrites and the soma. In neurons from the same culture without synapses, B-50 immunoreactivity is distributed rather densely along the plasma membrane of the soma, dendrites, and on the axonal plasma membrane. A similar B-50 distribution is observed in mature neurons cultured at low cell density without interneuronal cell contacts, for 15 days in vitro. In high-density cell cultures of 21 days in vitro, B-50 is virtually absent at the plasma membrane of the soma and dendrites, and heterogenously distributed along the plasma membrane of axon and axonal varicosities. Our results indicate that selective sorting of B-50 into axons occurs after initial morphological polarization of hippocampal neurons and is correlated with the formation of synapses and with the cessation of dendritic outgrowth.     

The distribution of GAP-43 in normal rat spinal cord

Summary We have studied the distribution of the growth-associated protein GAP-43 in the spinal cord of adult rats by light and electron microscopy, using a new antiserum raised against GAP-43/-galactosidase fusion protein. We show that GAP-43 is present at all vertebral levels but is more concentrated in cervical and thoracic regions. In addition to heavy staining in the corticospinal tracts of the white matter, staining can be seen at the light microscopic level throughout the grey matter and is particularly heavy around the central canal and in the superficial dorsal horn. Electron microscopic examination revealed that GAP-43 immunostaining is confined to a subpopulation of axons and axon terminals. Staining occurs in small myelinated and unmyelinated fibres and in terminals which are mainly small and make single axodendritic or axosomatic synapses. Staining in such terminals occurs in the axoplasm but is heaviest immediately adjoining the axolema. Staining was not observed in dendrites, nor in large myelinated axons or large axon terminals. Our results indicate that GAP-43 is expressed in adult rat spinal cord in a subpopulation of small diameter fibres and axon terminals. The distribution and morphology of these terminals is consistent with several different possible origins including corticospinal projection neurons, small diameter primary afferent neurons, and descending raphe-spinal serotonin containing neurons.     

GAP-43 immunoreactivity is widespread in the autonomic neurons and sensory neurons of the rat.

GAP-43 is a membrane-bound phosphoprotein generally associated with axon growth during development and regeneration. Using immunohistochemical and immunoblotting techniques this study shows that GAP-43 is expressed extensively in the unperturbed adult autonomic nervous system. Strong immunoreactivity was seen in the developing and mature enteric subdivision of the autonomic nervous system and in nerves of the iris and various blood vessels. The presence of GAP-43 immunoreactivity in varicose nerve fibres, and a comparison of the labelling pattern of GAP-43 with the nerve associated marker PGP 9.5 suggests that GAP-43 is present in most or all autonomic nerve fibres in these organs. Immunoblotting of gut samples on 10% polyacrylamide gels revealed a single band of approximately 45,000 mol. wt that co-migrated with pure central nervous system GAP-43. Surgical sympathectomy experiments resulting in almost complete elimination of sympathetic fibres did not markedly affect the pattern of GAP-43 immunoreactivity in the iris, indicating that GAP-43 is expressed not only in sympathetic nerves but also in parasympathetic and sensory fibres. These findings show that GAP-43 is expressed extensively in autonomic nerves of the adult rat, at levels comparable to those seen during development. High levels of GAP-43 are not therefore restricted to development and regeneration in this part of the nervous system.     

Immunocytochemistry of B-50 (GAP-43) in the spinal cord and in dorsal root ganglia of the adult cat

Summary The distribution of the neural-specific growth associated protein B-50 (GAP-43), which persists in the mature spinal cord and dorsal root ganglia, has been studied by light and electron microscopic immunohistochemistry in the cat. Throughout the spinal cord, B-50 immunoreactivity was seen confined to the neuropil, whereas neuronal cell bodies were unreactive. The most conspicuous immunostaining was observed in the dorsal horn, where it gradually decreased from superficial laminae (I–II) toward more ventral laminae (III–V), and in the central portion of the intermediate gray (mainly lamina X). In these regions, the labelling was localized within unmyelinated, small diameter nerve fibres and axon terminals. In the rest of the intermediate zone (laminae VI–VIII), B-50 immunoreactivity was virtually absent. The intermediolateral nucleus in the thoracic and cranial lumbar cord showed a circumscribed intense B-50 immunoreactivity brought about by the labelling of many axon terminals on preganglionic sympathetic neurons. In motor nuclei of the ventral horn (lamina IX), low levels of B-50 immunoreactivity were present in a few axon terminals on dendritic and somal profiles of motoneurons. In dorsal root ganglia, B-50 immunoreactivity was mainly localized in the cell bodies of small and medium-sized sensory neurons. The selective distribution of persisting B-50 immunoreactivity in the mature cat throughout sensory, motor, and autonomie areas of the spinal cord and in dorsal root ganglia suggests that B-50-positive systems retain in adult life the capacity for structural and functional plasticity.     

大鼠羊膜上皮细胞移植对损伤脊髓生长相关蛋白43表达的影响

目的:探讨羊膜上皮细胞(AECs)移植对大鼠损伤脊髓生长相关蛋白43(GAP-43)表达和神经功能恢复的影响.方法: 取E12d～14d的SD大鼠AECs体外培养;改良-Allen撞击法制备大鼠脊髓损伤(SCI)模型,并随机分成假手术组、SCI+生理盐水对照组和SCI+AECs移植组;分别于术后第1、3、7、14和28天通过BBB 神经行为学评分法对神经功能进行评估,观察其恢复状况,并采用免疫组织化学和免疫印迹法检测脊髓组织GAP-43表达的变化.结果: GAP-43的表达于脊髓损伤后第7天显著增加,AECs移植后损伤脊髓中的GAP-43持续高表达至第28天,而盐水对照组的表达于术后14d左右逐渐恢复至正常水平.与盐水对照组相比较,AECs移植后2～4周脊髓损伤大鼠的后肢运动功能得到一定的改善.结论: AECs移植后可使损伤大鼠脊髓GAP-43的表达增高,并在一定程度上促进了大鼠后肢运动功能的恢复.     

Ultrastructural evidence for the lack of co-transport of B-50/GAP-43 and calmodulin in myelinated axons of the regenerating rat sciatic nerve

Summary Following peripheral nerve injury, neurons respond with synthesis of proteins required for axonal regeneration. Newly synthesized membrane proteins, like B-50/GAP-43, are transported with the fast component of anterograde axonal transport. Structural proteins and calmodulin are transported by the slow component. Since B-50/GAP-43 can bind calmodulin, it has been hypothesised that B-50/GAP-43 may act as a carrier for fast anterograde transport of calmodulin, so that both proteins are delivered rapidly to the distally outgrowing axons (the fast carrier hypothesis). We have investigated whether this hypothesis is valid in myelinated axons of the regenerating rat sciatic nerve. Seven days after crush, the nerve was ligated to accumulate fast transported proteins. Nerve pieces were dissected proximal to the ligation and processed for immunofluorescence and quantitative electron microscopy by postembedding single and double immunogold labelling. By light microscopy, we observed a qualitative increase in B-50/GAP-43 immunofluorescence in the axonal element immediately proximal to the nerve ligation (termed accumulated) compared to an upstream site (termed regenerating) closer to the cell body. The immunofluorescence for calmodulin appeared to be the same at both sites. Using electron microscopy, we observed that organelles had collected at the accumulated site, moreover the density of B-50/GAP-43 immunolabelling was significantly increased compared to the regenerating site, where the axoplasmic structure was undisturbed. The increase in B-50/GAP-43 immunolabelling was largely associated with vesicles. The density of calmodulin immunolabelling was similar at both sites. Approximately 25% of the total B-50/GAP-43 was associated with vesicles of which only 15% also contained labelling for calmodulin. Thus, ligation of the nerve resulted in accumulation of vesicles, including those carrying B-50/GAP-43, largely without calmodulin. Therefore, contrary to the fast carrier hypothesis, the bulk of calmodulin is not co-transported with B-50/GAP-43 in myelinated axons of the sciatic nerve.     

Mapping the development of the rat brain by GAP-43 immunocytochemistry.

Growth-associated protein-43 (GAP-43) is a phosphoprotein of the nerve terminal membrane which has been linked to the development and restructuring of axonal connections. Using a monospecific antibody prepared in sheep against purified GAP-43, we examined the temporal and spatial changes in the distribution of this protein from embryonic stage day 13 (E13) to adulthood. At stages in which neurons are still dividing and migrating, levels of GAP-43 are extremely low, as is seen in the cortical plate throughout the embryonic period. With the onset of process outgrowth, intense GAP-43 immunoreactivity appears along the length of axons: by E13, such staining is already strong in the brainstem, where it continues up through the first postnatal week and then disappears. In the neocortex, intense fiber staining first appears several days later but ends at the same time as in the brainstem. At the end of the period of intense axonal staining there is a brief interval in which high levels of GAP-43 immunostaining are seen in the neuropil. In regions of the brain in which specific developmental events have been characterized anatomically and physiologically, the period of dense neuropil staining coincides with the formation of axonal end-arbors, the beginning of synaptogenesis, and the time at which synaptic organization can be modified by the impingent pattern of activity (i.e. the critical period). Over the next few days, staining in neuropil declines sharply in most regions except for certain structures in the rostral neuraxis which may be sites of ongoing synaptic remodeling.     

Distribution of growth-associated protein, B-50 (GAP-43) in the mammalian enteric nervous system

The presence of the growth-associated protein, B-50 (also known as GAP-43) was investigated in the adult mammalian enteric nervous system. The small intestine of rat, ferret and human was examined by immunohistochemistry. Dense B-50-like immunoreactivity was localized in nerves throughout the wall of the rat, ferret and human small intestine, notably in the myenteric and submucous plexuses, where in the ferret ileum it co-localized with vasoactive intestinal polypeptide-immunoreactive fibre groups. Material with the biochemical and immunological characteristics of rat B-50 was extracted from the rat ileum. In-situ hybridization demonstrated that enteric neurons express B-50. These findings are consistent with a role for B-50 in the documented plasticity of the adult enteric nervous system.     

Expression of GAP-43 during development and after monocular enucleation in the rat superior colliculus

The retinotectal projection of rodents presents a precise retinotopic organization that develops, from diffuse connections, from the day of birth to post-natal day 10. Previous data had demonstrated that these projections undergo reorganization after retinal lesions, nerve crush and monocular enucleation. The axonal growth seems to be directly related to growth-associated protein-43 (GAP-43) expression, a protein predominantly located in growth cones, which is regulated throughout development. GAP-43 is presented both under non-phosphorylated and phosphorylated (pGAP-43) forms. The phosphorylated form, has been associated to axon growth via polymerization of F-actin, and synaptic enhancement through neurotransmitter release facilitation. Herein we investigated the spatio-temporal expression of GAP-43 in the rat superior colliculus during normal development and after monocular enucleation in different stages of development. Lister Hooded rats ranging from post-natal day 0 to 70 were used for ontogeny studies. Another group of animals were submitted to monocular enucleation at post-natal day 10 (PND10) or PND21. After different survival-times, the animals were sacrificed and the brains processed for either immunohistochemistry or western blotting analysis. Our data show that GAP-43 is expressed in retinotectal axons in early stages of development but remains present in adulthood. Moreover, monocular enucleation leads to an increase in pGAP-43 expression in the deafferented colliculus. Taken together these results suggest a role for pGAP-43 in retinotectal morphological plasticity observed both during normal development and after monocular enucleation.     

ATP诱导大鼠原代培养海马神经元神经毒性及对GAP-43表达的影响

目的:观察不同浓度ATP对大鼠原代培养海马神经元的损伤作用及GAP-43表达的影响,并探讨其可能机制。方法:原代培养新生SD大鼠海马神经元至第8 d,MTT法检测不同浓度ATP作用下海马神经元的存活率,倒置相差显微镜观察海马神经元的形态学改变,免疫荧光细胞化学法观察损伤后海马神经元GAP-43的表达变化,钙离子荧光染料Flou-4/AM检测细胞内钙离子的浓度变化。结果:低浓度ATP对海马神经元无明显影响,高浓度ATP对海马神经元有毒性损伤作用,且呈剂量依赖性。P2X受体拮抗剂可阻断ATP介导的细胞毒性作用,但P2X7受体拮抗剂的阻断作用不明显。高浓度ATP作用海马神经元4 h后,GAP-43的表达水平明显上调,且表达部位发生改变,主要表达于胞体,突起表达减少甚至消失。高浓度ATP组荧光衰减速率明显减慢,说明高浓度ATP参与了海马神经元细胞内钙调节。结论:高浓度ATP对海马神经元有毒性损伤作用,损伤后GAP-43代偿性表达增多,且与细胞内钙离子浓度变化有关,P2X7受体可能不是介导ATP此作用的主要受体亚型。     

Immunolocalization of B-50 (GAP-43) in the mouse olfactory bulb: Predominant presence in preterminal axons

Summary Because the growth-associated protein B-50 (GAP-43) has been implicated in neurite outgrowth as well as in synaptic plasticity, we studied its light and electron microscopical distribution in the mouse olfactory bulb, an area of the nervous system which exhibits a high degree of synaptic plasticity. Immunofluorescent staining with monospecific affinity-purified anti-B-50 antibodies revealed that B-50 is most abundantly expressed in the olfactory nerve fibre layer and the granule cell layer neuropil, while little staining was observed in the external plexiform layer and in cell bodies. B-50 is absent from dendrites and myelinated axons as indicated by double labelling with monoclonal antibodies against microtubule-associated protein 2 and the large neurofilament protein, respectively. Using post-embedding immunogold labelling on ultrathin Lowicryl sections, B-50 was found to be highly concentrated in presumed growth cones in the olfactory nerve fibre layer and in thin unmyelinated axons and presynaptic terminals in the granule cell layer neuropil. Near background immunolabelling was seen in perikarya, dendrites and myelinated axons. In view of the implication of B-50 in plasticity-related phenomena, its abundance in the thin unmyelinated preterminal axons suggests that these are potential sites of extrasynaptic plasticity.     

Regionalized cadherin-7 expression by radial glia is regulated by Shh and Pax7 during chicken spinal cord development

During development, several genes that specify neuronal subtype identity are expressed in distinct dorsoventral domains of the spinal cord and hindbrain. Cadherin-7 (Cad7), a member of the cadherin family of adhesion molecules, is expressed by radial glia in a dorsal domain of the spinal cord basal plate in chicken. To study the regulation of the Cad7 gene, we ectopically expressed two known dorsoventral patterning genes, Shh and Pax7, in the caudal neural tube and in two brain regions at different stages of development by in vivo electroporation. Results showed that Shh regulated the expression of Cad7 by radial glia in a concentration-dependent manner. Shh induced or repressed the expression of Cad7, at low and high concentrations, respectively. Furthermore, Pax7 inhibited the expression of Cad7. These results are compatible with a role of Shh and Pax7 in regulating endogenous Cad7 expression during spinal cord and hindbrain development. Our data show, for the first time, that Shh can regulate the expression not only of other gene regulatory factors, but also of Cad7, a morphoregulatory molecule that plays a role in axon elongation and neural circuit formation.     

B-50/GAP43 localization in polarized hippocampal neurons in vitro: an ultrastructural quantitative study.

Hippocampal pyramidal neurons cultured in vitro gradually develop morphologically and biochemically distinct axons and dendrites, resulting in functional neuronal polarization [Dotti C. G. et al. (1988) J. Neurosci. 8, 1454-1468]. We have studied the distribution of the growth-associated protein B-50 in hippocampal neurons of the rat at stage 3 of development by means of light and electron microscopic immunocytochemistry. Hippocampal neurons grown for two to three days in vitro were aldehyde fixed and immunolabelled using polyclonal rabbit antibodies to B-50 and goat anti-rabbit immunoglobulins tagged with 1 nm gold particles. In order to permit visualization by both light and electron microscopy, the gold probes were silver intensified. Light microscopy demonstrated the absence of B-50 immunostaining in living neurons and the presence after permeabilization by fixation and subsequent treatment of the neurons with sodium borohydride, indicating that B-50 is located intracellularly. Both immunofluorescence and immunogold-silver labelling revealed that B-50 immunoreactivity outlined all neurites of the morphologically polarized neurons. For quantitative electron microscopy, six morphologically polarized neurons (developmental stage 3) were carefully selected from immunolabelled Epon-embedded neurons and processed completely to ultrathin sections. In this way the ultrastructural localization of B-50 has been studied in the cell body, the neurites and their growth cones. For each sectioned neuron, the relative distribution of the gold-silver deposits (representing B-50) over the plasma membrane of various cellular compartments was quantitated. B-50 is located at the plasma membrane of the neuronal cell body and all neurites including their growth cones. The density of B-50 on the plasma membrane of growth cones is not different from that of the neuritic shaft. In addition, B-50 is present on the cytosolic side of the membrane of small electron-lucent vesicles (average diameter 102.7 +/- 2.5 nm) resembling transport vesicles. These vesicles are present in the cell body and the neurites. A two-fold concentration is found in the central region of the growth cones, suggesting a role of these vesicles in axonal transport, membrane insertion and (or) recycling. Since, at the onset of neuronal polarization, B-50 is present at the plasma membrane in all compartments of the hippocampal neuron, we suggest that at this stage of development B-50 does not participate directly in the processes leading to morphological polarization.(ABSTRACT TRUNCATED AT 400 WORDS)     

Up-regulation of GAP-43 and growth of axons in rat spinal cord after compression injury

Summary The growth-associated protein-43 (GAP-43) is an axonal phosphoprotein which is expressed at high levels during development and is reinduced by regeneration in the PNS. Consequently it is believed to be a key molecule in the regulation of axonal growth. However, injury to the CNS does not result in significant regeneration and this has been suggested to correlate with a failure of central neurons to up-regulate GAP-43 after axotomy. We have examined a model of spinal cord injury which is unique in two respects; first dural integrity is maintained by compression of the cord with smooth forceps (thus excluding connective tissue elements) and, secondly, considerable axonal growth has been reported through the resulting lesion. Our previous studies have shown that GAP-43 is extensively distributed in the rat spinal cord (see accompanying paper), but here we have used anti-GAP-43 antiserum at a dilution which did not yield any immunostaining in normal cord. However, supranormal levels of GAP-43 were detected in cell bodies and axons around the lesion within four days of compression injury. Double immunostaining with the RT97 monoclonal antibody indicated that a small subpopulation of neurons local to the site of compression were axotomized and expressed GAP-43 and phosphorylated neurofilament epitopes in their cell bodies. Although damage to long axon tracts was extensive, there was no evidence of regeneration in white matter. On the other hand cavities which formed in grey matter provided an environment for axonal elongation. Immunolabelling with markers for astrocytes and endothelial cells was used to evaluate the interaction of elongating axons with endogenous CNS cell types. Sprouting axons, identified by the presence of elevated levels of GAP-43, did not appear to grow in contact with astrocytes but preliminary evidence suggested that newly formed capillaries provided an appropriate substrate.     

GAP-43治疗大鼠完全性脊髓损伤后GFAP表达的变化及意义

目的:通过制备完全性脊髓损伤(SCI)成年SD大鼠模型,研究生长相关蛋白(GAP-43)治疗大鼠SCI后胶质原纤维酸性蛋白(GFAP)的变化,探讨GAP-43在再生修复中的作用,为临床治疗提供实验参考。方法:咬除T7-T8棘突及相应椎板,用剪刀将脊髓完全横断,制成SCI模型。雌性8周龄SD大鼠75只,随机分为三组:GAP-43抗体组、GAP-43抗原组、对照组,每组25只。使用直接注射法将GAP-43抗原和GAP-43多克隆抗体分别注入抗原组和抗体组的大鼠脊髓的断端,观察各组大鼠肢体功能的恢复情况,用BBB评分法进行不同时段的行为学评分、免疫组化染色及图像分析方法观察GFAP的表达变化,并对其进行相关性分析。结果:对照组大鼠在不同时间段的行为学评分最低,抗原组评分最高;抗原组GFAP阳性细胞显著增多,而抗体组晚期则显著减少。结论:GAP-43可促进星形胶质细胞增生,而GAP-43抗体对星形胶质细胞的增生则表现为抑制作用。本实验结果表明GAP-43对脊髓损伤具有较好的治疗作用。     

Reciprocal age-related changes in GAP-43/B-50, substance P and calcitonin gene-related peptide (CGRP) expression in rat primary sensory neurones and their terminals in the dorsal horn of the spinal cord and subintima of the knee synovium

Age-related changes in the expression of the growth associated protein GAP-43/B-50, and the neuropeptides substance P and calcitonin gene-related peptide (CGRP) were investigated in the sensory neurones of rat dorsal root ganglia, dorsal horns of the spinal cord and subintimal knee synovium. The two time-points studied were 2 months (young adults) and 14-month (aged)-old Sprague Dawley rats. Dorsal root ganglia: In young adults, 40 and 35% of the L4-L5 dorsal root ganglion neurones were positive for GAP-43/B-50 with a 1.5 fold increase in frequency in aged rats at the L5 ganglion. GAP-43/B-50 was strongly expressed by the non-neuronal satellite cells of some medium and many large sized neurones in aged rats. There were marked reciprocal shifts between small and medium sized sensory neurones in respect to their substance P and CGRP expression profiles. Dorsal horn of the spinal cord: there was a 1.3 fold decrease of substance P at L5 level and a 1.3 and 1.5 fold decrease of CGRP at L4-L5 levels in aged rats, respectively. Synovial membrane: There was a 2.3 fold increase in GAP-43/B-50 and a 2.5 fold decrease of CGRP with no changes in substance P expression. These results indicate that (i) primary sensory neurones undergo age-related changes already in early stages of aging, (ii) aging may result in a reduction of substance P and CGRP axonal transport, and (iii) reduced numbers of CGRP containing synovial perivascular fibres may imply a deficient regulation of the synovial microvasculature and therefore metabolic homeostasis of the joint in aged subjects.     

臂丛损伤脊髓运动神经元与神经根GAP-43 mRNA表达

目的：探讨臂丛根性撕脱伤后脊髓腹角运动神经元胞体及其神经根GAP-43 mRNA的表达变化及其影响因素,为臂丛损伤的修复治疗提供理论依据.方法：本实验创立三种臂丛根性撕脱伤模型：C7前根撕脱（Ⅰ组）;C7前根撕脱＋切断同侧C5～T1后根（Ⅱ组）;C7前根撕脱＋C5和C6之间作同侧脊髓半横断（Ⅲ组）.术后2周按CBS评分标准检查动物神经缺失症状,用SYBR Green荧光定量RT-PCR方法检测脊髓腹角运动神经元胞体及其神经根GAP-43 mRNA的表达改变.结果：根据CBS评分标准,对照组计为0分,Ⅰ组计分较低、Ⅲ组计分最高.对照组C7神经元胞体和C7神经根中GAP-43 mRNA表达量相近,但三种损伤组术后2周神经元胞体内GAP-43 mRNA表达均上调,而神经根内表达却下调.结论：（1）臂丛根性撕脱伤后脊髓腹角运动神经元胞体GAP-43 mRNA表达受突触前机制的调控;（2）臂丛损伤2周时神经元胞体内GAP-43 mRNA表达呈现高峰期,此时进行神经移位术将显著提高神经修复的效果.     

GAP-43治疗大鼠脊髓横断后神经中丝NF200表达的变化

通过制备成年SD大鼠完全性脊髓损伤模型,研究生长相关蛋白(GAP-43)治疗大鼠脊髓损伤后神经中丝(NF200)表达的变化,探讨GAP-43在再生修复中的作用,为临床治疗提供实验依据。实验采用雌性8周龄SD大鼠75只,制成脊髓损伤模型后随机分为三组:GAP-43抗体组、GAP-43抗原组和对照组,每组25只。使用直接注射法将GAP-43抗原和GAP-43多克隆抗体分别注入抗原组和抗体组的大鼠脊髓的断端,对照组仅切断脊髓而不给药,最后观察各组大鼠肢体功能的恢复情况。用BBB评分法对不同时段大鼠的行为学表现进行评分,用HE染色及免疫组化染色观察NF200的表达,并对其进行相关性分析。结果发现对照组在不同的时间段行为学评分最低和抗原组评分最高,脊髓损伤区病理改变明显好转,NF200的表达呈进行性增高,且前角神经元NF200的表达早于后角神经元。抗体组早期恢复出现明显的停滞状态,但停药后能很快恢复。说明GAP-43能促进损伤脊髓的恢复,而抗体对损伤脊髓恢复的影响是可逆的,这对于脊髓再生的研究是一种值得探讨的新方法,对进一步探索脊髓损伤的治疗具有重要的意义。