糖尿病大鼠脊髓nNOS免疫阳性神经元的变化

目的：观察糖尿病大鼠脊髓神经元型一氧化氮合酶(nNOS)免疫阳性神经元数量的变化，探讨NO在糖尿病发生和发展中的作用机制。方法：用链脲佐菌素诱导建立糖尿病大鼠模型，ABC免疫细胞化学法显示nNOS免疫阳性神经元。结果：大鼠脊髓内nNOS阳性神经元主要分布于中央管周围灰质和中间带等区域。中间带外侧核可见nNOS免疫阳性神经元较集中，细胞突起呈束状伸向中央管周围灰质方向；定量分析显示，糖尿病大鼠脊髓中央管周围灰质和中间外侧核在7w、12w时nNOS免疫阳性神经元数量明显增多。结论：糖尿病时伤害性刺激的传人增多，增多的nNC)S免疫阳性神经元可能与痛觉过敏等糖尿病周围神经病变有关。     

成年弥猴背根神经节nNOS免疫阳性神经元的分布

目的：观察正常成年称猴背根神经节神经元型一氧化氮合酶(nNOS)免疫阳性神经元的分布。方法：ABC免疫细胞化学方法显示nNOS免疫阳性神经元，并用体视方法进行定量分析。结果：猕猴颈、胸、腰各段背根神经节nNOS免疫阳性神经元的分布相似，数量较多，阳性神经元的大小不等，多呈圆形或椭圆形；胞浆着色较深，胞核位于细胞中央，不着色，细胞被神经纤维束分隔成群。nNOS免疫阳性神经元以中型神经元为主，其次为小型神经元，其胞浆呈强阳性染色，细胞直径＜50μm，大型神经元较少。颈、胸、腰各段背根神经节nNOS免疫阳性神经元的密度以及阳性细胞与总细胞数的比值均无明显差异。结论：称猴背根神经节nNOS主要表达在中、小型神经元，提示NO可能主要参与痛觉等浅感觉的传导和调制。     

Nestin免疫阳性结构在正常成年大鼠脑内的分布及形态学观察

目的观察成年大鼠脑内Nestin免疫阳性结构的分布和形态学特征。方法采用免疫组织化学的方法 (ABC法)对成年SD大鼠脑切片进行Nestin免疫组化染色。结果在成年大鼠脑内基底前脑的隔斜角带复合体、Meynert基底核、视上核、室管膜区、室管膜下区、海马齿状回、CA1区、CA3区、正中隆起、穹窿下器、最后区处有Nestin免疫阳性结构的表达,形态有3种类型。结论成年大鼠脑内多处存在Nestin阳性结构的表达,形态多样,其化学属性和生物学意义有待进一步研究。     

正常成年大鼠神经元前体细胞的迁移

目的探讨正常成年大鼠脑中神经元前体细胞的迁移路径和迁移特征,为病理状况下的神经元迁移研究提供理论和实验基础。方法将成年大鼠脑组织按冠状位和矢状位行冰冻切片（片厚20μm）,采用免疫组织化学染色技术观察Doublecortin（DCX）免疫阳性细胞的迁移路径以及DCX阳性细胞的迁移特征。结果正常成年大鼠神经元前体细胞的迁移主要有两条路径：一条是从室下区到嗅球的吻侧迁移流,另一条是位于皮层和胼胝体之间的胼胝体周围迁移流。吻侧迁移流中的DCX免疫阳性细胞较规则的有序排列,形态上呈梭形的胞体和单个较长的前导突起,前导突起基本朝向嗅球,细胞大小基本一致：而胼胝体周围迁移流中的DCX免疫阳性细胞胞体呈类圆形,突起的大小、数目和方向多样。结论研究神经元前体细胞的迁移不仅有助于明确其迁移的具体机制,而且有助于控制神经元的迁移和设计有效的治疗策略。     

脑红蛋白在成年大鼠脊髓中的分布

目的观察脑红蛋白（NGB）在成年大鼠脊髓中的分布。方法制备成年大鼠脊髓的冰冻切片,用免疫组织化学染色方法研究NGB蛋白在成年大鼠脊髓内的分布及细胞定位。结果在脊髓的颈、胸、腰段,NGB免疫反应阳性细胞主要分布于脊髓灰质中,其范围包括灰质脊髓前角、中间带和后角。NGB免疫反应阳性物质广泛存在于脊髓灰质神经元的胞浆中。结论NGB蛋白在大鼠脊髓灰质有广泛的表达,提示NGB在脊髓功能活动中可能起重要作用。     

糖尿病大鼠中脑导水管周围灰质nNOS免疫阳性神经元的变化

糖尿病患者常并发慢性周围神经病变,表现为感觉异常。一氧化氮(NO)缺乏导致周围神经供血不足以及由此引起的感觉传导异常可能与之有关。但对于中枢神经是否参与感觉障碍?与糖尿病时感觉传导异常的关系如何,仍有许多未知。中脑导水管周围灰质(PAG)是内源性镇痛系统的关键结构,其背外侧区富含神经元型一     

表皮生长因子在成年猴脊髓的表达

目的探讨EGF(epidermal growth factor)在成年猴脊髓的表达。方法EGF特异性抗体的免疫细胞化学染色技术(SP法)。结果EGF免疫阳性物质主要位于神经元胞浆内，前角前内侧核、前角前核和前角前外侧核的大运动神经元以及背核的大神经元具有强免疫阳性信号，中等强度免疫阳性神经元主要分布于前角后内侧核、前角中央核、前角后外侧核和前角后外侧后核、中间内侧核和中央管周围，弱阳性神经元主要位于后角边缘核、胶状质、后角固有核和后角连合核。此外，在脊髓白质中可见部分EGF阳性胶质细胞和纤维。结论EGF免疫阳性反应产物在猴脊髓有广泛的分布，提示其功能可能涉及多种神经元和非神经细胞，为探讨表皮生长因子在猴脊髓分布规律和功能提供了有价值的形态学资料。     

成年大鼠脊髓背角神经元受损轴突在外周神经移植物中的再生速度

用抗神经丝(anti-neurofilament)单克隆抗体免疫组织化学技术,研究大鼠脊髓背角神经元受损轴突在外周神经移植物中的再生速度,发现脊髓背角神经元受损轴突在外周神经移植物中的最短初始延搁时间为4d;其后在一定时期内不断有再生神经纤维长入移植物,即不同神经纤维初始延搁时间不一致;再生轴突在移植物中的生长速度也不一致,最快再生速度为2.14mm/d。     

大白鼠脊髓前角内含γ—氨基丁酸神经元的化学神经解剖学研究

用ＨＲＰ与免疫细胞化学结合法和免疫电镜方法观察了大白鼠脊髓Ｌ４～５节段内前角含γ－氨基丁酸（ＧＡＢＡ）神经元的分布及其与躯体传出的关系。光镜下，在脊髓前角各层，包括位于前角的前外侧部的Ｒｅｘｅｄ ＩＸ层，均有ＧＡＢＡ免疫反应阳性神经元胞体和梢分布。ＧＡＢＡ阳性胞体为圆形或三角形，具有多个突起，可分大、中两型。电镜下，ＧＡＢＡ样免疫反应产物呈细小颗粒状沉淀，分布于核周质、树突和轴突内。在轴突末梢，免     

蛋白激酶Cγ在成年大鼠延髓和脊髓白质中的分布

目的 观察蛋白激酶Cγ亚型(PKCγ)在成年大鼠延髓和脊髓白质内的分布.方法 免疫组织化学ABC法显示PKCγ免疫阳性物质在延髓和脊髓颈、胸、腰、骶段白质内的表达和分布.结果 PKCγ免疫阳性物质分布于锥体、锥体交叉及脊髓颈、胸、腰、骶段后索腹侧中;延髓和脊髓背角神经元中也有PKCγ免疫阳性物质存在.结论 PKCγ免疫阳性反应产物广泛分布于成年大鼠延髓和脊髓白质内的皮质脊髓束中.PKCγ免疫阳性反应产物在延髓锥体和脊髓后索内的定位与大鼠皮质脊髓束的位置和走行一致,提示其在运动神经传导通路中可能起着重要作用.     

正常成年SD大鼠脑内CXCR4免疫阳性细胞的形态学观察

目的 研究成年大鼠脑内表达CXCR4细胞的形态学特征.方法 用免疫组织细胞化学方法对成年大鼠脑内表达CXCR4细胞的分布和形态特点.结果 在成年大鼠脑内,CXCR4免疫阳性细胞广泛分布于新皮质各层和各区.在皮层上,外颗粒层阳性细胞相对比较集中,形成一个深染的阳性细胞带,而其它各层阳性细胞比较分散.在各区皮质,如压部皮质...     

Electrophysiological characteristics of identified kidney-related neurons in adult rat spinal cord slices

Whole-cell patch-clamp recordings were made from kidney-related neurons in the intermediolateral cell column (IML) in horizontal slices of thoracolumbar spinal cord from adult rats. Kidney-related neurons were identified in vitro subsequent to inoculation of the kidney with a fluorescent, retrograde, transynaptic pseudorabies viral label (i.e., PRV-152). Kidney-related neurons detected in the IML expressed choline acetyltransferase, characteristic of spinal preganglionic motor neurons. Their mean resting potential was −51 ± 4 mV and input resistance was 448 ± 39 MΩ. Both spontaneous inhibitory and excitatory post-synaptic currents (i.e., sIPSCs and sEPSCs) were observed in all neurons. The mean frequency for sEPSCs (3.1 ± 1 Hz) was approximately 2.5 times that for sIPSCs (1.4 ± 0.3 Hz). Application of the glycine and GABA_A receptor-linked Cl⁻ channel blocker, picrotoxin (100 μM) blocked sIPSCs, while the ionotropic glutamate receptor antagonist, kynurenic acid (1 mM) blocked all sEPSCs, indicating they were mediated by GABA/glycine and glutamate receptors, respectively. Thus, using PRV-152 labeling allowed whole-cell patch-clamp recording of neurons in the adult spinal cord, which were kidney-related. Excitatory glutamatergic input dominated synaptic responses in these cells, the membrane characteristics of which resembled those of immature IML neurons. Combined PRV-152 pre-labeling and whole-cell patch-clamp recordings may allow more effective analysis of synaptic plasticity seen in adult models of injury or chronic disease.     

Auto-catalytic ceria nanoparticles offer neuroprotection to adult rat spinal cord neurons

This paper describes the evaluation of the auto-catalytic anti-oxidant behavior and biocompatibility of cerium oxide nanoparticles for applications in spinal cord repair and other diseases of the central nervous system. The application of a single dose of nano-ceria at a nano-molar concentration is biocompatible, regenerative and provides a significant neuroprotective effect on adult rat spinal cord neurons. Retention of neuronal function is demonstrated from electrophysiological recordings and the possibility of its application to prevent ischemic insult is suggested from an oxidative injury assay. A mechanism is proposed to explain the auto-catalytic properties of these nanoparticles.     

低水平铅暴露大鼠齿状回nNOS阳性神经元的时程变化

目的:了解齿状回nNOS阳性神经元在铅影响学习记忆神经毒机理中的作用。方法:采用免疫组织化学ABC法,观察了大鼠腹腔注射130mg/kg醋酸铅溶液染毒3h,6h,12h和24h后齿状回nNOS阳性神经元数目的变化。结果:与对照组相比,染铅12h时,齿状回nNOS阳性神经元数目均明显减少(P<0.01),染铅24h时,齿状回uNOS阳性神经元数目仍处于较低水平(P<0.05)。结论:本研究提示齿状回nNOS阳性神经元的时程变化可能为揭示铅对学习记忆影响的分子机制提供了一定的实验依据。     

大鼠系膜增生性肾小球肾炎模型的病理形态学观察

目的 :观察大鼠系膜增生性肾小球肾炎 (MsPGN)模型的病理形态学变化。方法 :采用一次性鼠尾静脉注射小鼠抗大鼠Thy1 1单克隆抗体法复制大鼠MsPGN模型 ,应用光镜、荧光显微镜、偏振光显微镜及透射电镜观察模型大鼠肾小球系膜细胞及系膜基质的变化 ;结果 :系膜细胞明显增生 ,细胞外基质 ,包括Ⅰ、Ⅲ胶原、纤维粘连蛋白 (FN)及层粘连蛋白 (LN)增多 ,个别大鼠肾小球出现硬化及纤维化。结论 :复制出具有典型病变的大鼠MsPGN模型。     

Neonatal bladder inflammation alters activity of adult rat spinal visceral nociceptive neurons

Purpose: This investigation examined the effect of inflammation produced by intravesical zymosan during the neonatal period on spinal dorsal horn neuronal responses to urinary bladder distension (UBD) as adults. Methods: Female rat pups (P14–P16) were treated with intravesical zymosan or with anesthesia-only. These groups of rats were subdivided forming four groups: half received intravesical zymosan as adults and half received anesthesia-only. One day later, rats were anesthetized, the spinal cord was transected at a cervical level and extracellular single-unit recordings of L6-S1 dorsal horn neurons were obtained. Neurons were classified as Type I—inhibited by heterotopic noxious conditioning stimuli (HNCS) or as Type II – not inhibited by HNCS – and were characterized for Spontaneous Activity and responses to graded UBD (20–60 mm Hg). Results: 227 spinal dorsal horn neurons excited by UBD were characterized. In rats treated as neonates with anesthesia-only, Type II neurons demonstrated increased spontaneous and UBD-evoked activity following adult intravesical zymosan treatment whereas Type I neurons demonstrated decreased spontaneous and UBD-evoked activity relative to controls. In rats treated as neonates with intravesical zymosan, the spontaneous and UBD-evoked activity of both Type I and Type II neurons increased following adult intravesical zymosan treatment relative to controls. Conclusions: Neonatal bladder inflammation alters subsequent effects of acute bladder inflammation on spinal dorsal horn neurons excited by UBD such that overall there is greater sensory neuron activation. This may explain the visceral hypersensitivity noted in this model system and suggest that impaired inhibitory systems may be responsible.     

Morphological and developmental characterization of local-circuit neurons in lamina III of the rat spinal cord

Using the Golgi silver impregnation technique the present study examines the morphology and development of presumptive local circuit neurons in lamina III of the rat lumbar spinal cord. These neurons generate local axonal plexuses which remain within the gray matter and dendritic trees which arborize in lamina III and the inner zone of lamina II. Analysis of developmental stages supports the contention that these neurons have local axons which do not enter the white matter. These cells undergo axonal and dendritic maturation during the postnatal period, well after the maturation of long axon neurons. This pattern parallels the development of local circuit neurons in lamina II.     

Somatostatin directly inhibits substantia gelatinosa neurons in adult rat spinal dorsal horn in vitro

Effects of somatostatin (SST) on the synaptic transmission to substantia gelatinosa (SG) neurons of adult spinal cord slices were investigated using intracellular recording and blind whole-cell patch-clamp technique. Bath application of SST (1 microM) induced the membrane hyperpolarization that was accompanied by a decrease in input resistance and had the reversal potential of -92 +/- 3 mV (n=5) in the intracellular recording experiment. In patch-clamp experiment, SST (1 microM) induced an outward current with amplitude of 14 +/- 2 pA (n=60) at the holding potential of -60 mV, and was not affected by TTX (n=3). The effect was dose-dependent with EC50 value of 0.82 microM (Hill coefficient: 0.89). The outward current was suppressed when the patch-pipette solution containing potassium channel blockers, Cs+ and tetraethylammonium (TEA), and was inhibited by Ba2+ (200 microM) to 15 +/- 6% of the control (n=3). In addition, the SST current reversed its polarity at potential close to the equilibrium potential of K+ channel calculated by the Nernst equation. No significant changes were found in amplitude and frequency of miniature excitatory postsynaptic currents (mEPSCs) and dorsal root evoked EPSC (eEPSC) by SST. Also, SST did not affect both of the miniature inhibitory postsynaptic currents (mIPSCs) and evoked inhibitory postsynaptic currents (eIPSCs), mediated by either the GABA or glycine receptor. We conclude that SST activates the K+ channel resulting in postsynaptic hyperpolarization in adult rat SG neurons without affecting presynaptic component of the transmission, which are considered to account, at least a part, for the analgesic effects of SST reported previously.     

急性给锂小鼠大脑皮层一氧化氮合酶阳性神经元的时程变化

目的：为阐明锂对脑发育影响的机理,探讨急性给锂不同时程对小鼠大脑皮层神经元型一氧化氮合酶(nNOS)阳性神经元的影响。方法：小鼠腹腔注射氯化锂,采用ABC免疫组织化学方法,观察急性给锂后24h内不同时程小鼠大脑皮层nNOS阳性神经元数目的变化。结果：急性给锂即刻小鼠大脑皮层nNOS阳性神经元数目明显增加,1h后达到高峰,6h和12h恢复到正常水平,24h较12h又有所增高,但仍处于正常水平。结论：急性给锂对小鼠大脑皮层nNOS活性有一定影响,这种变化可能是锂神经毒性的机制之一。     

癫痫大鼠海马结构谷氨酸和nNOS神经元的动态变化

的：探讨谷氨酸(Glu)和一氧化氮(NO)二者在癫痫模型中的作用及其相关性。方法：戊四唑化学点燃癫痫大鼠,分为Ⅰ、Ⅲ、Ⅴ级组和Ⅴ级后24h组。采用免疫组织化学方法和图像分析技术。结果：(1)Ⅲ、Ⅴ级组Glu免疫反应阳性神经元数目和平均光密度值升高;但Ⅴ级比Ⅲ级组有所下降;Ⅴ级后24h组恢复到对照组水平。(2)Ⅴ级组和Ⅴ级后24h组nNOS免疫反应阳性神经元数目和平均光密度值升高。结论：戊四唑点燃癫痫模型中,随着点燃级别的进展,Glu的含量是呈先增加后减少的趋势,而NO的含量是逐渐增加的,提示二者在癫痫发作中既有相关性又有独立性。