重组睫状神经营养因子对受损周围神经施万细胞相关基因表达的作用

目的研究重组睫状神经营养因子(CNTF)对受损周围神经施万细胞基因表达的作用。方法用硅管套接切断的大鼠坐骨神经，在受损神经局部给予重组CNTF，术后用免疫组织化学ABC法结合计算机图像分析观测S100蛋白(S100)、生长相关蛋白-43(GAP-43)、磷酸化酪氨酸(PTyr)、信号转导子和转录激活子(STAT)3的免疫反应阳性物质在修复侧远段神经的分布和相对含量。结果CNTF组修复侧远段神经相应区域S100、GAP-43、PTyr、STAT3阳性物质的含量显著或非常显著高于生理盐水组。结论重组CNTF能上调受损神经施万细胞S100、GAP-43、PTyr和STAT3的表达，提示重组CNTF通过强化受损神经施万细胞JAK-STAT途径，E调其S100和GAP-43的基因表达。     

信号转导子与转录激活子3和细胞因子信号转导抑制因子-3蛋白在成年大鼠脊髓内的表达与定位

目的分析JAK-STAT信号转导途径重要成员STAT3蛋白和该途径抑制蛋白SOCS-3在大鼠脊髓内的表达和细胞内定位分布。方法免疫细胞化学技术和形态计量学方法。结果STAT3免疫阳性产物主要分布于脊髓前角运动神经元的胞浆内；SOCS-3免疫阳性产物广泛分布于脊髓前角及后角神经元内、部分胶质细胞及神经纤维内。在前角运动神经元内，SOCS-3免疫阳性产物主要分布于核内，有时也可分布于胞浆中。结论STAT3和SOCS-3广泛表达于正常大鼠脊髓内，其中SOCS-3具有核蛋白或胞浆蛋白两种存在形式。     

大鼠坐骨神经及其胞体信号传导子和转录激活子3和磷酸化酪氨酸的表达

目的：研究脊神经及其胞体中信号传导子和转录激活子３（ＳＴＡＴ３）的表达及其活化程度。方法；用免疫组化ＡＢＣ法显示成年大鼠坐骨神经，Ｌ４－５段脊髓和Ｌ５脊髓神经节中ＳＴＡＴ３和磷酸化酪氨酸，结果；坐骨神经轴突，脊髓前角外侧核神经元，脊神经节神经元胞体中ＳＴＡＴ３的含理较多。磷酸化酪氨酸的含昨很少。结论；脊神经及其胞体中存在一类酪氨酸激酶－信号传导子和转录激活子途径，但活化程度较低。     

CNTF及其受体在大鼠脊髓组织中的表达

CNTF具有广泛的神经营养活性 ,可能在神经发育与再生修复中发挥重要作用。有关 CNTF在大鼠脊髓组织中的表达尚不清楚 ,我们在制备抗人 CNTF多克隆抗体的基础上 ,利用免疫组化及原位杂交的方法系统的研究了成年大鼠脊髓组织 CNTF与受体 CNTFRα的分布与细胞定位。免疫组化结果显示 :CNTF免疫反应物质存在于脊髓灰质各层神经元中 ,包括运动神经元、中间灰质神经元及部分背角小细胞 ,尤其腹角运动神经元染色明显。胞浆胞核均有染色 ,胞核染色更深 ,无核仁染色 ,灰白质中有散在胶质细胞染色。原位杂交结果表明 ,CNTF m RNA的分布与…     

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。     

一氧化氮合酶阳性神经元及纤维在大鼠脊髓的分布及其来源

用还原型尼克酰胺腺嘌呤二核苷磷酸(NADPH)脱氢酶反应观察了大鼠脊髓内一氧化氮合酶阳性神经元和纤维的分布。胞体浓染且突起显示良好的类似Golgi染色的阳性神经元分布于亘脊髓全长的中央管周围灰质(X层),胸腰髓(T1-L3)的中间带外侧核、中介核及腰骶髓(L6、S1)的骶髓副交感核和后连合核,少量弥散于后角Ⅳ～Ⅵ层。后角Ⅲ层内可见较多的胞体浓染或淡染但突起短或未见阳性突起的小型神经元。前角内无阳性神经元,但有较多串珠状阳性终末支,并在运动神经元胞体和树突上形成终扣。阳性纤维和终末在后角Ⅰ层和Ⅲ层密集,其它部位稀少。在胸腰段和腰骶段脊髓后角内、外侧缘有行向腹侧的内、外侧阳性纤维束,外侧束集中而粗大,阳性纤维束终止于中间带外侧核和后连合核。切断后根和半横断颈髓的实验表明,后角Ⅰ层的阳性纤维和终末主要来源于脊神经节内一氧化氮合酶阳性细胞的中枢突,而Ⅲ层内的阳性纤维和终末则为Ⅲ层和后角深层阳性细胞的突起。结合文献和本文结果可以推断,一氧化氮在躯体和内脏感觉的传入过程及内脏传出活动中是一种重要的活性物质。     

P物质在成年猫脊髓和背根节的分布

目的 观察P物质在成年猫脊髓和背根节的表达情况。方法 成年健康雄猫5只，取L5脊髓节和随机取一侧L6背根节制作厚20μm冰冻切片，用免疫组织化学ABC法染色，观察SP在正常脊髓和背根节的分布。结果 在脊髓，SP阳性膨体主要见于后角Ⅰ、Ⅱ板层，后角Ⅳ、Ⅴ、Ⅵ板层及侧角的一些神经元呈弱阳性反应，前角运动神经元胞体和突起呈阳性反应，胞核不着色。在背根节，SP的免疫阳性物主要分布于小细胞的胞浆内，胞核不着色。结论 SP在成年猫脊髓和背根节有表达。     

人脊髓内NF样阳性神经元的形态及分布

目的：观察不同胎龄胎儿脊髓神经丝蛋白(NF)阳性神经元的形态、分布和发育变化，为脊髓-脊髓移植选择适宜的胎龄提供形态学依据。方法：胎儿脊髓19例，SP免疫组化染色，图像分析。结果：脊髓侧角内NF阳性神经元由中央管向外迁移；前角神经元由外向内迁移。NF阳性神经元密度在胚胎早期逐渐升高，晚期呈下降趋势。NF阳性神经元在胎龄16周时，胞体呈圆形、卵圆形，突起少沙，胞核大，有偏极现象，至32周时胞体呈锥形、梭形、多角形；胞体逐渐增大，胞浆逐渐增多，胞核多位居中央；突起增多。结论：未发育成熟的神经元内也有NF的存在。脊髓内NF阳性神经元密度随胎龄增加而逐渐增加，形态逐渐成熟，提示人脊髓-脊髓移植时以16周胎龄作供体较为适宜。     

大鼠脊髓神经肽Y、神经降压肽免疫反应神经元向结合臂旁核的投射

用HRP逆行束路追踪法结合免疫细胞化学双标记技术,研究了大鼠颈、胸、腰、骶各段脊髓神经肽Y(NPY)、神经降压肽(NT)免疫反应神经元向结合臂旁核的投射。HRP逆行标记细胞分布于两侧脊髓灰质第Ⅰ、Ⅱ、Ⅳ、Ⅴ、Ⅶ层、中央管背侧灰质连合及外侧颈核、外侧脊髓核中,以对侧为主。NPY免疫反应阳性神经元分布于两侧后角第Ⅰ层、第Ⅱ层浅部、外侧颈核、外侧脊髓核及骶髓后连合核中。NT免疫阳性神经元分布于两侧后角第Ⅰ层及第Ⅱ层中。在后角第Ⅰ层、外侧颈核和外侧脊髓核中,可观察到部分NPY-HRP双标记神经元;在后角第Ⅰ层可观察到部分NT-HRP双标记神经元。本研究结果提示,NPY和NT神经元可能参与脊髓-结合臂旁核的痛觉传递。     

周围神经损伤后局部一次给予重组CNTF对神经元变性和再生的作用

目的 :研究周围神经损伤后 ,局部一次给予重组CNTF对神经元变性和再生的作用。方法 :用硅管套接切断的成年大鼠坐骨神经 ,术时在损伤神经局部一次性给予重组CNTF ;用甲苯胺蓝染色和胆碱酯酶组织化学方法研究神经元的逆行变性 ,用HRP逆行追踪研究轴突的再生 ,并进行后足功能测试。结果 :重组CNTF减轻损伤运动神经元胞核偏位和胆碱酯酶活性的变化 ,增加轴突再生的运动神经元数量 ,改善功能指数。结论 :局部一次给予重组CNTF和再加皮下注射一样能保护损伤运动神经元并促其轴突再生。     

哮喘豚鼠初级传入神经元ERK的表达及NGF的调节作用

目的　探讨细胞外信号调节蛋白激酶 (ERK)在哮喘豚鼠初级传入神经元 (C7 T5脊神经节 )的表达及神经生长因子 (NGF)对ERK的调节作用。方法　利用免疫组织化学结合显微图像分析 ,研究哮喘豚鼠C7 T5脊神经节活化的ERK免疫反应变化。结果　与对照组相比 ,哮喘组豚鼠活化ERK免疫反应在C7 T5脊神经节神经元核内明显上调 (P <0 .0 1) ,其阳性细胞比率明显高于对照组。Anti NGF组豚鼠活化ERK免疫反应在C7 T5脊神经节神经元的核中明显低于哮喘组 (P <0 .0 1)。结论　C7 T5脊神经节神经元活化的ERK可能参与哮喘的发病过程 ,NGF可上调哮喘豚鼠C7 T5脊神经节神经元活化的ERK表达     

Analysis of the prolongation of rat eosinophil survival induced by recombinant rat interleukin-5

Rat eosinophil survival was prolonged by recombinant rat IL-5 prepared by the baculovirus expression system. The IL-5-induced prolongation of eosinophil survival was dose-dependently inhibited by the protein synthesis inhibitor cycloheximide, the DNA-dependent RNA synthesis inhibitor actinomycin D, and the tyrosine kinase inhibitor herbimycin A. The MEK-1 inhibitor PD98059 inhibited IL-5-induced phosphorylation of both p44 and p42 MAP kinases, but the IL-5-induced prolongation of eosinophil survival was not inhibited. In contrast, the JAK2 inhibitor AG490 inhibited the IL-5-induced prolongation of eosinophil survival. Treatment of eosinophils with IL-5 resulted in phosphorylation of STAT5 but not STAT1, and the IL-5-induced phosphorylation of STAT5 was inhibited by AG490. These findings suggest that recombinant rat IL-5 activates JAK2 tyrosine kinase, which phosphorylates STAT5, and induces protein synthesis required for the prolongation of rat eosinophil survival.     

Nogo(N-18)在成年大鼠三叉神经节、背根节及腰段脊髓中分布的免疫组织化学研究

目的：观察Nogo在成年大鼠三叉神经节、背根节及腰段脊髓中的分布，探讨其存在的意义。方法：采用抗Nogo(N-18)多克隆抗体免疫组织化学SP法。结果：成年大鼠三叉神经节、背根节感觉神经元胞浆和胞核以及腰段脊髓灰质神经元胞核内出现Nogo免疫反应。结论：Nogo免疫反应存在于大鼠三叉神经节、背根节感觉神经元胞浆和胞核以及腰段脊髓灰质神经元胞核内，应重新认识Nogo的功能。     

Locus coeruleus neurons in the rat containing neuropeptide Y, tyrosine hydroxylase or galanin and their efferent projections to the spinal cord, cerebral cortex and hypothalamus

The efferent projections of locus coeruleus neurons which contain neuropeptide Y-, tyrosine hydroxylase- or galanin-like immunoreactivity were investigated using the indirect immunofluorescence technique combined with the retrograde transport of the fluorescent substance Fast Blue. Four groups of rats received injections of Fast Blue: (1) bilaterally into the mid-thoracic spinal cord (T6-T7); (2) unilaterally into the low cervical spinal cord (C4-C5); (3) unilaterally into the paraventricular, periventricular and dorsomedial hypothalamic nuclei; and (4) unilaterally into five sites in the cerebral cortex (frontal, cingulate and striate cortex). Efferent projections to the spinal cord, hypothalamus and cerebral cortex from neuropeptide Y-, tyrosine hydroxylase- and galanin-containing locus coeruleus cells were observed. A higher percentage of the peptidergic locus coeruleus neurons projected to the hypothalamus than to the spinal cord or cerebral cortex. The distribution and morphology of the neuropeptide Y- and galanin-containing neurons in the locus coeruleus were also investigated. Neuropeptide Y-like immunoreactivity and galanin-like immunoreactivity were found in small, medium and large multipolar neurons, as well as in fusiform locus coeruleus cells. The neuropeptide Y- and galanin-immunoreactive neurons were found throughout the locus coeruleus. In the caudal locus coeruleus, they were primarily located in the dorsal portion. Neuropeptide Y-like immunoreactivity and galanin-like immunoreactivity were only seen in a few tyrosine hydroxylase-positive neurons of the subcoeruleus group. The data show that the peptide-containing locus coeruleus neurons have efferent projections to the spinal cord, hypothalamus and cerebral cortex. The locus coeruleus may be divided into functional subdivisions dependent on the region of the locus coeruleus, the neurotransmitter/neuropeptide(s) contained within the neurons and their efferent projections.     

Critical proline residues of the cytoplasmic domain of the IL-5 receptor {alpha} chain and its function in IL-5-mediated activation of JAK kinase and STAT5

The high-affinity receptor (R) for IL-5 consists of a uniquea chain (IL-5Rc) and a ß chain (ßc) thatisshared with the receptors for IL-3 and granulocyte macrophagecolony stimulating factor (GM-CSF). We defined two regions ofIL-5R for the IL-5-induced proliferative response, the expressionof nuclear proto-oncogenes, and the tyrosine phosphorylationof cellular proteins including ßc, SH2/SH3-containingproteins and JAK2 kinase. In the studies described here, wedemonstrate that IL-5, IL-3 or GM-CSF stimulation induces thetyrosine phosphorylation of JAK2, and to a lesser extent JAK1,and of STAT5. Mutational analysis revealed that one of the prolineresidues, particularly Pro352and Pro355, in the membrane-proximalproline-rich sequence (Pro352-Pro353-X-Pro355) of the cytoplasmicdomain of IL-5R is required for cell proliferation, and forboth JAK1 and JAK2 activation. In addition, transfectants expressingchimeric receptors which consist of the extracellular domainof IL-5R and the cytoplasmic domain of ßc respondedtoIL-5 for proliferation and tyrosine phosphorylation of JAK1.Intriguingly, electrophoretic mobility shift assay analysisrevealed that STAT5 was activated in cells showing either JAK1or JAK2 tyrosine phosphorylation. These results indicate thatactivation of JAK1, JAK2 and STAT5 is critical to coupling IL-5-inducedtyrosine phosphorylation and ultimately mitogenesis, and thatPro352 and Pro355 in the proline-rich sequence appear to playmore essential roles in cell growth andin both JAK1/STAT5 andJAK2/STAT5 activation than Pro353 does.