NMDAR1和GABA_A受体的α_1及α_3亚单位在成年猫小脑的定位分布

用免疫组织化学反应及 Nissl染色探讨了 NMDAR1及 GABAA受体α1 和α3亚单位在成年猫小脑皮质及小脑核的定位分布。结果表明 ,NMDAR1免疫反应产物主要分布在 Purkinje细胞胞质和分子层的树突 ,分子层的星形细胞和篮细胞以及颗粒细胞层的颗粒细胞和胶质细胞呈中等强度的阳性反应 ,小脑核的神经元胞质和部分突起着色明显。 GABAA受体的α1 亚单位免疫反应产物主要分布在 Purkinje细胞胞质和树突 ,分子层的星形细胞和胶质细胞呈弱阳性 ,小脑核的神经元阳性反应明显。GABAA 受体的α3亚单位免疫反应产物主要分布在 Purkinje细胞胞质和树突 ,分子层的星形细胞和篮细胞着色明显 ,胶质细胞呈免疫反应弱阳性 ,小脑核神经元及纤维着色明显。在 Purkinje细胞层 NMDAR1、GABAA受体α1 及α3亚单位免疫阳性神经元分别占 Purkinje细胞总数的 80 % ,61% ,88%。结论 :NMDAR1、GABAA受体的α1 及α3亚单位在成年猫小脑具有广泛的分布。这些受体在介导小脑的复杂功能中可能发挥重要的作用。     

NMDARl和GABAA受体的α1及α3亚单位在成年猫小脑的定位分布

用免疫组织化学反应及Nissl染色探讨了NMDARl及GABAA受体α1和α3亚单位在成年猫小脑皮质及小脑核的定位分布。结果表明，NMDARl免疫反应产物主要分布在Purkinje细胞胞质和分子层的树突，分子层的星形细胞和篮细胞以及颗粒细胞层的颗粒细胞和胶质细胞呈中等强度的阳性反应，小脑核的神经元胞质和部分突起着色明显。GABAA受体的α1亚单位免疫反应产物主要分布在Purkinje细胞胞质和树突，分子层的星形细胞和胶质细胞呈弱阳性，小脑核的神经元阳性反应明显。GABAA受体的α3亚单位免疫反应产物主要分布在Purkinje细胞胞质和树突，分子层的星形细胞和篮细胞着色明显，胶质细胞呈免疫反应弱阳性，小脑核神经元及纤维着色明显。在Purkinje细胞层NMDARl、GABAA受体α1及α3亚单位免疫阳性神经元分别占Purkinje细胞总数的80％，61％，88％。结论：NMDAR1、GABAA受体的α1及α3亚单位在成年猫小脑具有广泛的分布。这些受体在介导小脑的复杂功能中可能发挥重要的作用。     

脑干内5-HT能神经元至小脑皮质的纤维投射

本文用单纯免疫细胞化学和免疫电镜方法研究了大白鼠小脑皮质内5-HT纤维的分布及存在形式,并用HRP逆行追踪和免疫细胞化学结合法研究了脑干内5-HT能神经元向小脑皮质的纤维投射。实验证实:5-HT纤维广泛存在于小脑皮质各部、各层,其中Ⅴ～Ⅷ叶较集中,蚓部纤维密度较半球高。皮质各层中,颗粒细胞外层及Purkinje细胞层纤维密度最高,呈树枝状分布;分子层次之,呈“T”型分布;颗粒细胞内层纤维密度最低,向垂直皮质表面方向行走。电镜观察到:小脑皮质分子层内有多个5-HT免疫反应阳性末梢,其中多数为无突触末梢,部分分布在毛细血管壁附近,少数为轴-轴型突触末梢。实验还证实:小脑内5 HT纤维有多个来源,它们可来自延髓外侧网状核、中缝隐核、巨细胞网状核、楔外核、脑桥被盖网状核。     

猫小脑皮质结构的年龄相关性变化

目的：对青年和老年猫小脑皮质结构的年龄性变化进行比较。方法：动用Nissl染色显示小脑皮质神经元,免疫组织化学方法显示胶质纤维酸性蛋白免疫阳性（GFAP-IR）星形胶质细胞和神经丝蛋白免疫阳性（NF-IR）结构。显微镜下观察测量小脑皮质厚度和细胞密度。结果：与青年猫比较,老年猫小脑皮质总厚度及分子层厚度显著下降,颗粒层厚度明显增加,各层神经元密度明显降低;颗粒层中GFAP-IR细胞密度显著增加,阳性反应增强;老年猫蒲肯野细胞（PC）NF免疫阳性树突分支明显减少。结论：衰老过程中小脑皮质神经元丢失和PC中NF阳性树突减少,可能会导致老年小脑皮质接受和整合信息的功能降低,而星形胶质细胞活动增强对皮质神经元可能起保护作用。     

小鼠小脑皮质的组织发生

目的:探讨小鼠小脑皮质的组织发生过程。方法:应用光镜和电镜技术对胚胎和生后小脑皮质进行形态学观察,对各层厚度和细胞密度进行测量。结果:胚胎12 d(E12)小脑原基有室管膜层、套层和边缘层构成,约出生当日(P0)出现外颗粒层、分子层、Purkinje细胞层和内颗粒层。外颗粒层P6/7达最厚,至P20消失。P0至P30内颗粒层细胞逐步分化发育成熟,Purkinje细胞树突树逐渐形成,约P7时Purkinje细胞排列成单层。结论:E12至P0片层化小脑主要经历了细胞增殖、分化与迁移;P0至P30片层化结构逐渐发育成熟,外颗粒层消亡以细胞迁移和凋亡为主,其他各层细胞主要经历了分化发育与凋亡。     

一氧化氮合酶同功异构酶在大鼠中枢神经系统的分布

本文采用免疫组织化学技术观察三种一氧化氮合酶同功异构酶在正常大鼠中枢神经系统的分布。结果显示：ＮＯＳ１ 阳性神经元分布于中枢神经系统的广泛区域,包括大脑皮质Ⅱ、Ⅲ、Ⅵ层,海马分子层,齿状回多形层,尾壳核,中脑上丘,小脑皮质分子层和颗粒层,脊髓Ⅰ、Ⅱ层、侧角、中央灰质和前角。脊髓中央管室管膜上皮也为阳性。ＮＯＳ３ 阳性神经元的分布区域与前者有部分重叠,但也有差别：大脑皮质相同层次内的ＮＯＳ３ 阳性神经元的形态和染色不同于ＮＯＳ１;密集分布于海马锥体细胞层和齿状回颗粒细胞层;小脑浦肯野细胞ＮＯＳ３ 阳性;另外,ＮＯＳ３ 阳性神经元弥散分布于脊髓内,其胞体和核仁均呈阳性。ＮＯＳ２ 阴性。广泛的分布表明ＮＯ 与中枢神经系统的诸多功能有关,而ＮＯＳ１ 与ＮＯＳ３ 分布的差异又说明中枢神经系统不同神经元内ＮＯ的产生是由此两种同功异构酶分别参与而完成的。     

肝硬化后大鼠小脑皮质中P物质(SP)的变化及其意义-荧光免疫组化技术结合激光扫描共聚焦显微镜的研究

目的 :研究P物质 (SP)在大鼠小脑皮质中的分布特点及肝硬化后对大鼠小脑皮质中SP的影响。方法 :采用荧光免疫组化技术结合激光扫描共聚焦显微镜技术。结果 :①SP样阳性纤维及终末散在分布于小脑皮质各层 ,但在Purkinje神经元层其平均荧光强度相对其它各层较高 ,并主要分布于该神经元树突侧胞体周边形成突触样结构 ;②肝硬化后SP在小脑皮质中的分布与正常组基本一致 ,但阳性反应物的密集度明显增高 ,平均荧光强度显著增强。结论 :肝硬化后小脑皮质中SP含量较正常明显增多 ,很可能是脑损害的一种继发性代偿作用。     

Alzheimer病患者和大鼠脑内早老蛋白-1的分布

用免疫组化方法和原位杂交观察散发性阿尔茨海默病 (Sporadic Alzheimer' s disease)病人和老年对照 ,以及成年大鼠脑组织内的早老蛋白 -1(Presenilin-1,PS1)分布。结果观察到 PS1在脑内广泛分布 ,其中小脑的 Purkinje细胞、皮层的第 、 层细胞、内嗅皮层的第 层、海马的锥体细胞层和颗粒细胞层、黑质等结构 PS1表达较高。在大鼠和正常对照人脑内大多数 PS1阳性神经元染色呈点状分布于胞浆内 ;而阿尔茨海默病人脑内有许多 PS1阳性神经元为均质染色并且其中有些呈神经原纤维缠结状 ,其次有大量各型 PS1阳性老年斑 ,和少量 PS1阳性胶质细胞。结果提示 PS1可能参与散发性阿尔茨海默病脑病理改变     

小鼠小脑皮质发育中的神经元凋亡

目的探讨小鼠小脑皮质发育中神经元凋亡的规律和机制。方法用激活型Caspase-3多抗免疫组织化学标记及Hoechst 33258染色液染色,检测从出生至成年小鼠小脑皮质中神经元的凋亡,用Western blotting方法对小脑组织中Caspase-3和Caspase-8的活化片段进行半定量测定。结果外颗粒层、普肯耶细胞层和内颗粒层凋亡细胞密度最高分别在出生后第8d(P8)、P5及P9,P20各层凋亡细胞密度都很低。Caspase-3活化片段的表达量在P5较高,P5以后逐渐降低,至P14消失;Caspase-8活化片段的表达量从P0到P10都较高,P10以后逐渐降低,至P30消失。结论P0至P14是小脑皮质神经元凋亡的重要时期,通过启动Caspase-8的活化进而激活效应性Caspase-3的细胞凋亡途径存在于小脑皮质的塑型成熟过程。     

成年大鼠小脑Purkinje神经元不表达神经元特异性的钾离子氯离子共转运体2

目的:观察神经元特异性标记物钾离子氯离子共转运体2(KCC2)在成年大鼠小脑皮质的表达。方法:成年SD大鼠,4%多聚甲醛灌注固定,取脑,冷冻切片,KCC2和小脑Purkinje神经元标记物Calbindin-D28K在小脑的免疫荧光双标,激光共聚焦显微镜观察。结果:KCC2在颗粒细胞层神经元有丰富的表达,但在Purkinje神经元表达很弱。结论:神经特异性的KCC2可能不是成年大鼠的Purkinje神经元的离子型γ-氨基丁酸受体发挥抑制性作用的主要因素。     

大白鼠交感神经节内神经肽Y(NPY)样神经元的分布及其功能意义

本文用免疫细胞化学技术研究了大白鼠交感神经的颈上节,星状节,脑部和腹部的交感干节,腹腔节以及肠系膜上、下节内的NPY样免疫反应细胞体的分布。发现NPY样免疫反应细胞体为圆形或椭圆形。NPY样免疫反应产物呈颗粒状分布于细胞质中,多数交感神经节内的免疫反应细胞聚集成群,但椎前神经节内的NPY样免疫反应细胞较分散。本文并讨论了NPY存在于外周去甲肾上腺素能细胞中;NPY可以抑制由于电刺激引起的大白鼠输精管痉挛;局部动脉灌流可引起动脉收缩等问题。本文认为交感神经节中含有的NPY样免疫反应神经元与外周去甲肾上腺素能神经元为同一神经元。NPY对交感神经可能有兴奋或抑制功能。     

大白鼠甲状腺NPY样神经元的分布

应用免疫细胞化学PAP法,在光镜下观察了6只大白鼠甲状腺NPY样免疫反应神经元。甲状腺被膜与甲状腺实质之间有NPY样免疫反应神经元的胞体,一般沿被膜深面单行排列,但在甲状腺上极附近密集成群。胞体为椭圆形或圆形。甲状腺滤泡之间常见有三、五成群NPY样免疫反应神经元胞体形成的神经节。NPY样免疫反应纤维缠绕着血管壁或走行于甲状腺滤泡之间。本文对NPY样神经元与外周交感神经之间的关系进行了讨论。     

神经肽Y样免疫反应神经元在蟾蜍视网膜的定位与分布

用免疫组织化学ABC方法研究神经肽Y(NPY)样免疫反应神经元在蟾蜍视网膜的定位、形态与分布。结果表明:NPY样免疫反应仅存在于无长突细胞中。在102个免疫反应阳性无长突细胞中,92%为Ⅰ型,8%为Ⅱ型。NPY样免疫反应阳性纤维分布于内网层并密集呈三条带,分别位于第1亚层,第2、3亚层交界处和第4、5亚层之间。在视网膜平铺片上,NPY样免疫反应阳性胞体均匀分布于视网膜中央区和周围区,其密度为27±8个/mm~2(均数±标准差)。它们的树突野呈对称和非对称两型,前者的树突野大小为300—500μm×100—300μm,后者的长突约为200—400μm。     

胎儿(36—40周)纹状体和下丘脑NPY免疫反应神经元的分布

本文用免疫细胞化学ABC法研究胎儿(36—40周)纹状体和下丘脑NPY免疫反应神经元的分布。在胎儿的纹状体发现尾状核和壳含有许多NPY免疫反应神经元的胞体和纤维。这两个区域的一些血管壁内也含有NPY免疫反应神经元的胞体和纤维。胎儿下丘脑的漏斗核和视上核含密集的NPY免疫神经元胞体,在室旁核偶尔也可见到少数免疫神经元胞体。本文首次报道了尾状核和壳的血管壁内及视上核含有NPY免疫反应神经元胞体。     

Demonstration of a neuropeptide Y (NPY)-like immunoreactivity in the pigeon retina

The distribution of neuropeptide Y (NPY)-like immunoreactivity in the pigeon retina was investigated by fluorescence immunohistochemistry. NPY-positive cells were found in central and peripheral retina. NPY somata were located in the proximal portion of the inner nuclear layer and their processes directed to the inner plexiform layer where they ramified in 3 immunoreactive bands. NPY might play a role as a neurotransmitter or neuromodulator in the pigeon retina.     

Otolith organ or semicircular canal stimulation induces <Emphasis Type="Italic">c-fos</Emphasis> expression in unipolar brush cells and granule cells of cat and squirrel monkey

Immediate early gene expression in the cerebellar vermis of cats and squirrel monkeys was stimulated by prolonged whole body rotations. Continuous, earth-horizontal axis rotations that excited only otoliths or high velocity vertical axis rotations that excited only semicircular canals resulted in c-fos immunoreactive nuclei concentrated in the granular layer of lobules X and ventral IX (the nodulus and ventral uvula), which represent the medial parts of the vestibulo-cerebellum. Large clusters of labeled nuclei consisting mainly of granule cells and calretinin-positive unipolar brush cells were present in the granular layer, whereas Purkinje cell nuclei were unlabeled, and labeled basket and stellate cell nuclei were scattered in the molecular layer. In other vermal lobules there was a significant but less dense label than in the nodulus and ventral uvula. Generally, the extent of c-fos labeling of molecular layer interneurons was in relation to nuclear labeling of granular layer neurons: labeling of both basket and stellate cells accompanied nuclear labeling of neurons throughout the depth of the granular layer, whereas only stellate cells were labeled when nuclear labeling was restricted to the superficial granular layer. Yaw horizontal or roll vertical rotations each stimulated c-fos expression in the cat medial vestibulo-cerebellum to approximately the same extent. Low-velocity rotations resulted in much less c-fos expression. Similar, albeit less intense, patterns of c-fos activation were observed in monkeys. Concentrated c-fos expression in the medial vestibulo-cerebellum after exposure to a strong head velocity signal that could originate from either otolith or canal excitation suggests that granule and unipolar brush cells participate in a neuronal network for estimating head velocity, irrespective of the signal source.Gabriella Sekerková and Ema Ilijic contributed equally to the work     

Basket cells in the monkey fascia dentata: A Golgi/electron microscopic study

Summary This study describes non-granule cells in the fascia dentata of rhesus monkeys and baboons. Their cell bodies are located in the molecular layer and at the hilar border of the granular layer. They are called basket cells since their axons give rise to collaterals that branch in the close vicinity of the parent cell body and form symmetric synapses with dendrites and cell bodies of granule cells. These neurons are further classified with regard to the shape and location of their cell bodies and the orientation of their dendrites. Basket cells in the molecular layer are mainly bipolar with dendrites oriented perpendicular to the granular layer. These dendrites are densely innervated by presynaptic boutons forming asymmetric synapses. We have rarely observed molecular layer basket cells with dendrites traversing the granular layer and invading the hilus. We thus conclude that these cells are mainly activated by extrinsic afferents terminating in the molecular layer. Basket cells at the hilar border display pyramidal, fusiform or multipolar cell bodies that give rise to apical dendrites traversing the molecular layer and basal dendrites invading the hilar region. Large boutons establish asymmetric synapses with identified basal dendrites of these neurons. The dendrites of all types of basket cell are smooth, i.e. they had few or no spines. Many of them display varicosities. Cell counts in Cresyl Violet-stained sections revealed a ratio of basket cells to granule cells of 1:500. Essentially, the types of basket cell in the monkey fascia dentata are similar to those described previously for the rat. This contrasts sharply to our recent findings for pyramidal neurons and granule cells of the monkey hippocampus which showed an increased complexity and variability when compared with rodents. These data do not support the hypothesis that only local circuit neurons evolve in phylogeny.     

蜥蜴类卵泡的卵黄形成

目的：研究蜥蜴类卵黄形成的过程及机理。方法：用组织学、组织化学及免疫组织化学手段研究。结果：(1)无蹼壁虎的卵泡伴随着颗粒细胞发育及凋亡，卵黄由均质的构造转变为短丝状卵黄纤维，再变为由3种卵黄小板构成的卵黄；(2)北草蜥的卵泡，由具致密核心囊泡状卵黄变为由1种卵黄小板构成的卵黄；(3)无蹼壁虎卵泡鞘紧靠颗粒层的内层细胞碱性磷酸酶(ALP)呈强阳性反应，梨状细胞、中间型及小细胞ALP呈弱阳性反应；(4)P53在颗粒细胞内呈胞浆分泌型，甲胎蛋白(AFP)则主要分布于卵泡鞘。结论：ALP，AFP及P53蛋白在爬行类卵泡内表达部位及时间差异性与早期卵泡细胞与颗粒细胞分化和凋亡相关，颗粒细胞凋亡与解体可能启动了卵黄小板的形成。     

Substance P- and enkephalin-immunoreactive neurons in the hippocampus and related areas of the human infant brain

Neurons strongly immunoreactive for substance P are present as subpopulations in the stratum oriens, pyramidal layer and polymorphic layer of the hippocampus of the human infant. Substance P immunoreactive terminals are numerous on other neuronal cell bodies in the polymorphic layer and over pyramidal cells of the subiculum and the CA1, 2 and 3 regions. There is a high density of substance P-immunoreactive axons in the granular layer. Enkephalin immunoreactive neurons are relatively few in number and are present in the polymorphic and pyramidal layers. The results indicate that substance P probably plays a major role in short range circuits in the human hippocampus and that intrinsic enkephalin neurons probably play a relatively minor role.