大鼠丘脑下部室管膜上皮细胞间加压素和催产素神经元的免疫组织化学观察

本文应用免疫组化PAP技术观察了位于大鼠丘脑下部第三脑室室管膜上皮细胞之间的加压素(VP)和催产素(OXT)神经元。在视前区第三脑室空周,见有较多的OXT神经元和较少的VP神经元紧贴室管膜分布,其突伸至室管膜下神经毡,有的穿行于室管膜上皮细胞之间并多与室管膜上皮细胞接触。在室旁核平面,比较远离脑室的VP、OXT神经元发出长突起行向第三脑室,有的穿过室管膜上皮伸至第三脑室腔(图3)。在上述二区域也发现个别OXT及VP神经元的胞体位于第三脑室侧壁及底壁的宝管膜上皮细胞之间。此外,在视上核前部及结节部,VP神经元的树突,大多伸向脑的腹侧面,有的树突似乎伸入了蛛网膜下腔。上述这两种肽能神经元与室管膜上皮的密切位置关系,可作为VP、OXT神经元具有感受脑脊液的变化或直接向脑脊液释放其分泌产物等设想的形态学基础。     

波形蛋白和胶质细胞纤维酸性蛋白在成年大鼠伸长细胞中的表达

目的：研究波形蛋白(VIM)和胶质细胞纤维酸性蛋白(GFAP)在成年大鼠伸长细胞(TAS)中的表达,观察TAS的特点并探讨胶质细胞的分类。方法：采用免疫组织化学方法,显示第三脑室腹侧壁和正中隆起(ME)TAS的特征。结果：在TAS中VIM的表达强度高,GFAP表达较弱,差异非常显著。对星形胶质细胞(AST)来说,不论使用Triton与否,GFAP均为高强度表达,而VIM为阴性。ME的TAS较第三脑室的突起较粗大分支较多。结论：无Triton存在下,VIM可以明确定位并区分第三脑室区域2种主要的胶质细胞TAS和AST;成年大鼠TAS还保持部分未成熟细胞的特性;ME是大量获得TAS的组织来源。     

大鼠脑室系统伸长细胞的顺行追踪研究

用霍乱毒素亚单位与辣根过氧化物酶结合物（ＣＢ－ＨＲＰ）对脑室系统伸长细胞进行顺行追踪。结果在第三脑室壁腹侧２／３区，大量的伸长细胞发出浓度的突起，对称伸入下天脑，甚至到达正中隆起胜利侧软膜表面，其中部分害起可直接与下丘脑内的血管相接触。在侧脑室壁、第四脑室底和脊髓中央管也发现有伸长细胞，但数量远较第三脑室壁少。本实验表明，伸长细胞不仅存在于第三脑室壁，而且在脑室系统的其他部位也有分布。这一结果提示     

后叶加压素在下丘脑的分布和释放途径及失水对正中隆起中后叶加压素的影响

我们用免疫细胞化学方法(ICC)对大白鼠下丘脑中加压素(VP)免疫反应阳性的神经元作了较详细研究。观察到VP阳性神经元存在于下丘脑室旁核(PVN)各亚核、视上核(SON)、交叉上核(SCN)、室周核(PN)及一些附属核团,包括环状核(CN)、交叉后核(RCN)、下丘脑外侧核(HLN)、穹窿周围核(PFN)。且发现一特殊的VP阳性胞体聚集区—很可能是多巴胺神经元聚集区A_(14)中的细胞,位于第三脑室侧壁中1/3段两侧,腹内侧核背侧。本文首次观察到在PVN和SON之间有VP阳性的神经纤维相联系。ICC和免疫电镜研究进一步证明在正中隆起外带存在VP阳性纤维,含大颗粒囊泡的VP末梢紧邻门脉毛细血管。将HRP注入第四脑室用HRP逆行追踪与ICC方法相结合,在PVN中观察到双标细胞,与直接用ICC法所见VP阳性轴突伸入第三脑室腔的结果一致。说明PVN中存在接触脑脊液神经元。干渴动物正中隆起的内、外带中的VP免疫反应明显减弱。     

大鼠下丘脑室旁核加压素神经元的生后发育

用免疫细胞化学PAP法研究了大鼠下丘脑室旁核加压素(VP)神经元的生后发育过程,对75只生后不同发育阶段大鼠下丘脑室旁核VP神经元进行了观察,结果表明,不同发育时期的室旁核内,均有VP免疫阳性细胞出现,细胞质和突起呈棕黄色,胞体大多数为椭圆形,少数为圆形和三角形,用测微尺测量了各级发育阶段阳性细胞的直径及计数了各切面被标记的细胞数量,大多数细胞只有二支突起,少数有三支突起。生后30天大鼠室旁槟VP阳性细胞的形态、数量、直径和复杂化程度已接近成年水平,另在第三脑室周圍灰质区域,室管膜和室管腆下层也发现有VP阳性细胞存在。     

自发性高血压大鼠第三脑室触液神经元内一氧化氮合酶与加压素的共表达

目的:观察自发性高血压大鼠(SHR)与SD大鼠第三脑室触液神经元(CSFCN)内一氧化氮合酶(NOS)与加压素(VP)的分布和共存。方法:应用还原型尼克酰胺嘌呤二核苷酸磷酸脱氢酶(NADPH-d)组织化学方法,结合ABC免疫组织化学技术。结果:在SHR视前区至室间核后大细胞亚核平面的第三脑室室壁均有NOS阳性CSFCN的分布;在SHR第三脑室室壁的CSFCN内NOS与VP具有共存性,且SHR组第三脑室的CSFCN内NOS与VP共存率较SD组高。结论:一氧化氮(NO)与VP在下丘脑的血压神经内分泌活动调节中起着重要的介导作用,也可能对高血压的发生发展有影响。     

伸长细胞对脑室注射的~3H—P物质吸收和转运的特异性的研究

本实验室已发现,在正常生理状态下脑室管膜某些区域的伸长细胞呈P物质(substance P,SP)免疫反应阳性。本实验采用同位素示踪放射自显影方法观察室管膜伸长细胞能否特异性吸收和转运脑室注射的~3H—SP,以进一步探讨这些SP免疫反应阳性伸长细胞中的SP是否为外源的。结果显示,注射~3H—SP后,脑室各段室管膜细胞(包括伸长细胞)内均有一定浓度的~3H—SP银粒分布。以秋水仙素阻断室管膜细胞轴浆运输,脑室各段室管膜上皮细胞表面和细胞内及室管膜上皮细胞下银粒数增多,而远离脑室的神经组织内银粒数相对较少。以上结果提示,伸长细胞(除延髓中央管)和普通室管膜细胞一样,均具有从脑脊液(cerebrospinal fluid,CSF)中主动摄取、转运~3H—SP的能力,但这种能力非伸长细胞所特有。由此推论,呈SP免疫反应阳性伸长细胞中的SP可能不是或不全是这些伸长细胞从CSF中吸收的。以上实验为深入研究伸长细胞的多种生理功能提供了实验依据。     

星型胶质细胞在帕金森病大鼠中脑、脑室下区的分布和体外培养分化特征

目的:观察星形胶质细胞在帕金森病(Parkinson's disease,PD)模型大鼠脑室下区、中脑的分布并进行PD模型大鼠脑室下区星形胶质细胞的体外培养、纯化和鉴定。方法:采用免疫荧光方法观察胶原纤维酸性蛋白(glial fibrillary acidic protein,GFAP)在PD模型大鼠中脑、脑室下区的表达;取PD模型大鼠侧脑室下区细胞进行体外培养、传代、贴壁分化和免疫荧光法鉴定。结果:(1)GFAP阳性细胞在PD模型大鼠中脑病侧较健侧明显增生,且GFAP荧光定量强度值在病侧较健侧明显增大(P0.05);GFAP阳性细胞在侧脑室、第三脑室和第四脑室室周区、视上核、下丘脑室旁核外侧大细胞部、正中隆起大量分布;(2)PD模型大鼠侧脑室下区培养分化的细胞具有星形胶质细胞的典型形态,其中原浆性星形胶质细胞突起粗短,分支多;纤维性星形胶质细胞突起细长,分支较少。结论:(1)星形胶质细胞在PD模型大鼠中脑和脑室下区的分布有明显区域性,可能与病侧的病理性增生相关,与相应区域的生理活动、调节功能及有关;(2)取PD模型大鼠侧脑室下区进行体外培养、分化获得星形胶质细胞的实验方法可靠。     

大鼠下丘脑第三脑室周催产素样神经元与脑脊液关系的电镜研究

本研究用高浓度戊二醛灌注固定、包埋前ABC免疫酶染色方法,显示了大鼠下丘脑第三脑室周与脑脊液密切相关的室管膜内及室管膜下OT-LI神经元丛。结果发现:在室旁核后大细胞亚核水平,第三脑室腔的顶及底部常有一些OT-LI轴突终末突入第三脑室腔,与脑脊液相接触,其纤维来源不明;透射电镜下见:这些OT-LI轴突终末附于室管膜细胞之上或夹行于微绒毛之间,常与第三脑室室管膜形成一种类似突触的连接,绝大多数OT-LI轴突终末内含有大量线粒体,其轴突包膜均呈节段性增厚。在视前区至室间孔水平的第三脑室底部,可见到OT-LI神经元胞体突入室管膜细胞间,似部分与脑脊液直接接触,透射电镜下见此类OT-LI神经细胞仅隔一层极薄的室管膜细胞浆与脑脊液相接触。在第三脑室侧壁室管膜下常有较密集的室管膜下神经元树突丛,透射电镜下常见到紧贴室管膜下走行的OT-LI树突,而且往往有轴突与之形成轴-树突触。上述结果提示,室管膜下的OT-LI神经元及树突可能既接受脑脊液中各种信息物质的调控又受到一些传入神经轴突终末的突触性传入控制而实施其分泌活动;突入脑室腔的OT-LI辅突可能是脑脊液中OT的产生途径之一。     

生长抑素在大鼠脑室管膜中定位分布的免疫组织化学和原位杂交…

采用免疫组织化学ＡＢＣ法和地高辛－碱性磷酸酶标记原位杂交组织化学方法观察生长抑素（ＳＳ）在大鼠脑室管膜中的定位分布。首次证实含ＳＳ的室管膜伸长细胞存在于下丘脑视前区第三脑室腹侧壁上，含ＳＳｍＲＮＡ的室管膜细胞在下丘脑视前区和视上区分布，为研究室管膜在脑－脑脊液神经体液回路中的作用，及ＳＳ在中枢神经系统中的生理功能提供了化学解剖学和分子生物学依据。     

Development and structure of the radial glia in the postnatal rat brain

Summary Postnatal development of radial glial cells was examined in albino rats. Until the 10th postnatal day radial glial cells were seen in the lateral ventricles, in the third ventricle (throughout its whole extent), in the aqueduct and in the fourth ventricle. The morphological appearance of radial glial cells was very variable in the different regions.After the 10th day radial glial cells (tanycytes) were seen only in the wall of the third ventricle. According to their appearance it was possible to undertake a morphological grouping. Considerable changes of the morphology of individual tanycytes could be observed in the median eminence and in the ventral hypothalamus between the 5th and 21st days.It was found that the peripheral processes of tanycytes ended near the nerve cells or on the cell body, on capillaries of the hypothalamic nuclei or on the pial surface. In a number of cases one tanycyte process contacted both the blood vessels of the hypothalamus and the pial surface.In view of their morphology the tanycytes can be assumed to transport material between different extracellular spaces, and/or to excrete material.The radial glial cells of the lateral ventricles can serve as guides for the postnatally formed microneurons and later can either transform to astroglia or degenerate.Part of this work was presented at the Annual Meeting of the Japanese Anatomical Society in Tokyo, 1979     

烫伤大鼠室旁核和触液神经元中胆囊收缩素的变化

目的 明确胆囊收缩素在烫伤应激中的确切作用。方法 在大鼠清醒状态下给予标准的２５％Ⅲ度皮肤烫伤,伤后２４ｈ杀动物,用免疫细胞化学ＡＢＣ法显示ＶＰＮ和ＣＳＦ－ＣＮｓ的ＣＣＫ阳性神经元,结果用显微图像分析技术处理。结果 （１）烫伤大鼠ＰＶＮ代表平面ＣＣＫ神经元的总面积增大,免疫染色无明显变化。（２）正中隆起（ＭＥ）的ＣＣＫ免疫染色阳性纤维的面积密度无显著性差异而免疫染色减弱。（３）烫伤大鼠第三脑室室管     

GLUT3阳性神经细胞在大鼠脑中的分布及其与脑内葡萄糖监控系统的关系

本文利用抗ＧＬＵＴ３ Ｃ－末端部分合成肽的抗血清对大鼠脑中ＧＬＵＴ３ 阳性神经细胞的分布进行了免疫组织化学研究。结果表明,除在大脑皮层的广泛区域和海马ＣＡ 区存在大量的ＧＬＵＴ３ 阳性神经元外,皮层下的某些核团和区域的神经元以及第三脑室腹侧部的ｔａｎｙｃｙｔｅ 也表达丰富的ＧＬＵＴ３ 蛋白。有趣的是,这些核团的大部分参与构成脑内葡萄糖监控系统的神经网络,有些阳性细胞（如下丘脑外侧区,黑质的神经元和ｔａｎｙｃｙｔｅ ）是这一网络中对血液和脑脊液中葡萄糖浓度发生反应的化学敏感细胞。这一新发现提示ＧＬＵＴ３ 可能与血糖浓度的神经调控有关。     

Production of antisera to acidic fibroblast growth factor and their application to immunohistochemical study in rat brain

Antisera against acidic fibroblast growth factor purified from bovine brain were produced in rabbits and used for immunohistochemical study of the rat brain. When examined in an immunospot assay using a nitrocellulose membrane, the best antibody was capable of detecting 80 fmol of acidic fibroblast growth factor but failed to react even with up to 5 pmol of basic fibroblast growth factor.

Using this antiserum, the immunohistochemical distribution of acidic fibroblast growth factor was examined in rat brain. Acidic fibroblast growth factor-like immunoreactivity was localized mainly in a subpopulation of ependymal cells and tanycytes, as well as in some glial cells. Positive ependymal cells were observed throughout the walls of ventricles, including the third ventricle and cerebral aqueduct. Immunoreactive processes of tanycytes were found extending from the ventral wall of the third ventricle to the brain parenchyma and surface. The most intense immunostaining was observed in circumventricular organs such as the organum vasculosum laminae terminalis and the subfornical organ. Particularly in the latter organ, there was an extremely dense plexus of immunoreactive fibers and processes around the wall of capillaries.

The present results suggest that the effects of acidic fibroblast growth factor on brain functions may be exerted through the circumventricular organs and/or ependymal cells.     

弓状核和正中隆起的超微结构特征及其机能意义

本文用醛-锇酸固定法对大白鼠弓状核和正中隆起的超微结构特征进行了研究。观察结果表明:在弓状核内有暗细胞和亮细胞两种神经元,它们可能兼具产生多巴胺和释放激素的功能。第三脑室室管膜的伸张细胞纵贯正中隆起的各层,抵达门脉毛细血管的周围间隙。含神经分泌物质的神经末梢可穿行于室管膜细胞之间,也可止于正中隆起毛细血管基膜周围,甚至直接紧贴在“开放型毛细血管”的内皮窗孔上。在弓状核神经细胞的胞体和树突上有轴体突触和轴树突触。在正中隆起的纤维层内有无颗粒囊泡型及颗粒囊泡型两种轴轴突触。在正中隆起的栅状层内,伸张细胞的基突与含有大颗粒囊泡的神经末梢之间也存在着突触联系。上述超微结构特征提示:1.丘脑下部的释放(或抑制)激素可能经由两条途径释放:直接由神经末梢释放入门脉毛细血管或先释放入第三脑室的脑脊液,由伸张细胞摄取,再由伸张细胞的基突转运至门脉毛细血管;2.释放(或抑制)激素的合成、贮存、运输和释放等各个环节都受到神经的调节和控制。     

Tanycytes of the third ventricle of the neonatal rat: A golgi study

The morphology and vascular relations of tanycytes in the walls and floor of the third ventricle (3rd V) of neonatal rats (4–10 days of age) were studied with a modification of the rapid Golgi method (Valverde, '70). Serial frontal sections through the infundibular recess revealed that tanycytes may be assigned to two distinct populations based on location and structure. Tanycytes in the ventral one third of the walls as well as the floor of the third ventricle comprise the first group. The cell body of these tanycytes was subependymal in location and possessed two main processes. A centrally directed short neck-process abutted upon the ventricular lumen and often exhibited one or more protrusions extending into the cerebrospinal fluid (CSF). Issuing from the opposite pole of the cell body, a second smooth, relatively unbranched tail process either projected directly to the ventral surface of the brain or terminated in contact with capillaries of the median eminence. A second group of tanycytes was located more dorsally in the walls of the third ventricle. The somatic apical surface of this group generally abutted directly upon the ventricular lumen and exhibited protrusions of diverse configuration projecting into the ventrical lumen. Tanycytes of the latter group were also characterized by a tortuous tail-process which arched laterally and ventrally to terminate either in the adjacent neuropil or at the surface of the brain. Unlike those of the ventral group, these processes exhibited numerous appendages, varicosities and side branches, many of which ended as expansions in direct contact with blood vessels. In addition, segments of beaded afferent fibers of fine caliber were often seen to contact processes of both groups of tanycytes. The observed relationships between CSF, tanycytes and blood vessels support recent hypotheses of tanycyte involvement in neuroendocrine regulation, CSF composition and neuronal metabolism.