双侧眼球摘除对大鼠视交叉上核的影响—免疫组化及光电镜体视…

采用双侧眼球摘除大鼠模型，术后存活不同时间，用免疫组化法探讨视交叉上核内ＶＩＰ和ＡＶＰ免疫反应强度和面积的变化；对变化时明显的２１ｄ组进一步同光镜和电镜体视学方法进行形态学研究。＜１＞免疫组化研究：ＶＩＰ免疫染色，２１ｄ组免疫反应强度和面积均比对照组降低（Ｐ＜０．０５），２ｄ和７ｄ组免疫反应强度和面积与对照组相比无显著差异。ＡＶＰ免疫染色，三个时间实验组免疫反应强度和面积均与对照组无显著差异。＜２     

摘除松果腺对大鼠视交叉上核节律性的影响

成年Ｗｉｓｔａｒ雄性大鼠２０只，实验组１０只，行松果腺摘除术；对照组１０只。术后３０ｄ将实验组、对照组动物各半数在０９：００～１０：００和１６：００～１７：００分别处死。用免疫组化ＡＢＣ法显示视交叉上核内含ＶＩＰ的神经元；微机图像分析仪上测量其光镜下的切面面积及平均免疫反应强度。结果：（１）对照组不同时间处死的动物ＶＩＰ样神经元切面面积０９：００～１０：００大于１６：００～１７：００，呈节律性变化。（２）摘除松果腺后各时间组间ＶＩＰ样神经元切面面积的差异无显著性，提示实验组动物视交叉上核的ＶＩＰ样神经元功能活动的日周节律已发生改变。（３）对照组和实验组动物的ＶＩＰ样神经元平均免疫反应强度在所测的两个时间中均未见明显差异。     

摘除双侧眼球对大鼠视交叉上核节律性的影响

成年Ｗｉｓｔａｒ雌鼠５７只，随机分为实验组３０只，行双眼摘除术。对照组２７只。术后４周将各组动物体重相近的每３只列为一个配伍组，分别在０９：００～１０：００、１７：００～１８：００、２３：００～２４：００三个时间处死。将含有视交叉上核的脑组织经固定、恒冷箱切片后，用免疫组化ＡＢＣ法染色显示视交叉上核内含ＶＩＰ或ＡＶＰ的神经元，微机图像分析仪上测量光镜下这两种神经元的相对切面面积及平均免疫反应强度。结果：（１）对照组不同时间处死的动物ＶＩＰ能神经元切面面积以２３：００～２４：００最大，０９：００～１０：００次之，１７：００～１８：００最小，呈昼夜节律变化，ＡＶＰ能神经元也以２３：００～２４：００最大，但在０９：００～１０：００和１７：００～１８：００间差异无显著性；（２）摘除双眼后各时间组间ＶＩＰ能神经元和ＡＶＰ能神经元切面面积均不再显示显著性差异。提示实验组动物视交叉上核的这两种神经元功能活动的昼夜节律已发生改变；（３）实验组和对照组动物的ＶＩＰ能神经元和ＡＶＰ能神经元平均免疫反应强度在所测的三个时间中，均未见明显差异。     

大鼠视交叉上核内VIP、AVP及SOM样神经元的相互关系──免疫组化双重反应法研究

视交叉上核具有生物钟功能已为很多实验研究所证实，但其功能机制尚在探索之中。本实验采用双重免疫组织化学反应技术对大鼠视交叉上核内ＶＩＰ、ＡＶＰ及ＳＯＭ样三大神经元群之间的相互联系进行了观察．结果表明：（１）ＶＩＰ样扣结广泛分布于ＡＶＰ样神经元周围，数量最多、密度最大；而ＳＯＭ样扣结贴附于ＶＩＰ及ＡＶＰ样神经元的数量次之；（２）ＡＶＰ样扣结与ＶＩＰ样神经元之间，ＶＩＰ样扣结与ＳＯＭ样神经元之间也形成联系．上述发现为视交叉上核功能机制的研究提供了进一步的形态学依据．     

大鼠视交叉上核内VIP,AVP及SOM样神经元的相互关系——免疫组化…

视交叉上核具有生物钟功能已为很多实验研究所证实，但其功能机制尚在探索之中，本实验采用双重免疫组织化学反应技术对大鼠视交叉上核内ＶＩＰ，ＡＶＰ及ＳＯＭ样三大神经元群之间的相互联系进行了观察。结果表明：（１）ＶＩＰ样扣结广泛分布于ＡＶＰ样神经元周围，数量最多，密度最大，而ＳＯＭ样扣结贴附于ＶＩＰ及ＡＶＰ样神经元的数量次之；（２）ＡＶＰ样扣结与ＶＩＰ样神经元之间，ＶＩＰ样扣结与ＳＯＭ样神经元之间也形成联     

摘除侧眼球对大鼠视交叉上核节律性的影响

成年Ｗｉｓｔａｒ雌鼠５７只，随机分为实验组３０只，行双眼摘除术。对照组２７只。术后４周将各组动物体重相近的每３只列为一个配伍组，分别在０９：００－１０：００、１７：００－１８：００、２３：００－２４：００三个时间处死。将含有视交叉上核的脑组织经固定、恒冷箱切片后，用免疫组化ＡＢＣ法染色显示视交叉上核内含ＶＩＰ或ＡＶＰ的神经元，微机图像分析上测量光镜下这两种神经元的相对切面面积及平均免疫反应强度。结     

大鼠视交叉上核内VIP、AVP及SOM样神经元的分布及相互联系—免疫组化多重标记法研究

大鼠视交叉上核内ＶＩＰ、ＡＶＰ及ＳＯＭ样神经元的分布及相互联系──免疫组化多重标记法研究王蕾，欧可群，操高原，陈文玉华西医科大学成都６１００４１视交叉上核（ＳＣＮ）具有生物钟功能已为众多实验所证实，但对其节律发生并保持与外界同步的机制却尚在探索之中。...     

短期饥饿对老年,成年大鼠下丘脑加压素神经元及血浆皮质酮的影响

免疫组化光，电镜法观察禁食五天老年及成年大鼠下丘脑精氨酸加压素（ＡＶＰ）神经元，并检测其血浆皮质酮（Ｂ），与同龄正常鼠相比。结果：（１）Ｂ均值明显升高（Ｐ＜０．０１），（２）各神经核团的阳性神经元有增有减，非同步变化，（３）正中隆起外带阳性纤维密集，面积增加，（４）视上核主要及室旁核的神经元粗面内质网层次增多，线粒体较密集，（５）成年饥饿鼠阳性细胞总数增加，但其视上核主部的减少，并和血浆Ｂ的增加负     

大鼠中缝背核及中缝正中核内的VIP、GABA样触液神经元

本文将ＣＢ－ＨＲＰ注入侧脑室，用ＣＢ－ＨＲＰ逆行迫踪与免疫细胞化学相结合的方法，对大鼠脑干内的中缝背核及中缝正中核的远位触液神经元进行了定性研究。结果表明：中缝背核内存在ＶＩＰ样、ＧＡＢＡ样免疫反应阳性的触液神经元；中缝正中核内亦存在少量ＶＩＰ样、ＧＡＢＡ样免疫反应阳性触液神经元。它们的形态和数量各异。本文首次报道中缝背核和中缝正中核内远位触液神经元的化学性质，为探索其机能意义提供了形态学依据。     

大鼠视交叉上核与视上核的纤维联系—病毒跨神经元追踪,免疫组化双重标记法研究

大鼠视交叉上核与视上核的纤维联系—病毒跨神经元追踪，免疫组化双重标记法研究王蕾欧可群陈文玉操高原张军马玉琼（华西医科大学组织学研究室）有文献报道视交叉上核（ＳＣＮ）的传出纤维到达室旁核（ＰＶＮ），但是否有传出纤维投射至视上核（ＳＯＮ），目前尚未见报道...     

Contrary effect of eye enucleation on VIP-immunoreactive neurons in the suprachiasmatic nucleus and the superior colliculus of the rat

The topographic alternation of vasoactive intestinal peptide (VIP) immunoreactivity after long-term bilateral eye enucleation (80 days) was investigated in the suprachiasmatic nucleus (SCN) and the superior colliculus of the adult rat with the peroxidase-antiperoxidase method. In the SCN, the immunostaining of VIP-immunoreactive (IR) cell bodies increased, and the intensity and density of VIP-IR fibers and terminals were markedly enhanced after eye enucleation. On the other hand, after eye enucleation, no VIP-IR cell bodies and fibers were visible in the stratum griseum superficiale of the superior colliculus, although fusiform-shaped VIP-IR neurons were distributed in the layer of unoperated control rats. The present results, therefore, have revealed that the long-term blockage of retinal input induces changes of VIP immunoreactivity in different manners, according to the target brain areas.     

VIP neurons as prime synaptic targets for serotonin afferents in rat suprachiasmatic nucleus: a combined radioautographic and immunocytochemical study

Summary Cellular relationships between serotonin (5-HT) axon terminals and neurons containing vasoactive intestinal peptide (VIP) were characterized by combined radioautography and immunocytochemistry in rat suprachiasmatic nucleus (SCN). Light microscopic immunoradioautographs showed significant overlap between (³H)5-HT uptake sites and VIP-immunoreactive elements in the ventral half of the SCN. Of the 255 (³H)5-HT-labelled axonal profiles detected in a systematic electron microscopic survey of single thin sections from this area, 75 (30%) were directly apposed to VIP-immunoreactive nerve cell bodies and/or dendrites. Radioautographically labelled 5-HT varicosities often showed well-differentiated, symmetrical or asymmetrical synaptic junctions, 60% of which were established on VIP-immunoreactive nerve cell bodies or dendrites. In a separate sampling of 198 (³H)5-HT-labelled terminals seen in apposition with VIP-immunoreactive elements, 50 showed a junctional complex at the site of contact. Postsynaptic immunoreactive elements were mostly dendrites but also included nerve cell bodies. Despite the methodological limitations inherent to the present double labelling approach, these data strongly support the view that VIP neurons are prime synaptic targets for 5-HT afferents in the SCN. VIP/5-HT interactions are thus likely to play an important functional role in this nucleus and may in particular subserve the 5-HT mediated regulation of certain circadian rhythms, including that of pituitary hormone secretion.     

The Effects of Serotonin on the Differentiation of Neurons Producing Vasoactive Intestinal Polypeptide in the Suprachiasmatic Nucleus of the Rat

The morphogenetic influences of serotonin on the differentiation of neurons synthesizing vasoactive intestinal polypeptide (VIP) in the suprachiasmatic nucleus were studied in rats. This was addressed by comparative morphofunctional analysis of VIP neurons in adult rats whose brains developed prenatally in conditions of normal and deficient serotonin metabolism. Serotonin deficiency was created in fetuses by treatment of their mothers with p-chlorophenylalanine (PCPA). Pregnant females in controls were treated with 0.9% NaCl. VIP neurons in experimental and control animals were found to show no differences in VIP mRNA concentrations and, probably, in the level of VIP synthesis. However, inhibition of serotonin synthesis led to an increase in the number of VIP-immunoreactive neurons and an increase in the VIP concentration within these cells. This was not associated with any change in neuron size, which was an indicator of the absence of functional hypertrophy accompanying activation of specific synthesis. Comparison of the data obtained here showed that during prenatal ontogenesis, serotonin has an imprinting influence on the differentiation of VIP neurons and is probably involved in the formation of the mechanism of VIP secretion.     

Stereological quantitative analysis of LM and EM images on membranous glomerulonephritis (MGN) in rabbits

MGN model in rabbits was established according to Border's method. The pathological changes of MGN were analysed through stereological quantitation technique under LM and EM. Results indicate that the quantitative parameters of subepithelial electron dense deposits and the glomeruli in the experimental group were remarkably different statistically from those of the control group (P less than 0.05 or P less than 0.001). In the experimental group, the volume density of glomerular crescent or circumferential corpuscle was 17.39% at the 8th week of the experiment. The thickness of glomerular basement membrane was 168 nm in the control group and 627 nm in the experimental group (P less than 0.001).     

GABA neurons in the rat suprachiasmatic nucleus: Involvement in chemospecific synaptic circuitry and evidence for GAD-peptide colocalization

Summary Dual labelling methods were employed for the electron microscopic detection of glutamate decarboxylase (GAD) immunoreactivity, together with vasoactive intestinal peptide (VIP) or neuropeptide Y (NPY) immunoreactivity in the suprachiasmatic nucleus (SCN) of colchicine pretreated and untreated rats. These methods involved the combined use of diaminobenzidine and benzidine dihydrochloride as distinct chromogens to visualize peroxidase-anti-peroxidase (PAP) immunostaining, and a combination of the PAP procedure with a radioimmunocytochemical method employing¹²⁵I-labelled secondary antisera. We were thereby able to demonstrate that -aminobutyric acid (GABA) terminals provide an important afferent synaptic input to VIP neurons. Some of these VIP-immunoreactive neurons also exhibited GAD immunoreactivity. Examples of direct appositions between GABA and NPY terminals, and of a convergence of the two types of terminals on to the same postsynaptic targets, were frequently encountered. NPY/GAD colocalization within a few axonal varicosities was also demonstrated. These data provide additional information concerning chemospecific neuronal interactions that could be of functional importance in the regulation of circadian rhythmicity at the level of the SCN.     

大鼠垂体远侧部ACTH细胞与GH细胞形态学关系的免疫细胞化学研究

用光镜和电镜免疫细胞化学方法，研究了大鼠垂体远侧部ＡＣＴＨ细胞与ＧＨ细胞的形态学关系，以及在实验性胃溃疡状态下的变化。正常情况下，ＡＣＴＨ细胞与ＧＨ细胞彼此相邻，紧密接触，ＡＣＴＨ细胞的胞质突起可伸到ＧＨ细胞之间或包绕ＧＨ细胞，电镜下，有的ＡＣＴＨ细胞与ＧＨ细胞之间有可见有基板相隔，但更常见两者直接贴邻，没有基板分隔，并可见有些细胞之间有细胞连接。在实验性胃溃疡状态下，与ＧＨ细胞相邻的ＡＣＴＨ细胞     

Vasoactive intestinal peptide (VIP)-like immunoreactive neurons located in the rat suprachiasmatic nucleus receive a direct retinal projection

The existence of a direct projection from retinal ganglion cells to vasoactive intestinal peptide (VIP)-like immunoreactive neuronal elements in the rat suprachiasmatic nucleus (SCN) was revealed by combining analysis of degenerating axons following enucleation and electron microscopic immunocytochemistry. Degenerating axons appeared to make synaptic contact with VIP-like immunoreactive dendrite and neuronal perikarya in the ventral part of the SCN. The possibility of neuronal input from retinal ganglion cells to axons of VIP-like immunoreactive neurons was also suspected since axo-axonic synapses were detected between degenerating axons and axons with VIP-like immunoreactivity. Thus, VIP-like immunoreactive neurons in the SCN receive several neuronal inputs, including those from the retina, and may play a significant role in circadian entrainment.     

海马中56 kD蛋白诱导人神经干细胞向神经元分化的作用

在已证明切割海马伞海马中差异表达的56kD蛋白具有诱导神经干细胞迁移作用的基础上,进一步探讨其诱导神经干细胞向神经元和AChE阳性神经元分化的作用。取切割SD大鼠海马伞后14d及正常海马组织的匀浆进行非变性聚丙烯酰胺凝胶电泳(Native-PAGE)。将用无血清培养技术获取的人胚神经干细胞球分别与切割海马伞后14d和正常海马56kD差异蛋白的胶条及空白对照胶条共培养,在倒置显微镜下观察神经干细胞球中细胞的迁移和分化情况,于第21d分别应用MAP-2免疫荧光和AChE组织化学方法检测人神经干细胞分化为神经元和AChE阳性神经元的情况,并进行神经元的计数、细胞面积、细胞周长的图像处理和统计学分析。结果显示,分化的MAP-2阳性神经元的数量、细胞面积及细胞周长三项指标切割组最好,正常组次之,而空白对照组最差;三组AChE组织化学染色均为阴性结果。上述结果提示切割海马伞后海马中差异表达的56kD蛋白具有诱导神经干细胞向MAP-2阳性神经元分化的作用,但不能诱导神经干细胞向AChE阳性神经元分化。