中脑红核突触素的表达与突触发育的关系

目的:为探讨突触素在不同周龄阶段胎儿中脑红核的表达及其与红核突触发育的关系.方法:收集因故终止妊娠的胎儿32例,胎龄为16～39周,按周龄大小分为5组.利用免疫组织化学方法观察突触素在不同周龄阶段胎儿红核的表达情况,应用计算机图像分析技术测量突触素在不同周龄阶段胎儿红核表达的平均光密度.同时中脑取材,常规透射电镜技术处...     

胎儿端脑突触素表达和突触发育的研究

为了探讨突触素(SYN)在不同周龄阶段胎儿端脑额叶中的表达与胎儿额叶皮质突触发育的关系,本研究采用免疫组织化学方法观察突触素在不同周龄阶段胎儿额叶的表达水平,利用计算机图像分析技术测量不同周龄阶段胎儿额叶突触素表达的平均光密度;同时取材、常规电镜技术处理、透射电镜观察额叶突触发育的超微结构变化。结果显示:(1)光镜下各组均可见SYN免疫阳性产物主要表达于胎儿的额叶皮层,其表达量随周龄的增加而增强,各组间呈现显著性差异(P<0.05),其中16～24周胎儿额叶的阳性产物位于神经元的胞浆内,呈均匀的浅黄色,神经元突起内未见阳性产物;25～29周额叶的阳性产物呈黄色,在胞浆和突起内均可见,但阳性产物的量却下降;而30～39周额叶的SYN阳性产物呈棕黄色的点状或颗粒状,主要位于神经元的突起内,神经元胞浆内未见阳性产物,阳性产物的量显著增加;(2)透射电镜下19～36周胎儿大脑额叶均可见到突触样结构,随着周龄的增加,突触的数量逐渐增多,结构逐渐清晰和完整。上述结果提示SYN的表达可以反映胎儿神经系统发育的程度,SYN的表达与突触的发育是一致的;SYN在胎儿大脑额叶的表达部位经历由神经元胞浆内表达为主到神经元终末表达为主的这一过程,可能是由于SYN先是在神经元胞浆内合成,再随着神经元的发育而逐步转移到神经元突起的末梢部位。     

突触素在大鼠嗅球的分布

用免疫组织化学方法观察突触素在大鼠嗅球各层的分布。结果显示：小球层染色最深，其次为分子层，再其次是颗粒层，僧帽细胞胞体周围未见有明显阳性颗粒，纤维层和髓层不着色。     

大鼠内直肌亚核在动眼神经核的分布

目的：观察大鼠内直肌亚核神经元在动眼神经核的分布。方法：采用HRP逆行标记法,结果：大鼠支配眼球内直肌的运动神经元分布整个动眼神经核,在该核中央偏腹侧密集成群,结论：大鼠动眼神经核内直肌亚核有其特殊的分布形式。     

苯丙胺对大鼠海马突触素表达和突触结构的影响

目的探讨苯丙胺对大鼠海马突触素(SYN)蛋白质表达及突触结构的影响。方法将72只SD大鼠随机分为对照组、生理盐水组和苯丙胺组。用Western blotting检测大鼠海马结构内突触素表达的变化、用透射电镜观察大鼠海马CA3区突触的形态结构的变化。结果①苯丙胺组大鼠28d、42d的SYN相对表达量较对照组和生理盐水组相应时间明显降低,差异有统计学意义(P0.05);苯丙胺组42d的SYN相对表达量较14d明显减少,差异有统计学意义(P0.05)。②发现苯丙胺组大鼠较对照组的突触结构模糊、突触前后膜增厚,边界模糊,髓鞘出现松散分离样变。结论每天给大鼠肌注0.5mg/kg苯丙胺,28d出现海马结构突触素表达明显减少、海马CA3区突触结构明显损害;42d上述各项损害明显加重。     

小鼠晶状体发育过程中细胞增殖及突触素蛋白的表达

目的 探讨小鼠晶状体细胞增殖、细胞周期蛋白（cyclin) D1和突触素（SYN）的表达情况。方法 各日龄小鼠共150只,HE染色和
4’6-二脒基-2-苯基吲哚（DAPI）染色,在光镜下观察小鼠晶状体的一般结构;用抗细胞增殖核抗原（PCNA）免疫荧光法以及5′-溴脱氧尿嘧
啶核苷（BrdU）技术标记晶状体增殖细胞;用免疫荧标记标价法检测细胞周期蛋白cyclin D1和SYN在晶状体的表达。结果 孕8d（E8）左右,
晶状体由最初的晶状体板分化而来,是由单层柱状上皮细胞围成的空泡,空泡再进一步分化,被原始晶状体纤维所替代,逐渐形成晶状体;
BrdU和PCNA的阳性细胞主要分布在晶状体上皮细胞的前囊中央区,但出生10d（P10）以后BrdU阳性细胞已不在晶状体上皮细胞分布,而PCNA持
续表达。Cyclin D1阳性细胞在胚胎期主要在晶状体上皮细胞的赤道部表达,出生早期在晶状体上皮细胞的前囊中央区,出生5d（P5）以后在赤
道前区分布,赤道部有少量阳性细胞;在晶状体发育过程中,SYN可以在晶状体基质中和赤道部胞体中发现。结论 细胞增殖对晶状体的发育和
修复起着重要的作用;cyclin D1可能参与调控赤道部细胞的消失,因此保持晶状体的透明度;SYN在晶状体的表达说明了它对晶状体的透明有
着重要的作用,通过调控钙离子内流入胞体和基质中。     

LIF、CNTF和BDNF对动眼神经切断后猫动眼神经核CGRP表达的影响

为观察外源性白血病抑制因子(LIF)、睫状神经营养因子(CNTF)和脑源性神经营养因子(BDNF)对动眼神经(ONe)切断后猫动眼神经核(ONu)降钙素基因相关肽(CGRP)表达的影响,本研究采用猫ONe切断后硅胶管套接模型,向再生室内分别给予LIF、CNTF、BDNF、BDNF+LIF、BDNF+CNTF和生理盐水(NS),存活12周后用免疫组织化学方法检测各组动物ONu内CGRP的表达变化。结果发现:各营养因子组动物术侧ONu内的CGRP表达均明显高于NS组(P<0.01)。BDNF+LIF组与BD-NF+CNTF组之间无明显差别(P>0.05),但均明显高于因子单用组(P<0.01);因子单用组中CNTF组与BDNF组之间无明显差别(P>0.05),但均高于LIF组(P<0.05)。以上结果提示,外源性LIF、CNTF和BDNF均可上调ONe切断后猫ONu的CGRP表达,LIF作用较弱,联合应用具有协同作用。     

动眼神经压榨损伤后CGRP在SD大鼠动眼神经核中的表达

目的研究降钙素基因相关肽(calcitonin gene-related peptide,CGRP)在动眼神经损伤后再生过程中面神经运动神经元中的表达变化。方法健康SD大鼠分别行左侧动眼神经压榨术,术后饲养3、7、14、21、28、35d,取出脑干含动眼神经核团部分,用免疫组化及图像分析技术,观察舌下神经核中的CGRP在舌下神经再生中的变化。结果 CGRP分布于正常SD大鼠动眼神经各亚核,动眼神经损伤后3 d,损伤侧的动眼神经核中CGRP比对照侧增强。图像分析CGRP灰度值与对照侧比较,差异有统计学意义(P<0.05);损伤后7 d达最高峰(P<0.05),以后尽管显著表达但渐减。结论损伤导致CGRP在面神经运动神经元中的表达增加,提示CGRP在面神经再生修复过程中发挥调理作用。     

雷公藤内酯醇对阿尔茨海默病模型大鼠海马突触素表达及突触超微结构的影响

目的:探讨雷公藤内酯醇对阿尔茨海默病模型大鼠海马突触素表达及突触超微结构的影响.方法:大鼠随机分成对照组、模型组、治疗组.模型组给予双侧海马各一次性注射凝聚态Aβ1-4010μg,治疗组在海马注射凝聚态Aβ1-40后,每日腹腔注射雷公藤内酯醇0 4mg/kg, 15d后用免疫组织化学方法和蛋白免疫印迹技术检测海马突触素表达情况,透射电镜观察突触结构的变化.结果:与模型组相比,治疗组海马区突触素免疫反应阳性产物数量(152 80±15 76)及平均光密度(0 3180±0 0278)均增加;突触素表达总量(1917 71±41 02)及密度比值(0 87±0 03)亦增加;突触结构较清晰,界面增长,突触后电子致密物增厚.结论:雷公藤内酯醇可以增加阿尔茨海默病模型大鼠海马突触素的表达,减轻阿尔茨海默病模型大鼠海马突触损伤程度.     

突触素在大鼠嗅球的分布

用免疫组织化学方法观察突触素在大鼠嗅球各层的分布。结果显示:小球层染色最深,其次为分子层、再其次是颗粒层。僧帽细胞胞体周围未见有明显阳性颗粒,纤维层和髓层不着色。     

大鼠展神经核向动眼神经核的兴奋性和抑制性投射研究

目的:研究展神经核向动眼神经核内直肌亚核投射的神经递质通路.方法:利用荧光金(fluoro-gold,FG)逆行追踪结合免疫荧光组织化学双标技术观察大鼠展神经核内FG逆行标记细胞和谷氨酸(glutamate,Glu)、γ-氨基丁酸(γ-amino-butyric acid,GABA)、谷氨酸受体1型(glutamate...     

胎儿下颌下腺内瘦素和瘦素受体的表达及定位

目的：观察胎儿下颌下腺内瘦素和瘦素受体的表达、分布及发育规律。方法：HE染色法和免疫组织化学SABC法。结果：胎儿下颌下腺内纹状管和小叶问导管上皮细胞呈瘦素及瘦素受体免疫阳性反应,免疫反应产物分布于导管上皮细胞胞质内,细胞核星阴性反应。在胚胎发育的不同阶段,免疫反应强度亦不等。结论：瘦素和瘦素受体表达于胎儿下颌下腺内,可能参与调节胎儿下颌下腺和胃肠的发育及功能活动。     

Afferents to the lateral reticular nucleus from the oculomotor region

Summary By means of retrograde axonal transport of the wheat germ agglutinin — horseradish peroxidase complex, a projection from the Edinger-Westphal nucleus to the lateral reticular nucleus was demonstrated in the cat. Following small tracer ejections in the main part of the lateral reticular nucleus, a significant number of labelled neurons were found bilaterally throughout the Edinger-Westphal nucleus. Most of the labelled cells were located on the ipsilateral side. The projecting neurons are spindle-shaped to round with a maximum diameter of the cell body between 15 and 30 m. The findings are discussed in relation to other Edinger-Westphal efferent projections, and some comments are made concerning the cytoarchitecture and delineation of the feline Edinger-Westphal nucleus.     

Antidromic identification of midbrain near response cells projecting to the oculomotor nucleus

Summary Medial rectus motoneurons carry both conjugate and vergence eye position signals. Abducens internuclear neurons, whose axons travel in the medial longitudinal fasciculus, provide these motoneurons with the major signal for conjugate eye movements but not for vergence eye movements. A vergence signal appropriate for these motoneurons is seen on the near response cells that are found in the mesencephalic reticular formation within 2 mm of the oculomotor nucleus. The goal of the present study was to determine if midbrain near response cells project to the medial rectus subdivision of the oculomotor nucleus. Near response cells were recorded in two trained rhesus monkeys with ocular search coils. A stimulating electrode was positioned within the medial rectus subdivision of the oculomotor nucleus. Twenty-eight near response cells were found that could be driven by single pulse microstimulation of the ipsilateral medial rectus subdivision. In all cases, antidromic activation was confirmed by collision testing. Attempts to antidromically activate midbrain near response cells from the contralateral medial rectus subdivision were unsuccessful. Most antidromically activated cells had a steady state firing rate proportional to vergence angle. One cell also showed burst activity during the vergence eye movements. Divergence cells were not antidromically activated.     

人胎胃壁瘦素及瘦素受体的免疫组织化学

目的：对不同胎龄胎儿胃壁组织发育和胃壁内瘦素及瘦素受体进行定性和定位观察。方法：采用HE和免疫组织化学SABC染色方法。结果：胚胎13w,可见有胃小凹和胃腺。15w时,胃壁具有4层结构。随胎龄增长,胃壁和胃腺逐步发育完善。胚胎13w时,胎儿胃粘膜上皮和胃腺内壁细胞呈瘦素及瘦素受体免疫反应中等阳性,免疫反应产物分布于胞质,而胞核为阴性。随着胃腺的发育,呈免疫反应阳性的壁细胞数目增多。在胎儿整个发育过程中,各胎龄间胃腺瘦素及瘦素受体的免疫反应强度无明显变化。结论：人胚胎发育时期,在胃上皮和胃腺中有瘦素及瘦素受体的表达,推测它们对胎儿的生长发育起重要的调节作用。     

Development of the interpeduncular nucleus in the midbrain of rhesus monkey and human

Summary The development of the interpeduncular nucleus (IPN) in primates was studied in rhesus monkey with ³H-thymidine autoradiographic, Nissl and Golgi methods and in humans in histological preparations from embryos and fetuses of different ages. Autoradiographic analysis demonstrated that the neurons of the monkey IPN underwent their final cell division between postconception day 36 (E36) and E42, which corresponds to Stages 17 through 21 of Hendrickx and Sawyer. Autoradiograms of monkeys sacrificed at various short intervals following exposure to a pulse of ³H-thymidine showed that IPN neurons were generated in the proximity of the ventricular surface near the confluence of the 3rd ventricle and cerebral aqueduct, migrated ventrally along the midline and then spread laterally after reaching the ventral midbrain, where IPN was first recognized at E45 (Stage 23). The distribution of successively generated neurons in autoradiograms revealed caudal to rostral and lateral to medial spatio-temporal gradients. Differentiation of IPN neuronal size and development of Nissl substance began in rhesus monkey only after postmitotic cells had reached their destination and seemed to be pronounced mainly through E104. However, growth of the dendrites and elaboration of their side branches as seen in Golgi impregnations progressed gradually from E81 to birth (E165) and perhaps even later.Analysis of histological preparations of a series of human embryos and fetuses was used to derive similar information indirectly, since the autoradiographic method cannot be applied to man. It was found that IPN neurons in human probably underwent their final division between Carnegie Stage 17 and 21. Similarly, as in monkey, postmitotic cells in human IPN displayed an inverted fountain pattern of cellular migration. IPN could first be delineated at Stage 23. There was evidence for both caudal to rostral and lateral to medial spatiotemporal gradients in the human, as in the monkey. Thus, in monkey and human, all IPN neurons are generated within the first quarter of intrauterine life and there is remarkable similarity in the timing, tempo and pattern of IPN neuronal differentiation in both species, indicating the validity of using non-human primates as an experimental model for understanding the development of this structure in man.Supported in part by NIH grants NS12236, NS11233, 12265, HD08658, and RR00169.Mr. Halfon is a medical student at the University of California, Davis.     

胎儿肛管微血管分布特点及其临床意义

目的：研究胎儿肛管微血管的分布特点,为探讨肛门缺血性改变提供微血管的形态依据。方法：墨汁灌注,组织揭层透明铺片及组织切片,光镜下测量胎儿肛管肛门内括约肌及肛膜下区毛细血管网眼密度。结果：胎儿肛管在后中线处肛门内括约肌和粘膜下层存在毛细血管低密度区;肛门内括约肌里微动脉网的干动脉—肌束间动脉走行方向与肌纤维走行方向相垂直,括约肌强度收缩易压迫肌束间动脉,致血供不足。结论：肛管后中线处血供不良和肛门内括约肌痉挛也许是原发性肛裂好发于后连合处的主要原因。     

Radioautographic study of 3H-GABA uptake in the oculomotor nucleus of the cat

Summary The uptake of tritiated -aminobutyric acid (³H-GABA) in the oculomotor nucleus of the cat was studied, using light and electron microscopic examination of radioautograms after intracerebral in vivo administration of the amino-acid. A glial uptake by oligodendrocytes was seen together with a neuronal uptake of the tracer in a certain type of axon terminals found in synaptic contact with both dendrites and soma, some of them exhibiting all the ultrastructural features of motoneurons. Previous neurochemical, electrophysiological and immunocytochemical studies indicate that GABA might well be the inhibitory neurotransmitter in the vestibuloocular reflex arc. The present results show that a morphological substrate exists for the presumed postsynaptic GABAergic inhibition of ocular motoneurons, at least in the oculomotor nucleus of the cat.Supported by grant ATP 3650 from the Centre National de la Recherche Scientifique     

Afferents to the lateral reticular nucleus from the oculomotor region

Summary By means of retrograde transport of the wheat germ agglutinin-horseradish peroxidase complex, afferent fibres to the lateral reticular nucleus from the oculomotor and accessory oculomotor nuclei were demonstrated in the cat. Small iontophoretic ejections were made into the main part of the lateral reticular nucleus from a ventral approach. Significant numbers of retrogradely labelled neurons were found bilaterally in all parts of the oculomotor nucleus. The majority was of small size and distributed along the dorsal and lateral boundaries of the nucleus. Some labelled neurons were located just outside these boundaries, in the periaqueductal gray and the adjacent mesencephalic reticular formation. Retrogradely labelled neurons were also found in the accessory oculomotor nuclei: The interstitial nucleus of Cajal featured a substantial number of labelled neurons. Some labelled neurons were consistently found also in the nucleus of the posterior commissure, but no labelled neurons were found in the nucleus of Darkschewitch. The labelled neurons in the interstitial nucleus of Cajal were of different sizes and located bilaterally, mainly in its rostral part. Caudal as well as rostral parts of the main lateral reticular nucleus appear to receive the descending afferents from the oculomotor region, but higher numbers of labelled neurons were found subsequent to ejections in the rostral part. The findings are discussed and some comments are made concerning the lateral reticular nucleus as a possible relay nucleus for oculomotor input to the cerebellum.     

胎儿侧脑室室管膜发育的免疫组织化学和电镜研究

目的：探讨胎儿脑发育过程中出现的放射状胶质细胞、伸展细胞和普通室管膜细胞之间的关系。方法：运用HE染色、免疫组化染色和电镜观察胎儿侧脑室室管膜组织。结果：12w时可见放射状胶质细胞和位于室管膜层的神经干细胞样细胞。少数放射状胶质细胞有单纤毛;20w时室管膜胶质纤维酸性蛋白(GFAP)呈阳性,伸展细胞出现在室管膜层;28w大量GFAP阳性的星状胶质细胞位于室管膜下层和中间层;以后GFAP阳性强度和细胞数开始下降。39w时GFAP阴性,无法见到伸展细胞。结论：放射状胶质细胞主要转变为星状胶质细胞,少量转化为普通的室管膜细胞。伸展细胞可能来源于神经干细胞,部份转化为普通室管膜细胞;部分可能转化为其他细胞,可能是室管膜瘤中伸展细胞的来源。