兔视网膜中P物质样免疫反应神经元的发育

本实验用免疫细胞化学ABC法,研究了成年、新生和生后兔视网膜中P物质(SP)样免疫反应神经元的定位和发育。结果表明,成年兔视网膜SP样免疫反应细胞胞体位于内核层和节细胞层,胞突分布在内网层的第1、3、5亚层,偶见于视神经纤维层。细胞密度以视纹最高,从视纹向背腹视网膜边缘区密度渐变小。在新生兔视网膜已有SP阳性胞体和胞突出现,胞体主要位于节细胞层,突起在内网层第5亚层,但未形成连续网层,在第1亚层很少,第3亚层未见SP阳性突起。SP阳性细胞密度从新生到生后第4天增加,生后第6天到第12天细胞密度渐下降。生后第12天SP阳性胞体主要位于内核层。生后第20天,SP阳性细胞的形态、密度与分布已接近成年水平。上述结果提示,在兔视网膜中SP样免疫反应胞体和突起在生前已出现,生后继续发育,到生后20天后其形态发育已接近成熟。     

碱性成纤维细胞生长因子对谷氨酸诱导的豚鼠视网膜内Caspase-3和Bcl-2表达的影响

目的:探讨过量谷氨酸毒性损伤后视网膜天冬氨酸蛋白酶-3(Caspase-3)和凋亡相关蛋白Bcl-2的表达及碱性成纤维细胞生长因子(bFGF)对兴奋毒性损伤的保护作用.方法: 豚鼠随机分为正常对照组、谷氨酸损伤组、bFGF治疗组.采用免疫组织化学方法和图像分析技术,对各组豚鼠视网膜内Caspase-3和Bcl-2的表达进行检测.结果: 对照组Caspase-3无明显表达,损伤组在节细胞层、内核层、内界膜和外界膜等处Caspase-3表达阳性面积百分率和密度明显上调,bFGF治疗组Caspase-3表达明显降低.对照组Bcl-2在节细胞层、神经纤维层及内核层内有弱阳性表达,损伤后变化不明显,bFGF治疗组Bcl-2的表达阳性面积百分率和密度明显上调,不仅在节细胞层、内核层和外界膜,在内界膜、外网状层、外核层也有明显表达.结论: bFGF能选择性减少视网膜兴奋毒性损伤后Caspase-3的表达,增加Bcl-2的表达,bFGF抗过量谷氨酸诱导的节细胞凋亡的机制之一可能是调节Mǘller细胞及节细胞内Caspase-3和Bcl-2的表达.     

脑红蛋白在绵羊视网膜中的分布

目的:探讨脑红蛋白在绵羊视网膜的分布特征。方法:利用免疫组织化学显色SP法,观察脑红蛋白在健康成年绵羊视网膜中的分布情况。结果:脑红蛋白在绵羊视网膜的视神经纤维层、内网状层、外网状层和光感受器内节段中有强阳性表达,在视网膜的内核层和节细胞层有弱阳性表达,在视网膜外核层、光感受器外节段和色素上皮层中未见有阳性表达,内界膜、外界膜和视神经中亦有脑红蛋白阳性表达。绵羊视网膜脑红蛋白阳性表达的细胞类型主要有节细胞、双极细胞和光感受器细胞,其中节细胞的阳性表达定位于细胞质,胞核中未见表达。结论;除外核层、光感受器外节段和色素上皮层外,脑红蛋白在绵羊视网膜其他各层中均有表达,提示脑红蛋白在维持视网膜中氧平衡状态时发挥重要作用。     

大鼠视网膜表达生长抑制mRNA细胞的发生

目的：研究视网膜中表达生长抑素mRNA神经元的发生和发育规律。方法：原位杂交组织化学。结果：胚胎第十三天（E13），视网膜内层就可以检测到生长抑素mRNA阳性细胞。从E14到E17阳性细胞增加迅速，E17达到高峰，此时视网膜节细胞层大部分为生长抑素mRNA阳性细胞，细胞排列紧密，从E18开始，阳性细胞逐渐减少，细胞排列开始稀疏。出生后当（PND0）阳性细胞数显著下降，从PND0到PND15，阳性细胞继续减少，阳性细胞主要位于节细胞层，PND20时，阳性细胞的数目及分布均与成年动物相似，部分已迁移至内核层的内表面。细胞开始或停止表达生长抑素mRNA均是从视网膜中央开始，然后向周围。结论：首次发现生长抑素基因在胚胎期的节细胞中一过性表达，提示生长抑素在视网膜节细胞的产生，分化以及视觉通路的形成与成熟有关。     

大鼠视网膜神经元NOS与Parvalbumin的共存研究

用NADPH脱氢酶组化及Parvalbumin免疫组化双标记技术观察了正常大鼠视网膜一氧化氮合酶(NOS)与Parvalbumin(PV)的分布,结果显示NOS阳性神经元主要位于内核层内缘带第二列,少数位于节细胞层,胞体圆形/卵圆形,直径8～12μm,细胞一侧发出突起伸向内网层1、3、5亚层,以第3亚层最为明显,PV免疫反应(PV—Ⅰ)神经元位于内核层最内缘第一列,少数位于第二列、中间部及节细胞层,胞体卵圆形,直径6～10μm,由胞体一端发出突起伸向内网层第1、5亚层.神经纤维层可见PV～Ⅰ纤维.内核层内缘第二列可见少数双标阳性细胞,在它们的PV免疫反应胞质内散布有NOS颗粒.实验结果表明 NOS阳性神经元与PV～Ⅰ神经元均为无长突细胞,分属不同的亚型,少效PV～Ⅰ神经元属节细胞,个别双标细胞可能为另一种亚型的无长突细胞,提示NOS与PV在视觉信息传递中可能存在某些联系.     

狗视网膜内一氧化氮合酶的表达

目的：探索狗视网膜内ＮＯＳ阳性细胞的分布和类型。方法：应用ＮＡＤＰＨ－黄递酶组化方法。结果：视网膜内含两种ＮＯＳ阳性细胞,一种胞体较大,圆形成三角形,着色深,直径为１２～１５μｍ,大闰地节细胞层,由胞体发出２～３支突起伸向内网层,另一种胞体上,圆形成卵圆形,着色浅,直径为６～８μｍ、胞突短,伸向内网层,胞体主要位于内核层近内网层处,少数位于内核层中部。两种ＮＯＳ阳性细胞分布和反应度均不同。结论：大     

人视网膜小胶质细胞形态及分布的研究

本文用白细胞共同抗原（CD45）和巨噬细胞特异性抗原（CD68）的单克隆抗体，对24，28，30，33，36周胎龄及正常成人的视网膜进行标记。结果表明幼稚的阿米巴型，成熟的分支型和介于两者之间的中间型（依突起数可分为单极，双极，多极形的小胶质细胞）小胶质细胞均表达CD45，除分支型外，其余的小胶质细胞亦同时表达CD68，小胶质细胞分布于视网膜的内核层，内网层，节细胞层。在其分布的层内，细胞分布均匀，相邻的细胞之间无接触，节细胞层的细胞数量比内核层，内网层多，但内核层，内网层的细胞比节细胞层的细胞幼稚，形态上表现为胞体较大，细胞无分支或分支较少，无次级突起，同时发现随胎龄的增大，细胞数量增多，突起增多并变长变细，36周胎时已分化为分支型小胶质细胞，亦观察到血管周小胶质细胞和血管旁小胶质细胞，在成体视网膜小胶质细胞以分支型为主，同时也存在一系列形态较幼稚的CD68阳性细胞。     

大鼠小脑皮质NPY能神经元的形态与分布

目的　研究大鼠小脑皮质内NPY免疫阳性神经元的形态与分布。方法　SD大鼠 10只 ,雌雄不限 ,升主动脉灌注固定 ,开颅取小脑 ,SP免疫组织化学染色。结果　大鼠小脑内NPY免疫阳性神经元分布在皮质的Purkinje细胞层 ,胞体呈梨形或烧瓶状 ,细胞核清晰不染 ,轴突伸向颗粒层 ,树突呈网状伸向分子层 ,突起未见染色。分子层和颗粒层未见阳性细胞 ,但可见神经纤维的存在。结论　小脑Purkinje细胞层内存在NYP阳性神经元。     

人胎视网膜细胞凋亡与小白蛋白免疫阳性神经元的发育

目的：观察人胎视网膜细胞凋亡与小白蛋白Parvalbumin,PV)免疫阳性神经元的分布与发育。方法：不同孕龄的人胎16例,TUNEL法标记凋亡细胞,ABC免疫细胞化学方法观察PV免疫阳性神经元的发育。结果：（1）细胞凋亡观察：12周人胎视网膜未见凋亡细胞;15周、17周凋亡细胞较多,大小不一,分布于视网膜的全局;20周凋亡细胞主要集中在内核层,数量减少;28周凋亡细胞仅见于内核层,呈指环样外观,着色较深,数量较20周减少明显。（2）PV免疫阳性神经元发育：12周人胎视网膜未见PV免疫阳性神经元;15周、17周视网膜节细胞层和内核层有弱阳性的PV免疫阳性神经元的分布与20周相似,但内核层的数量减少,呈整齐的带状排列,着色增强。结论：在胚胎28周视网膜神经元之间的突触联系已基本建立,而PV免疫阳性神经元发育和细胞凋亡在时间和数量变化上的一致性提示Ca^2 在视网膜的发育中起重要作用。     

兔胎视网膜内P物质免疫反应神经元的发生

本文用免疫细胞化学ＡＢＣ法，研究了新西兰白兔１８、２２、２５、２６、２８和３０ｄ胎龄视网膜内Ｐ物质免疫反应（ＳＰＩＲ）神经元的发生。在胎龄１８和２２ｄ兔视网膜未见ＳＰＩＲ细胞体和纤维。在胎龄２５ｄ视网膜的节细胞层最先出现ＳＰＩＲ神经元，胞体浅染呈卵圆形，突起不明显，在神经纤维层偶见串珠状ＳＰＩＲ纤维，其平均细胞密度为１０４．６个细胞／ｍｍ￣２。到胎龄２６和２８ｄ时，在节细胞层的ＳＰＩＲ神经元的胞体渐深染，可见个别ＳＰＩＲ神经元发出粗而短的突起伸向内网层，平均细胞密度分别为３８７和７７９．５个细胞／ｍｍ２。到胎龄３０ｄ时ＳＰＩＲ神经元开始出现于内核层的内排细胞，但数量很少，胞体呈卵圆形，发出细突起伸入内同层，在节细胞层的ＳＰＩＲ神经元的突起分支增加。此时ＳＰＩＲ神经元平均细胞密度为３５７．４个细胞／ｍｍ￣２。     

锌转运体ZNT7在小鼠视网膜的定位分布

目的研究锌转运体-7(zinc transporter 7,ZNT7)在小鼠视网膜的定位和分布。方法应用免疫组织化学技术检测CD-1小鼠视网膜内的ZNT7免疫反应产物的表达。结果ZNT7在小鼠视网膜内分布广泛,在神经节细胞和色素上皮细胞内ZNT7免疫阳性反应产物的表达最丰富,在无长突细胞和视神经纤维层中ZNT7免疫阳性反应产物为中等程度的表达,在内网层、外网层和光感受器外节中ZNT7免疫阳性染色较淡,在外核层和光感受器内节中ZNT7几乎没有表达。结论ZNT7可能在维持视网膜锌稳态过程中起到重要的作用。     

Neuronal and glial localization of homocysteate-like immunoreactivity in the rat retina

Summary To study the distribution ofl-homocysteate in the rat retina, specific polyclonal and monoclonal anti-homocysteate antibodies have been used in combination with a highly sensitive postembedding method for light microscopic immunocytochemistry. In central and peripheral retina, the most strongly immunoreactive cell bodies lay in the inner nuclear layer. They represented about 17% of the total neuronal cell population of the layer and were identified as bipolar cells (19–20% of cells in the outer half of the inner nuclear layer) and amacrine cells (15% of cells in the inner half of the inner nuclear layer). A third cell type showing heavy homocysteate-like immunoreactivity was identified as Müller glial cells. Characteristically, their descending processes formed three immunoreactive bands in the inner plexiform layer. Furthermore, the outer and inner limiting membranes as well as glia around and between ganglion cell axons and in the vicinity of blood vessels were labelled intensely. Photoreceptors and their terminals, and ganglion cells, were not immunostained. These findings indicate the presence of homocysteate in some bipolar and amacrine cells of the inner nuclear layer and support a role for this sulphur-containing excitatory amino acid as a neurotransmitter candidate in the retina.     

Somatostatin-like immunoreactivity in amacrine cells of the chicken retina

Somatostatin-like immunoreactivity was detected in chicken retina by radioimmunoassay. The levels of somatostatin-like immunoreactivity decreased after intra-ocular injection of kainic acid, but were not affected by destruction of the ganglion cells. By immunohistochemistry, somatostatinimmunoreactive amacrine cells were found in the inner nuclear layer. These cells were destroyed by kainic acid. At least some of the cells projected to all three sub-layers of the inner plexiform layer in which there were diffuse bands of fluorescence. Specific immunofluorescence was also detected at the level of the outer limiting membrane and the optic nerve fibre layer, but the outer nuclear and plexiform layers, horizontal, bipolar and ganglion cells did not show specific immunofluorescence.It is suggested that other amacrine cell sub-classes, defined in terms of their putative transmitter, may show specific patterns of cell body location and size, and terminal arborisation.     

Synapsin I expression in the rat retina during postnatal development

Summary The expression of the synapsin I gene was studied during postnatal development of the rat retina at the mRNA and protein levels. In situ hybridization histochemistry showed that synapsin I mRNA was expressed already in nerve cells in the ganglion cell layer of the neonatal retina, while it appeared in neurons of the inner nuclear layer from postnatal day 4 onward. Maximal expression of synapsin I mRNA was observed at P12 in ganglion cells and in neurons of the inner nuclear layer followed by moderate expression in the adult. At the protein level a shift of synapsin I appearance was observed from cytoplasmic to terminal localization during retinal development by immunohistochemistry. In early stages (P4 and P8), synapsin I was seen in neurons of the ganglion cell layer and in neurons of the developing inner nuclear layer as well as in the developing inner plexiform layer. In the developing outer plexiform layer synapsin I was localized only in horizontal cells and in their processes. Its early appearance at P4 indicated the early maturation of this cell type. A shift and strong increase of labelling to the plexiform layers at P12 indicated the localization of synapsin I in synaptic terminals. The inner plexiform layer exhibited a characteristic stratified pattern. Photoreceptor cells never exhibited synapsin I mRNA or synapsin I protein throughout development.Abbreviations GCL ganglion cell layer - INB inner neuroblast layer - INL inner nuclear layer - IPL inner plexiform layer - ONB outer neuroblast layer - ONL outer nuclear layer - OPL outer plexiform layer     

Expression profile of heat shock protein 108 during retinal development in the chick

In the developing chick retina, heat shock protein 108 (HSP108), which exhibits transferrin binding activity, has been demonstrated at the mRNA level, while transferrin shows two expression peaks. Here, we investigated the expression profile of HSP108 in the developing chick retina at the protein level. The localization of HSP108 in embryonic days 15 (E15), E18, and postnatal day 2 (P2) chick retina was examined immunohistochemically using monoclonal antibody 9G10 specific for chick HSP108, while the expression levels of HSP108 in developing chick retina from E12 to P2 and adult were measured by Western blot analysis. HSP108 was expressed in the ganglion cell layer, inner nuclear layer, outer plexiform layer, outer nuclear layer, inner segments of photoreceptors and retinal pigment epithelium. Two peaks of HSP108 expression were found at around E13 and E18, respectively. Since the two HSP108 peaks appeared to be correlated with the transferrin expression peaks during retinal development, HSP108 may be associated with iron metabolism during the development of the retina.     

Histology of the ferret retina

The retina of the adult ferret, Mustelo furo, was studied with light and transmission electron microscopy to provide an anatomical basis for use of the ferret as a model for retinal research. The pigment epithelium is a simple cuboidal layer of cells characterized by a zone of basal folds, apical microvilli, and pigment granules at various stages of maturation. The distinction between rod and cone photoreceptor cells is based on their location, morphology, heterochromatin pattern and the electron density of their inner segments. The round, light-staining cone cell nuclei occupy the layer of perikarya along the apical border of the outer nuclear layer. The remainder of the outer nuclear layer consists of oblong, deeply-stained rod cell nuclei. Ribbon type synaptic complexes involving photoreceptor cell axons, horizontal cell processes, and bipolar cell dendrites characterize the outer plexiform layer. The inner nuclear layer is comprised of horizontal, bipolar, and amacrine cell perikarya as well as the perikarya of the Müller cells. The light-staining horizontal cell nuclei are prominent along the apical border of the inner nuclear layer. The light-staining amacrine cell nuclei form a more or less continuous layer along the basal border of the inner nuclear layer. Both conventional and ribbon-type synapses characterize the inner plexiform layer. The ganglion cells form a single cell layer. The optic fiber layer contains bundles of axons surrounded by Müller cell processes. Small blood vessels and capillaries are present in the basal portion of the retina throughout the region extending from the internal limiting membrane to the outer plexiform layer. The adult one-year-old retina is compared with the retina at the time of eye opening.     

高血压大鼠视网膜溶酶体酶组织化学研究

目的：探讨溶酶体酶在高血压视网膜网变发生过程中的作用。方法：应用光镜定量酶组织化学方法对ＷＫＹ大鼠和自发性高血压大鼠视网膜原酸性磷酸的分布和活性变化进行定量观察。结果：视网膜各层酸性磷酸酶活性岂强到弱依次是（Ｆ检验，Ｐ〈０．０５）；（１）色素上皮层；（２）视杆维层内节和外网层（两层间活性无显著性差异）；（３）内网层；（４）节细胞层和神经纤维层，（５）外核层和内核层（两层间活性无显著性差异）杆锥层外     

人胎视网膜发生的光镜和电镜观察

本文在光镜下观察40例人胎视网膜的发生,在电镜下观察15例人胎视网膜视细胞、双极细胞、节细胞的发育。结果表明:胚胎第9周时神经上皮可分内、外成神经细胞层。第10周时内、外成神经细胞层之间的Chievitz带消失;第11周时节细胞从内成神经细胞层内迁;第13周节细胞与内成神经细胞之间出现内网层;第16周始双极细胞从外成神经细胞层中内迁形成外网层和内核层。第20周后视网膜各层形成。而视细胞、双极细胞、节细胞的超微结构于胎儿8个月后才发育完善。其结构与成人基本相同。     

高血压大鼠视网膜组织Ca^2＋酸性磷酸酶组织化学的研究

目的：应用光镜定量酶组织化学方法对正常京都种大鼠（ＷＫＹ）和自发性高血压大鼠（ＳＨＲ）视网膜组织的Ｃａ＾２＋－酸性磷酸酶的分布和活性进行定量观察，结果：Ｃａ＾２＋酸性磷酸酶在ＷＫＹ视网膜组织的活性由强到弱依次为（Ｆ检验，Ｐ〈０．０５）；（１）杆锥细胞内节和外核层；（２）节细胞层；（３）内核层；（４）内网层和外网层；（５）杆锥细胞外节阴性，在ＳＨＲ视网膜组织中，各层Ｃａｓ＾２＋酸性磷酸酶活性下降，以