【摘要】 目的对比Thy-1.1标记大鼠视网膜神经节细胞(RGCs)的方法,评估Brn-3a是否可作为一种可靠的内源性标记物,标记并计数大鼠RGCs。方法 20只SD大鼠,随机选取大鼠一眼为Thy-1.1组,另一眼作为Brn-3a组,常规H-E染色镜下观察RGCs的形态,运用免疫组织化学法检测Brn-3a与Thy-1.1的表达情况,每只眼球取1张切片,在每张待测视网膜切片上,以视网膜后极部中点为起点,至近角膜缘视网膜止点处,将视网膜分为3等份,即中央区、中间区及周边区,每个区域视网膜随机取5个高倍视野,分别计数Thy-1.1、Brn-3a阳性细胞均数。结果光镜下正常大鼠的视网膜从内向外可见3个细胞核层,依次为神经节细胞层(GCL)、内核层(INL)、外核层(ONL)。GCL的细胞呈单层排列,较为整齐,胞核清楚,数目相对较少,INL和ONL呈多层排列,且细胞排列紧密,数目相对较多。2种标记物染色都仅仅存在于视网膜的GCL,但在大鼠RGCs中分布呈明显差异:Brn-3a主要在胞核中着色,Thy-1.1主要在胞质中着色;运用此2种标记物标记计数RGCs,各个相同区域内Thy-1.1和Brn-3a阳性细胞计数比较,差别均无统计学意义(P>0.05)。Thy-1.1组和Brn-3a组中央区、中间区和周边区两两比较,差别均有统计学意义(P<0.01)。结论在大鼠视网膜中,Brn-3a可以作为一种比较可靠的内源性标记物,标记并计数大鼠RGCs。更多还原

【Abstract】 Objective To identify Brn-3ais a reliable endogenous marker of RGCs by contrast with Thy-1.1expression of the characteristics and differences count in rats.Methods 20health adult SD rats were divided into two groups,one eye was selected randomly for group of Thy-1.1,at the other as group of Brn-3ain the same rat.Morphological structure was investigated by haematotoxylin-eosine staining of retinal paraflin section respectively.Expression of Brn-3aand Thy-1.1was measured by immunohistochemical method.RGCs were counted by immunolabelled cells.Results There are ganglion cell layer(GCL),inner nuclear layer(INL)and outer nuclear layer(ONL)in retina of rat.GCL was monolayer,alignment rarefaction.Brn-3a+cells were only present in the GCL and showed a spatial distribution comparable to that of Thy-1.1+cells:the Brn-3asignal was located in the nuclei,and the Thy-1.1signal was major in the cytoplasm.There was no significant difference between the numbers of RCGs by labeling of these two markers in the same area.There was a significant difference among in the numbers of RCGs by labeling of these two markers in the different area.Conclusion Brn-3acan be used as a reliable marker to identify and quantify RGCs.更多还原