Sugiyama Yuki, Prescott Alan R, Tholozan Frederique M D, Ohno Shigeo, Quinlan Roy A
School of Biological and Biomedical Sciences, Durham University, Science Park, Durham, UK.
Exp Eye Res. 2008 Jul;87(1):64-70. doi: 10.1016/j.exer.2008.03.017. Epub 2008 Apr 3.
The lens epithelium possesses an apical junctional complex (AJC) comprising adherens and tight junctions (AJs and TJs) and yet several key structural components and associated regulatory proteins have not been identified or localised in these cells. Here we determine the subcellular distribution of the archetypal TJ markers (ZO-1, claudin-1, and occludin) and TJ-associated cell polarity proteins (aPKC, Par3 and Par6beta) with AJ markers, E- and N-cadherin. As seen in other polarised epithelia, all these markers were located by confocal immunofluorescence microscopy to the apical ends of the lateral plasma membranes of bovine lens epithelial cells at sites of cell-cell interaction. Using immunoelectron microscopy, we show that ZO-1 concentrated at "kissing points" between neighbouring cells and these data, when taken in the context of our confocal immunofluorescence microscopy and blotting data, suggest the presence of TJs within the AJC. Likewise, immunogold labelling for E-cadherin identified AJs within these AJCs. We also report aPKC immunogold labelling localised to the AJC. These data show that the AJC of lens epithelial cells are a composite of TJs and AJs.
晶状体上皮细胞具有一个顶端连接复合体(AJC),该复合体由黏着连接和紧密连接(AJs和TJs)组成,然而,这些细胞中的几个关键结构成分和相关调节蛋白尚未被鉴定或定位。在这里,我们确定了典型的紧密连接标记物(ZO-1、闭合蛋白-1和闭合蛋白)以及与紧密连接相关的细胞极性蛋白(非典型蛋白激酶C、Par3和Par6β)与黏着连接标记物E-钙黏蛋白和N-钙黏蛋白的亚细胞分布。正如在其他极化上皮细胞中所观察到的那样,通过共聚焦免疫荧光显微镜观察,所有这些标记物都位于牛晶状体上皮细胞侧质膜顶端的细胞间相互作用部位。利用免疫电子显微镜,我们发现ZO-1集中在相邻细胞之间的“接触点”,当结合我们的共聚焦免疫荧光显微镜和印迹数据来看时,这些数据表明AJC内存在紧密连接。同样,E-钙黏蛋白的免疫金标记确定了这些AJC内的黏着连接。我们还报告了非典型蛋白激酶C的免疫金标记定位于AJC。这些数据表明,晶状体上皮细胞的AJC是紧密连接和黏着连接的复合体。