Ultrastructural morphology and expression of proteoglycans, βig-h3, tenascin-C, fibrillin-1, and fibronectin in bullous keratopathy
Ultrastructural morphology and expression of proteoglycans, βig-h3, tenascin-C, fibrillin-1, and fibronectin in bullous keratopathy
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DOI：
10.1136/bjo.85.6.720
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发表时间：
2001-06-01
影响因子：
4.1
通讯作者：
Meek, KM
中科院分区：
文献类型：
--
作者：
Akhtar, S;Bron, AJ;Meek, KM
目的研究蛋白多糖（PG），β ig-h3（角膜上皮素），腱生蛋白-C（TN-C）），白蛋白，纤维连接蛋白在大泡性角膜病变（BK）corneas. Methods的超微结构定位5眼角膜从假晶状体大泡性角膜病变（BK）的情况下进行了检查，通过电子显微镜。PG被证明使用铜蛋白蓝，和蛋白β ig-h3，TN-C，白蛋白，和纤连蛋白的免疫定位与兔抗β ig-h3，小鼠抗TN-C（BC 10和TN 2），小鼠抗白蛋白-1（MAB 2502），小鼠抗白蛋白（MAB 1919），和兔抗纤连蛋白通过使用标准的免疫金technique.Results-Epithelial细胞含有大量空泡。存在上皮褶皱和大的电子透明上皮下大疱。基底层增厚，锚定丝断裂。在基质中，纤维间胶原间距增加，并存在异常大的PG。后弹力膜（DM.）包含了透明的空间，其中有小细丝。角质形成细胞和内皮细胞含有黑色素颗粒。后胶原层（PCL）含有大量的微丝和宽间距的胶原纤维，周期为100 nm。在PCL的内皮面观察到大量异常PG。在基底膜、Bowman层、基质、DM和PCL中观察到非常强的β ig-h3抗体标记，但在角膜细胞和内皮细胞中未观察到。BC 10和TN 2的强标记见于上皮下方、半桥粒缺失的电子透明间隙、纤维化血管翳、Bowman层、基质和Descemet膜的部分区域。用BC 10标记比用TN 2标记更强且分布更均匀。原纤蛋白-1（MAB 2502）和原纤蛋白（MAB 1919）标记与TN-C标记相似。Fibrillin（MAB 1919）的标记是强于tumorin-1（MAB 2502）labeling. Conclusions免疫电子显微镜显示在细胞和亚细胞水平的蛋白质的精确标记。与正常角膜相比，蛋白质β ig-h3、TN-C、纤维连接蛋白和纤维连接蛋白的表达高度增加。在水肿的间质中，增加的胶原纤维分离可能促进一些可溶性蛋白质如β ig-h3在整个间质中的更广泛分布。在这些BK病例中研究的蛋白质的修饰表达可被视为损伤反应的一部分。
Aims-To investigate the ultrastructural localisation of proteoglycans (PG), beta ig-h3 (keratoepithelin), tenascin-C (TN-C)), fibrillin, and fibronectin in bullous keratopathy (BK) corneas.Methods-Five corneas from cases of pseudophakic bullous keratopathy (BK) were examined by electron microscopy. PG were demonstrated using cuprolinic blue, and the proteins beta ig-h3, TN-C, fibrillin, and fibronectin were immunolocalised with rabbit anti-beta ig-h3, mouse anti-TN-C (BC10 and TN2), mouse antifibrillin-1 (MAB2502), mouse antifibrillin (MAB1919), and rabbit antifibronectin by using a standard immunogold technique.Results-Epithelial cells contained numerous vacuoles. Epithelial folds and large, electron lucent subepithelial bullae were present. Basal lamina was thickened and traversed by disrupted anchoring filaments. In the stroma, interfibrillar collagen spacing was increased and abnormally large PG were present. Descemet's membrane (DM.) contained lucent spaces in which there were small filaments. Keratocyte and endothelial cells contained melanin granules. A posterior collagenous layer (PCL) contained numerous microfilaments and wide spacing collagen fibres with a periodicity of 100 nm. Large quantities of abnormal PG were observed at the endothelial face of the PCL. Very strong labelling with beta ig-h3 antibody was observed in the basement membrane, Bowman's layer, stroma, DM, and PCL, but not in keratocytes and endothelial cells. Strong labelling with BC10 and TN2 was seen below the epithelium, in electron lucent spaces where the hemidesmosomes were absent, in the fibrotic pannus, in parts of Bowman's layer, the stroma, and Descemet's membrane. Labelling with BC10 was stronger and more evenly distributed than with TN2. Fibrillin-1 (MAB2502) and fibrillin (MAB1919) labelling was similar to TN-C labelling. Fibrillin (MAB1919) labelling was stronger than fibrillin-1 (MAB2502) labelling.Conclusions-Immunoelectron microscopy showed precise labelling of proteins at both the cellular and the subcellular level. Expression of proteins beta ig-h3, TN-C, fibrillin, and fibronectin was highly increased compared with normal cornea. In the oedematous stroma, increased collagen fibril separation may facilitate a wider distribution of some soluble proteins, such as beta ig-h3, throughout stroma. The modified expression of the proteins studied in these cases of BK may be regarded as part of an injury response.