Unique distribution of thrombospondin-1 in human ocular surface epithelium
Unique distribution of thrombospondin-1 in human ocular surface epithelium
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DOI：
10.1167/iovs.05-1305
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发表时间：
2006-04-01
影响因子：
4.4
通讯作者：
Kinosbita, S
中科院分区：
文献类型：
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作者：
Sekiyama, E;Nakamura, T;Kinosbita, S
目的.本研究旨在阐明人眼表上皮中血管生成相关因子的详细表达模式。重点是角膜上皮（CE）基因表达显著高于结膜上皮（CJE）的因素。采用引入的扩增片段长度多态性（iAFLP）方法，比较了人CE和CJE中36种血管生成相关因子的相对基因表达。还研究了表达模式的CE，角膜缘上皮（LE），CJE的因素，显着更高的表达在CE中，通过使用实时PCR，原位杂交，免疫组化和免疫电镜。仅血小板反应蛋白（TSP）-1在CE中表现出显著更高的表达。原位杂交和实时PCR显示CE和LE的基底细胞中有TSP-1转录本。与CJE相比，它们在这些位点显著上调。免疫组化显示TSP-1在CE的基底部强表达。在LE中表达微弱，在CJE中不表达。免疫电子显微镜显示，CE和LE证明TSP-1标记上皮下方，在基底细胞的基底区域，偶尔在基底细胞膜。CJE中几乎没有或没有标记。在人眼表上皮中，CE和LE的基底细胞合成TSP-1，但CJE的基底细胞不合成TSP-1。高水平的TSP-1仅存在于CE以下。其独特的分布可能与角膜的无血供和完整性有关。
PURPOSE. The study was conducted to elucidate the detailed expression pattern of angiogenesis-related factors in human ocular surface epithelium. The focus was factors with significantly higher gene expression in corneal epithelium (CE) than in conjunctival epithelium (CJE).METHODS. The relative gene expression of 36 angiogenesis-related factors was compared in human CE and CJE, by using the introduced amplified fragment-length polymorphism (iAFLP) method. Also examined were the expression patterns in the CE, limbal epithelium (LE), and CJE of factors with significantly higher expression in the CE, by using real-time PCR, in situ hybridization, immunohistochemistry, and immunoelectron microscopy.RESULTS. Only thrombospondin (TSP)-1 exhibited significantly higher expression in the CE. In situ hybridization and real-time PCR showed TSP-1 transcripts in the basal cells of the CE and LE. Compared with the CJE, they were significantly upregulated at those sites. Immunohistochemistry revealed that TSP-1 was strongly expressed in the basal region of the CE. Its expression was faint in LE and absent in CJE. Immunoelectron microscopy revealed that the CE and LE demonstrated TSP-1 labeling just below the epithelium, in the basal region of basal cells, and occasionally in the basal cell membrane. There was little or no labeling in the CJE.CONCLUSIONS. In the human ocular surface epithelium, basal cells of the CE and LE, but not of the CJE, synthesize TSP-1. High levels of TSP-1 are present only just below the CE. Its unique distribution may be related to corneal avascularity and integrity.