Analysis of differential expression of glycosyltransferases in healing corneas by glycogene microarrays
Analysis of differential expression of glycosyltransferases in healing corneas by glycogene microarrays
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DOI：
10.1093/glycob/cwp133
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发表时间：
2010-01-01
期刊：
影响因子：
4.3
通讯作者：
Panjwani, Noorjahan
中科院分区：
文献类型：
--
作者：
Saravanan, Chandrassegar;Cao, Zhiyi;Panjwani, Noorjahan
一般认为，细胞表面上的聚糖和细胞外基质蛋白在介导伤口再上皮化的事件中起关键作用。然而，聚糖结构和组成的总体变化，特别是在角膜伤口闭合期间发生的变化仍然未知。本研究首次利用GLYCOv 2糖基因芯片技术鉴定小鼠角膜愈合过程中糖基化相关基因的差异表达。在阵列上的2000个糖基因中，与正常未损伤的角膜相比，愈合的角膜中11种糖基转移酶和糖苷酶的表达上调，19种下调超过1.5倍。其中，值得注意的是，糖基转移酶，β 3GalT 5，T-合成酶，和GnTIVb，都被发现在角膜中诱导响应损伤，而GnTIII和许多唾液酸转移酶下调。有趣的是，由于这些糖基转移酶的差异调节，在愈合的角膜中表达的糖蛋白和糖脂上的聚糖结构似乎可以作为半乳糖凝集素-3的特异性反受体，半乳糖凝集素-3是一种碳水化合物结合蛋白，已知在角膜伤口的上皮再形成中发挥关键作用。此外，首次发现许多糖基因（包括蛋白聚糖、磷脂酰肌醇蛋白聚糖-3、细胞粘附蛋白dectin-1和-2、mincle以及粘蛋白1）在角膜伤口愈合期间受到差异调节。通过qRT-PCR和凝集素印迹分析证实了糖基因微阵列数据的结果。在本研究中鉴定的差异表达的糖基因以前没有在伤口愈合的背景下进行过研究，并且代表了用于研究碳水化合物介导的识别在角膜伤口愈合中的作用的新因素。
It is generally accepted that the glycans on the cell surface and extracellular matrix proteins play a pivotal role in the events that mediate re-epithelialization of wounds. Yet, the global alteration in the structure and composition of glycans, specifically occuring during corneal wound closure remains unknown. In this study, GLYCOv2 glycogene microarray technology was used for the first time to identify the differentially expressed glycosylation-related genes in healing mouse corneas. Of similar to 2000 glycogenes on the array, the expression of 11 glycosytransferase and glycosidase enzymes was upregulated and that of 19 was downregulated more than 1.5-fold in healing corneas compared with the normal, uninjured corneas. Among them, notably, glycosyltransferases, beta 3GalT5, T-synthase, and GnTIVb, were all found to be induced in the corneas in response to injury, whereas, GnTIII and many sialyltransferases were downregulated. Interestingly, it appears that the glycan structures on glycoproteins and glycolipids, expressed in healing corneas as a result of differential regulation of these glycosyltransferases, may serve as specific counter-receptors for galectin-3, a carbohydrate-binding protein, known to play a key role in re-epithelialization of corneal wounds. Additionally, many glycogenes including a proteoglycan, glypican-3, cell adhesion proteins dectin-1 and -2, and mincle, and mucin 1 were identified for the first time to be differentially regulated during corneal wound healing. Results of glycogene microarray data were confirmed by qRT-PCR and lectin blot analyses. The differentially expressed glycogenes identified in the present study have not previously been investigated in the context of wound healing and represent novel factors for investigating the role of carbohydrate-mediated recognition in corneal wound healing.