CORNEAL EPITHELIAL CELL SURFACE GLYCOCONJUGATES

描述(改编自申请人的摘要)：过去的研究 申请人已证明新乳糖鞘糖脂(Neolacto-GSLs) 角膜上皮是调节角膜的关键分子之一。 上皮性伤口的体外封闭。目前的项目建议确定 Neolacto-GSLS调节角膜上皮片的机制 体外迁移使用兔模型来确定是否 新生Lacto-GSLs介导伤口再上皮化 在活体内。使用细胞-底物黏附试验，涉及黏附的试验 整合素脂质体对多种细胞外基质分子的影响及Western 对磷酸化蛋白质的印迹分析，申请人将尝试 建立角膜上皮细胞表面新生乳酸-GSLS 调节：细胞-底物相互作用(目标1)，Beta1的功能 整合素受体(AIM 2)与酪氨酸磷酸化介导的信号转导 路径(目标3)。使用体外角膜上皮伤口愈合试验， 申请者将确定新乳酸型GSLS是否促进角膜 与一种或多种内源性凝集素相互作用闭合上皮伤口 在细胞基质中(目标4)。为了评估体外试验的影响 体内愈合的结果(目标5)，申请者将确定是否 外源性Neolacto-GSLs促进擦伤和浅表创面愈合 活体兔角膜切削伤的实验研究。

DESCRIPTION (Adapted from applicant's abstract): Past studies by the applicant have shown that neolacto-glycosphingolipids (neolacto-GSLs) of the corneal epithelium are among the key molecules that mediate corneal epithelial wound closure in vitro. The current project proposes to identify the mechanism by which the neolacto-GSLs mediate corneal epithelial sheet migration in vitro using a rabbit model to determine whether the neolacto-GSLs of corneal epithelium mediate re-epithelialization of wounds in vivo. Using cell-substratum adhesion assays, assays involving adhesion of integrin-liposomes to various extracellular matrix molecules and Western blot analysis of phosphorylated proteins, the applicant will attempt to establish whether the corneal epithelial cell surface neolacto-GSLs modulate: cell-substratum interactions (aim 1), the function of beta1 integrin receptors (aim 2) and tyrosine phosphorylation-mediated signalling pathways (aim 3). Using in vitro corneal epithelial wound healing assays, the applicant will determine whether the neolacto-GSLs promote corneal epithelial wound closure by interacting with one or more endogenous lectins in the cell substratum (aim 4). To evaluate the impact of the in vitro findings to healing in vivo (aim 5), the applicant will determine whether the exogenous neolacto-GSLs promote healing of the scrape and superficial keratectomy wounds of rabbit corneas in vivo.