CHARACTERIZATION OF CORNEAL CELL-SURFACE ANTIGENS

角膜损伤或紊乱，包括角膜营养不良， 是视力受损和眼睛不适的常见原因。这个 细胞表面、细胞骨架与细胞周期的相互作用 细胞外基质(ECM)在伤口愈合过程中起着重要作用。一个 对这些相互作用机制的洞察将导致 为角膜问题设计更好的治疗方法。使用杂交瘤 技术，三种细胞骨架相关蛋白(CAP)是 被发现参与静止性角膜基质的激活 创面愈合过程中细胞从G1期到S期。建议数 研究将评估这些盖子的功能(一个相关 有肌动蛋白和两个有波形蛋白细丝)。 治愈。CAPS与肌动蛋白或肌动蛋白相互作用的机制 波形蛋白将在体外使用纯化的蛋白进行分析 通过研究它们在细胞中的分布模式 免疫电子显微镜。它们在GO过程中可能起的作用 GL向S的转变，将采用体外组织培养法进行研究 伤口愈合的模型。单细胞显微注射的效果 这些帽子和它们在细胞上的单抗 行为、细胞骨架组织和细胞功能 已评估。重组DNA技术将用于开发cdna 包含与这些基因的mRNAs互补的序列的克隆 并开发DNA和RNA探针。这些探测器将是 用来研究这些基因表达的调控 使用核酸杂交技术，在Will愈合过程中的帽子 技巧。 从以前的研究来看，基底膜和细胞表面 角膜上皮细胞中相关的独特蛋白质一直是 已确认身份。这种蛋白质被认为与细胞有关。 伤口愈合过程中的附着和迁移。它的作用机制 它在细胞-细胞和细胞-细胞外基质中的参与将通过 分析其与层粘连蛋白、硫酸乙酰肝素的相互作用 蛋白多糖、IV型胶原和纤维连接蛋白。监管机构 该蛋白在上皮愈合过程中的表达变化 将在体外伤口愈合的组织培养模型中进行研究 以及在兔体内的实验。重组DNA技术将用于 这项研究如CAPS所述。 这项研究的第三个方面将描述两个角膜 上皮细胞表面分化抗原 与角膜上皮愈合问题有关(复发 角膜上皮侵蚀与不完全性转分化 从结膜来源的细胞到角膜上皮细胞)。这个 生长因子对人脐静脉内皮细胞这些抗原表达的影响 培养的细胞及其生化特征将是 被执行。

Injury or disorders of the cornea, including corneal dystrophies, are common causes of impaired vision and eye discomfort. The interactions between the cell surface, cytoskeleton and the extracellular matrix (ECM) are important during wound healing. An insight into the mechanisms of these interactions will lead to devising better treatments for corneal problems. Using hybridoma techniques, three cytoskeletal associated proteins (CAPs) were found to be involved in the activation of quiescent corneal stromal cells from GO to Gl to S phase during wound healing. The proposed study will evaluate the functional of these CAPs (one associated with actin and two with vimentin filaments) in corneal wound healing. The mechanisms of interactions of the CAPs with actin or vimentin will be analyzed by using purified proteins in vitro and by studying the pattern of their distribution in cells using immunoelectron microscopy. Their possible functions during GO to Gl to S transition, will be studied in vitro using a tissue culture model of wound healing. The effects of single cell microinjections of these CAPs and their monoclonal antibodies on the cellular behavior, cytoskeletal organization and cellular functions will be evaluated. Recombinant DNA technology will be used to develop cDNA clones containing sequences complementary to the mRNAs for these CAPs and to develop DNA and RNA probes. These probes will be employed to study the regulation of the gene expression of theses CAPs during would healing, using nucleic acid hybridization techniques. From previous studies, a basement membrane and cell surface associated unique protein in corneal epithelial cells has been identified. This protein is postulated to be involved in cell attachment and migration during wound healing. The mechanisms of its involvement in cell-cell and cell-ECM will be studied by analyzing its interactions with laminin, heparan sulfate proteoglycan, type IV collagen and fibronectin. The regulatory changes in the expression of this protein during epithelial healing will be studied in tissue culture models of wound healing in vitro and in rabbit in vivo. Recombinant DNA techniques will be used for this study as described for CAPs. The third aspect of this study will characterize two corneal epithelial cell surface differentiation antigens which are associated with corneal epithelial healing problems (recurrent corneal epithelial erosion and incomplete transdifferentiation of conjunctivally derived cells to corneal epithelial cells). The effect of growth factors on the expression of these antigens in cultured cells and their biochemical characteristics will be carried out.