CELL SIGNAL TRANSDUCTION IN CORNEAL WOUND HEALING

描述：（改编自申请人摘要）角膜 创伤或手术后的上皮再生是恢复正常的基础 视野 PI计划检验以下假设： 磷脂酰肌醇-3-激酶（PI-3 K）和蛋白激酶C（PKC）是 在信号转导通路中的重要步骤导致修复。 这些 通路以级联方式将刺激从细胞表面传递到细胞核 有丝分裂原活化蛋白激酶（MAPK）。 的 角膜上皮层的清创术将用作伤口模型 治愈 PI建议测试PKC亚型是否可以直接激活 转录通过易位到细胞核或间接通过激活 MAPK级联，调节细胞增殖和伤口愈合。 她还 建议进行研究，以确定如何受体的表皮 表皮生长因子（EGF）和角质形成细胞生长因子（KGF）与 PI-3 K和定义哪些PKC亚型是PI-3 K的靶标。 选择性 抑制剂将用于将研究与伤口闭合相关联。 将使用HPLC和FPLC等分析方法。 鉴定 途径组分将通过特异性免疫印迹来完成。 抗体和激酶测定将在酶免疫复合物中进行 并通过磷光体成像定量。 获得的结果将定义 PI-3 K、PKC和MAPK参与角膜创伤愈合及辅助靶点 选择性调节角膜细胞信号转导的药物 上皮修复

DESCRIPTION: (adapted from the applicant s abstract) Corneal re-epithelization after trauma or surgery is fundamental to restore normal vision. The PI plans to test the hypothesis that phosphotidylinositol-3-kinase (PI-3K) and protein kinase C (PKC) are essential steps in the signal transduction pathway leading to repair. These pathways transmit stimuli from the cell surface to the nuclei in a cascade of events that involve mitogen-activated protein kinase (MAPK). The debridement of the corneal epithelial layer will be used as a model of wound healing. The PI proposes to test if PKC isoforms can directly activate transcription by translocating to the nuclei or indirectly by activating the MAPK cascade, regulating cell proliferation and wound healing. She also proposes to perform studies to establish how the receptors for epidermal growth factor (EGF) and keratinocyte growth factor (KGF) associate with PI-3K and to define which PKC isoforms are targets of PI-3K. Selective inhibitors will be used to correlate the studies with wound closure. Analytical procedures such as HPLC and FPLC will be used. Identification of pathway components will be accomplished by Western blot with specific antibodies, and kinase assays will be performed in enzyme immunocomplexes and quantified by phosphor imaging. The results obtained will define the involvement of PI-3K, PKC and MAPK in corneal wound healing and help target drugs to selective steps of signal transduction to modulate corneal epithelial repair.