CORNEAL EPITHELIAL DIFFERENTIATION AND REGENERATION

本项目的长期目标是了解 角膜发育、维护和再生。结果将 适用于更好地管理角膜愈合和预防 失明 研究策略是评估功能 特定蛋白质表达变化的意义， 在这些过程中， 我们关注的是一个Rho（Ras 同源性）-相关的丝氨酸/苏氨酸激酶（ROCK-I），其 在角膜上皮细胞中表达显著增加 在它们从角膜缘迁移到角膜表面之后。 Rho属于 Ras家族小GTP酶，其控制多种细胞 涉及细胞增殖、肌动蛋白丝重组的过程 和磷脂代谢。 ROCK-I是下游组件之一， Rho信号通路。 我们的近期目标是了解 ROCK-I参与角膜中信号级联的机制 上皮 为了实现这些目标，我们的具体目标是测试以下内容 假设：1）ROCK-1参与角膜上皮迁移， ROCK-I的这些功能 需要它的细胞内转运。 这将在体外进行测试 在培养的兔角膜上皮细胞中。2)磷酸化通过a cAMP依赖性蛋白激酶（PKA）参与控制 ROCK-1的酶活性和核转位。3)二酯 (DAG)，由受体触发水解形成的第二信使， 磷脂酰肌醇，4，5二磷酸（PIP 2），促进 ROCK-I的细胞内易位。 这些研究将导致 RhoA和ROCK-I信号传导元件的鉴定， 用于调节角膜上皮细胞的新药的特异性靶点 修复.

The long range goal of this project is to understand the mechanisms of corneal development, maintenance and regeneration. The results will be applicable to the better management of corneal healing and prevention of blindness. The research strategy is to evaluate the functional significance of changes in the expression of specific proteins which are regulated during these processes. We are focusing on a Rho (Ras homology)-associated in serine/threonine kinase (ROCK-I) whose expression is significantly increased in the corneal epithelial cells after they migrate from the limbal to corneal surface. Rho belongs to the Ras family of small GTPases, which control a variety of cellular processes involving cell proliferation, actin filament reorganization and phospholipid metabolism. ROCK-I is one of the downstream components of Rho signaling pathways. Our immediate goals are to understand the mechanisms of ROCK-I involvement in the signaling cascade in the corneal epithelium. To achieve these goals, our specific aims are to test the following hypotheses: 1) ROCK-I is involved in corneal epithelial migration, differentiation and proliferation and that these functions of ROCK-I require its intracellular translocation. This will be tested in vitro in cultured rabbit corneal epithelial cells. 2) Phosphorylation by a cAMP-dependent protein kinase (PKA) is involved in controlling the enzyme activity and nuclear translocation of ROCK-1. 3) diacylglycerol (DAG), a second messenger formed by receptor triggered hydrolysis of phosphotidyl inositol, 4, 5 biphosphate (PIP2), promotes the intracellular translocation of ROCK-I. These studies will lead to the identification of RhoA and ROCK-I signaling elements which may be specific targets for new drugs, useful for modulating corneal epithelial repair.