PHARMACOLOGY OF THE CORNEAL ENDOTHELIUM

内皮对于维持角膜透明度至关重要。 与 随着年龄的增长，人类角膜内皮细胞失去了分裂的能力。 以来 由于正常损耗，没有细胞损失的替代， 构成单层的细胞随时间减少。 在老年人中， 疾病、创伤或眼内手术的压力可进一步损害 这种组织，导致它失去了保持完整的能力， 单层，产生水肿，有时需要角膜 移植 我们的长期目标是开发药理学 治疗角膜内皮的年龄和疾病相关变化。 要做到这一点，我们必须发现角膜内皮细胞是如何对一种 单分子层中的缺陷通过破坏与它们的邻居的接触， 作为个体迁移以重新填充该地区，和/或集体迁移 扩大和变平，导致连续的单层细胞移动 或者扩散到缺陷区域中。 使用模拟的组织培养模型 成年人角膜内皮的行为，我们发现， 角膜内皮细胞迁移和扩散是不可分离的 运动的形式。 我们将使用这些模型：1）表征 区分迁移细胞和铺展细胞的形态学特征，2） 确定细胞外基质成分是否可能是内源性的 迁移或扩散的信号，3）识别细胞内 介导细胞迁移或扩散信号的途径，4） 确定基因表达和蛋白质中发生了什么样的修饰 合成、磷酸化和亚细胞组织， 迁移或扩散，以及5）识别特定的药理学试剂 刺激伤口修复 这些技术将用于 研究包括扫描电子显微镜，数字化形态测量， 和二维凝胶电泳，代谢标记， 免疫细胞化学定位，北方印迹分析，体外 翻译和原位杂交。 使用这些信息获得的信息 方法应该为设计药物治疗提供基础， 保护人类角膜内皮功能，刺激，也许， 加速内皮伤口愈合，并减少应力对 老化的内皮细胞 我们的研究还将建立角膜 内皮细胞作为单层组织创伤修复研究的模型 在眼睛和其他器官中。

The endothelium is critical for maintaining corneal transparency. With age, human corneal endothelial cells lose their ability to divide. Since there is no replacement of cells lost due to normal attrition the number of cells comprising the monolayer decreases over time. In older persons, the stress of disease, trauma, or intraocular surgery can further compromise this tissue, causing it to lose its ability to maintain an intact monolayer, producing edema and, sometimes, the need for corneal transplantation. Our long-term goal is to develop pharmacological treatment for age- and disease-related changes in the corneal endothelium. To do this, we must discover how corneal endothelial cells respond to a defect in the monolayer by breaking contact with their neighbors and migrating as individuals to repopulate the area, and/or by collectively enlarging and flattening, causing the continuous monolayer of cells to move or spread into the defect area. Using tissue culture models which mimic the behavior of adult human corneal endothelium, we have discovered that corneal endothelial migration and spreading are pharmacologically separable forms of movement. We will use these models to: 1) characterize the morphologic features which distinguish migrating from spreading cells, 2) determine whether extracellular matrix components might be endogenous signals for either migration or spreading, 3) identify the intracellular pathway which mediates the signal for cell migration or for spreading, 4) determine what modifications occur in gene expression and in protein synthesis, phosphorylation and subcellular organization to produce either migration or spreading, and 5) identify specific pharmacological agents which stimulate wound repair. Techniques that will be used in these studies include scanning electron microscopy, digitized morphometry, one and two-dimensional gel electrophoresis, metabolic labeling, immunocytochemical localization, Northern blot analysis, in vitro translation and in situ hybridization. Information obtained using these methods should provide a basis for designing drug treatments which will preserve human corneal endothelial function, stimulate and, perhaps, accelerate endothelial wound healing, and reduce the effects of stress on the aging endothelium. Our studies will also establish the corneal endothelium as a model for the study of wound repair in monolayer tissues in the eye and in other organs.