DEVELOPMENT AND MAINTENANCE OF LENS TRANSPARENCY

描述：透明的蜂窝结构是 生物镜头。接下来的五年申请将调查 晶状体细胞透明结构的分子基础。少年派的过去 对TC测量的集体相互作用的研究导致了临床 泛替辛作为抗白内障药物在人体上的试验。使用了泛替辛 因为它能有效对抗蛋白质聚集和晶状体混浊。 在动物模型中。在接下来的五年里， 选定的人类晶状体蛋白似乎参与了 相互作用对晶状体细胞的发育和维持很重要 透明度将受到调查。重组人晶状体蛋白 (野生型和突变型)将表达、纯化和鉴定 涉及人晶体蛋白相互作用的生化分析(AIM 1)。 傅立叶、幂定律和分形分析的新的定量方法将 用来表征晶状体细胞和模型的微观结构 人晶状体胞浆蛋白溶液(AIM 2)。方法将是 用于调查和量化专业的大小和顺序/无序 在晶状体细胞质中发现的结构成分，将评估分形性 晶状体细胞结构的尺寸。初步结果表明，阿尔法B 晶状体蛋白和细胞骨架蛋白在 发展和维护晶状体细胞的透明度。转基因和 白内障基因敲除模型将用于体内研究 晶状体晶体蛋白、细胞骨架蛋白与晶状体的关系 混浊(AIM 3)。拟议研究的结果将直接 在设计和测试新的治疗方法中的应用 抑制人类的蛋白质聚集和晶状体混浊。

DESCRIPTION: Transparent cellular structure is the fundamental property of the biological lens. The next five year application will investigate the molecular basis for the transparent structure of lens cells. The PI's past studies of the collective interactions measured by Tc led to a clinical trial of pantethine as an anticataract agent in humans. Pantethine was used because it was effective against protein aggregation and lens opacification in animal models. During the next five years, the functional motifs of selected human lens proteins that appear to be involved with the interactions important for development and maintenance of lens cell transparency will be investigated. Recombinant human lens crystallins (wild-type and mutant) will be expressed, purified and characterized using biochemical assays for interactions involving human crystallins (AIM 1). Novel quantitative methods for Fourier, power law and fractal analysis will be used to characterize the microscopic structure of lens cells and of model solutions of human lens cytoplasmic proteins (AIM 2). The methods will be used to investigate and quantify the size and order/disorder of major structural components found in lens cytoplasm and will evaluate the fractal dimension of lens cell structure. Preliminary results suggest that alpha B crystallin and cytoskeletal proteins have important functions in the development and maintenance of lens cell transparency. Transgenic and knockout models for cataract will be used for in vivo studies of the relationships between lens crystallins, cytoskeletal proteins and lens opacification (AIM 3). The results of the proposed studies will have direct application in the design and testing of new therapeutic approaches to inhibit protein aggregation and lens opacification in humans.