MICRODISSECTED CATARACTOUS LENSES

描述(申请人摘要)：本项目的长期目标 建议更好地了解生物化学和生物物理变化 发生在人类晶状体中，随着年龄的增长，最终会导致 白内障。拟议的研究将有助于在以下方面实现 未来，最终目标是显著延缓 老年性白内障以及消除继发性白内障的形成 白内障是白内障手术后常见的一种疾病。 特定目标I：鉴定人晶状体上皮蛋白：关键 维持晶状体动态平衡的关键是上皮层。 白内障后将获得的新鲜切除的人上皮 手术将会被使用。测量将在个人身上进行 上皮组织。上皮的一部分将用于酶活性 确定，而剩余的部分将用于确定 用聚合酶链式反应技术检测特定酶的mRNA水平。这个 人上皮细胞cDNA文库的构建将使我们能够表征 各种人类上皮蛋白，并找出是否有晶状体特有的 任何一种蛋白质的异构体。 特殊目的II：阐明新发现的分子伴侣 α-晶体蛋白的功能：分子伴侣是非常有效的 抑制变性蛋白质的聚集，这是老化晶状体的一个特性 需要。我们发现α-晶状体蛋白是一种分子伴侣。这 这表明它在晶状体中具有双重作用。一种作为折射元件 第二个作为伴侣，保护其他蛋白质不会聚集。 α-晶体蛋白的物理性质将通过不同的方法进行研究 技巧。α-晶状体蛋白与其他晶状体蛋白的相互作用 例如Beta、Gamma和MP-26将被评估。老龄化的影响和 关于α-晶体蛋白伴侣功能的白内障将在 人类正常晶状体和白内障晶状体。

DESCRIPTION (Applicant's abstract): The long term objective of this proposal is to better understand the biochemical and biophysical changes that occur in the human lens with aging, and which ultimately lead to cataract. The proposed studies will be useful for achieving, in the future, the ultimate goal of significantly retarding the formation of aging-dependent cataract as well as eliminating the formation of secondary cataract which is commonly encountered following cataract surgery. Specific Aim I: To characterize human lens eptithelial proteins: A key player in the maintenance of lens homeostasis is the epithelial layer. Freshly excised human epithelium which will be obtained following cataract surgery will be used. Measurements will be performed on individual epithelium. A portion of the epithelium will be use for enzyme activity determination, whereas the remaining piece will be used for determination of mRNA levels of the specific enzyme using PCR technology. The construction of human epithelial cDNA library will allow us to characterize various human epithelial proteins, and to find if there are lens- specific isoforms for any of the proteins. Specific Aim II: To elucidate the newly discovered molecular chaperone function of alpha-crystallins: Molecular chaperones are very efficient in suppressing aggregation of denatured protein, a property the aging lens needs. Alpha-crystallin was found by us to be a molecular chaperone. This suggests that it has a dual role in the lens. One as a refractive element and a second one as a chaperone protecting other proteins from aggregation. The physical properties of alpha-crystallins will be studied by various techniques. The interaction of alpha-crystallin with other lens proteins such as beta, gamma, and MP-26 will be evaluated. The effects of aging and cataract on the chaperone function of alpha-crystallin will be examined in normal and cataractous human lenses.