ANALYSIS OF MICRODISSECTED CATARACTOUS HUMAN LENSES

这项研究项目的长期目标是了解 导致人类白内障形成的原因和机制。 具体目标是：(1)启动长期临床生物化学 与年龄和药物相关的人类白内障的调查。在这些 研究后，将对患者进行全面的术前评估 已执行。将观察患者的白内障进展情况。 临床上，并将使用标准化的裂隙灯程序进行记录， 偏振逆光照相和Scheimpflg照相。 白内障手术后，将对晶状体进行显微解剖和评估。 根据目标2和3描述的生物化学。(2)显微解剖单个 人白内障晶状体显微镜下的分离 鸦片疫区和邻近的正常地区。高性能液体 将使用色层分析(Hplc)来研究水的性质。 可溶性蛋白质、尿素可溶性蛋白质和两者中的膜蛋白 清晰和不透明的部分。其他技术将包括 十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、等电聚焦、氨基酸 放射性碘标记蛋白质组分的分析、胰酶多肽图谱 采用双向电泳层析系统 然后是放射自显影。(3)分析主要的非蛋白低谷。 人体不透明和透明切片的分子量组成 隐形眼镜。(4)开发取回镜片材料的新方法 囊外手术。要采取的接近将是敲击吸气线 手术过程中使用的设备。(5)识别和描述 23,000道尔顿晶体蛋白在许多情况下被选择性降解和修饰 人白内障晶状体蛋白水解酶的鉴定与研究 它们在一定程度上参与了这一过程。(6)完成 晶状体质膜主干基因克隆的核苷酸序列分析 内源蛋白(MIP-26)。(7)完成该病毒的核苷酸分析 牛β23/26晶状体蛋白。(8)研究饮食和衰老对健康的影响 小鼠γ-晶状体蛋白的差异性丢失 卡路里饮食。

The long term objectives of this research project are to understand the causes and mechanisms which lead to the formation of cataract in humans. The specific aims are: (1) To start a long term clinicobiochemical investigation of aging-related and drug-related human cataracts. In these studies, a comprehensive presurgical evaluation of the patient will be performed. The progression of cataract in the patient will be followed clinically, and will be documented using standardized slit lamp procedures, polarized retroillumination photography, and Scheimpflug photography. After cataract surgery, the lens will be microdissected and evaluated biochemically as described under aims 2 and 3. (2) To microdissect single human cataractous lenses and to separate with the aid of a microscope opague areas and adjacent normal areas. High performance liquid chromatography (HPLC) will be used to study the properties of the water soluble proteins, urea soluble proteins, and the membrane proteins in both the clear and opaque sections. Other techniques will include SDS-polyacrylamide gel electrophoresis, isoelectric focusing, amino acid analysis, tryptic peptide mapping of radioiodinated protein fractions employing the two-dimensional system of electrophoresis and chromatography followed by autoradiography. (3) To analyze the major nonprotein low molecular weight constituents in opaque and clear sections of human lenses. (4) To develop new methods for retrieving lens material during extracapsular surgery. To approach taken will be tap the aspiration lines of the equipment used during surgery. (5) To identify and characterize a 23,000 dalton crystallin which is selectively degraded and modified in many human cataractous lenses, and to identify and study the proteolytic enzymes which are involved, in part, in this process. (6) To complete the nucleotide sequencing of cDNA clones of the lens plasma membrane main intrinsic protein (MIP-26). (7) To complete the nucleotide analysis of the cow beta 23/26 crystallin. (8) To study the effects of diet and aging on the differential loss of gamma-crystallin in mice which are given a reduced calorie diet.