SPECTROSCOPY ON LENS MEMBRANE LIPID

我们提议的调查的总体目标是使用 最先进的光谱技术来检查区域 的构象、组成和结构特征 晶状体膜脂和蛋白质。光谱发现可能会 与膜的组成和性能有关。我们会 研究正常牛和人的晶状体，白内障人 镜片和镜片经过各种物理化学处理 压力。这样的研究很好地允许我们开发一个深入的 评价膜性质的相对微妙变化 这可能会导致膜性能的显著变化 会导致白内障的发生。这项研究的结果将 巩固确保未来所需的知识基础 人类白内障的医学治疗进展。 这项研究的具体目的如下： 1.测量初级红外光谱和拉曼光谱 正常人脂泡分散体的特性研究 牛和人类的镜片。光谱特征将 包括：a)流动性和相变温度；b) 正交型或单斜型堆积顺序；c)脂头基 相互作用；d)脂酰基-界面区有序性和堆积； E)定量脂类饱和度；f)确认脂类成分。 囊泡分散体的膜脂成分为 用薄层层析法和高效 液体色谱学。 2.确定内膜和外膜的影响 蛋白质对晶状体细胞光谱特性的影响 膜。 3.探索伴随而来的光谱特征变化 晶状体和人类白内障的老化。 4.确定氧化应激、紫外线照射、 钙和胆固醇结合的抗生素非利平对心脏的影响 晶状体细胞膜和膜的光谱特性 脂类。 5.探讨膜脂光谱的相关性 与晶状体膜性能有关的特性，即ATPase 活性和膜泡溶质通量。

The overall objective of our proposed investigation is to use state-of-the-art spectroscopic techniques to examine regional conformational; compositional and structural characteristics of lens membrane lipids and proteins. Spectroscopic findings could be related to membrane composition and performance. We will investigate normal bovine and human lenses, cataractous human lenses and lenses subjected to a variety of physicochemical stresses. Such studies well permit us to develop an in depth appreciation of relatively subtle changes in membrane properties which can lead to considerable changes in membrane performance contributing to cataractogenesis. The results of this study will strengthen the foundation of knowledge necessary to assure future development of medical therapy of human cataract. The specific aims of this study are as follows: 1. Measure primary infrared and Raman spectroscopic characteristics of lipid vesicle dispersions obtained from normal bovine and human lenses. Spectroscopic characteristics will include: a) fluidity and phase transition temperatures; b) orthorhombic or monoclinic packing order; c) lipid head group interactions; d) lipid acyl-interface region order and packing; e) quantitate lipid saturation; f) confirm lipid composition. The membrane lipid composition of the vesicle dispersions will be determine by thin layer chromatography and high performance liquid chromatography. 2. Determine the influence of intrinsic and extrinsic membrane proteins upon spectroscopic characteristics of lens cell membranes. 3. Explore changes in spectroscopic characteristics accompanying aging of the lens and in human cataract. 4. Determine the impact of oxidative stress, UV light exposure, calcium and the cholesterol binding antibiotic filipin upon the spectroscopic characteristics of lens cell membranes and membrane lipids. 5. Explore the correlation of membrane lipid spectroscopic characteristics with lens membrane performance i.e. ATPase activity and membrane vesicle solute flux.