Ocular proteomics of retina

视网膜的眼部蛋白质组学

• 批准号: 6620900
• 负责人: HIROYUKI MATSUMOTO
• 金额: $ 36.55万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6620900
• 关键词: Internet; SDS polyacrylamide gel electrophoresis; immunocytochemistry; laboratory mouse; laboratory rat; mass spectrometry; matrix assisted laser desorption ionization; molecular biology information system; peptide library; protein sequence; protein structure; proteomics; radiotracer; retina; structural biology; vision

DESCRIPTION (From the Applicant's Abstract): The long aboutterm goal of this project is to define all the proteins expressed in rodent retinas in order to provide vision researchers with information regarding the proteins actually expressed in retinal cells. Protein analysis by the combination of two-dimensional (2-D) gel electrophoresis, mass spectrometry, and genome database search has impacted on the progress of biomedical sciences. This new breed of technology is generally called "proteomics" and enables massive analysis of proteins extracted from cells or tissues. In the past two decades the understanding of visual transduction pathway and its underlying molecular mechanisms has substantially been improved by the use of rod outer segment preparations from various animal models including bovine, frog, and other animals. The recent transgenic mouse model also has substantiated and extended much of the previous knowledge on the molecular mechanisms underlying vision. In this proposal, a proteomic approach will be used to characterize the proteins expressed in the retinas of laboratory rodents, rats and mice. The reasons for the choice of rodents as a model to study retinal protein expression are three-fold. First is the similarity in gene sequence between human and rodents. Second is the availability of transgenic animal models and the accumulated literatures on the physiology and pathological changes of rodent retinas induced by environmental or genetic perturbation. Finally, the lower costs and ease of experimental manipulation makes a rodent model practical. There are three specific aims for this proposal. Aim 1 is to profile virtually all the retinal proteins that are extracted from rodent retinas and separated on 2-D gels. Protein profiling will be made along the developmental time axis by investigating retinas at different postnatal days. In this aim, efforts will also be made to develop a technique to remove abundant proteins from the protein extract in order to investigate proteins expressed in minor quantities, and also to develop a mass spectrometric technique to quantify proteins by stable isotope labeling. Aim 2 is to profile immunohistochemical localization of selected proteins identified in Aim 1. In this aim, a defined set of criteria will be used so that the proteins chosen will be classified into groups that are physiologically distinct. Aim 3 is to make the proteomic information accessible on a web-based database so that vision researchers can retrieve data when needed. Accomplishment of this project will help researchers investigating vision and its disorders at the protein level using rodent models.

描述（来自申请人的摘要）： 这个项目的长期目标是确定所有表达的蛋白质 在啮齿动物视网膜中，以便为视觉研究人员提供信息 关于视网膜细胞中实际表达的蛋白质。蛋白质分析 二维（2-D）凝胶电泳、质谱 光谱学和基因组数据库搜索影响了 生物医学科学这种新技术通常被称为 “蛋白质组学”，使大量分析蛋白质提取的细胞或 组织中在过去的二十年里，对视觉信号转导的理解 途径及其潜在的分子机制已经得到了实质性的改善 通过使用来自各种动物模型的杆外节制备物 包括牛、青蛙和其他动物。最近的转基因小鼠模型 也证实和扩展了许多以前的知识， 视觉的分子机制在这项提议中，蛋白质组学方法 将用于表征在视网膜中表达的蛋白质， 实验室啮齿动物，大鼠和小鼠。选择啮齿类动物作为 研究视网膜蛋白表达的模型是三重的。首先是 人类和啮齿类动物基因序列相似性。二是 转基因动物模型的可用性和积累的文献 环境因素对啮齿类动物视网膜生理和病理的影响 或者基因干扰最后，成本较低，易于实验 操作使得啮齿动物模型实用。有三个具体目标， 这个提议。目的1是分析几乎所有的视网膜蛋白， 从啮齿动物视网膜中提取并在2-D凝胶上分离。蛋白质谱将 沿着发育时间轴，通过研究视网膜在不同的 产后的日子为此，还将努力开发一种技术， 从蛋白质提取物中除去大量蛋白质， 少量表达的蛋白质，并且还产生大量的 通过稳定同位素标记来定量蛋白质的光谱技术。目的2 是对所选蛋白质的免疫组织化学定位进行分析 目标1。为此，将使用一套确定的标准， 选择的蛋白质将被分类为生理上 与众不同目标3是使蛋白质组学信息可以在基于网络的 数据库，以便视觉研究人员可以在需要时检索数据。 该项目的完成将有助于研究人员研究视觉， 在蛋白质水平上使用啮齿动物模型来研究其紊乱。