The developmental proteome of retinal progenitor

视网膜祖细胞的发育蛋白质组

• 批准号: 6779921
• 负责人: MARY Heather WEST GREENLEE
• 金额: $ 14.37万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6779921
• 关键词: cell differentiation; cell membrane; developmental genetics; laboratory mouse; matrix assisted laser desorption ionization; membrane proteins; neurogenesis; protein purification; protein sequence; protein structure function; proteomics; retina; retina disorder; cell differentiation; cell membrane; developmental genetics; laboratory mouse; matrix assisted laser desorption ionization; membrane proteins; neurogenesis; protein purification; protein sequence; protein structure function; proteomics; retina; retina disorder

DESCRIPTION (provided by applicant): Retinal dystrophies are diseases that involve degeneration of photoreceptors, the sole light-sensing cells in the retina. A solid understanding of retinal development can contribute to new therapies for combating retinal dystrophies. Recent advances in stem-cell research may make replacement of retinal neurons a feasible therapy for the treatment of many retinal degenerative disorders. In this regard there is currently only a poor understanding of the components of a progenitor cell/stem cell that change as it becomes a particular type of retinal neuron. The purpose of the proposed studies is to apply the powerful techniques of proteomics as a novel research tool in the study of retinal development. Because the ability of a progenitor cell to respond to cues from its environment are reflected in the proteins present in the 'proteome' of the plasma membrane, this project will investigate changes in expression among plasma membrane proteins of retinal progenitor cells from 3 developmental ages. The specific aims of the proposal are (1) To isolate retinal progenitor cells from the developing retina (embryonic days 15 and 18, postnatal day 0), and purify and separate proteins of the plasma membrane. (2) To catalog the proteins of the progenitor cell plasma membrane at these developmental ages, and quantify their levels of expression. Proteins will be identified by using matrix assisted laser desorption ionization-time of flight mass spectrometry, and using generated spectra to search against protein sequence databases. (3) To cluster proteins into theoretical functional groups based on how their level of protein expression changes. This study will be the first to apply the tools of proteomics to retinal development and will pave the way for future developmental studies using this powerful technique. Further, this analysis will identify dozens of proteins and signaling pathways of importance for retinal development, and help to more efficiently focus further studies of differentiation of retinal progenitors.

描述（由申请人提供）：视网膜营养不良是涉及视网膜中唯一的光感应细胞的感光体变性的疾病。 对视网膜发育的扎实理解可以为打击视网膜营养不良的新疗法做出贡献。 干细胞研究的最新进展可能会使视网膜神经元替换成为许多视网膜退行性疾病的可行疗法。在这方面，目前只有对祖细胞/干细胞的成分的理解不足，因为祖细胞/干细胞变成了特定类型的视网膜神经元。 拟议的研究的目的是将蛋白质组学的强大技术应用于视网膜发育研究中的新型研究工具。 由于祖细胞对环境提示的反应能力反映在质膜“蛋白质组”中的蛋白质中，因此该项目将研究来自3个发育年龄的视网膜祖细胞的质膜蛋白之间表达的变化。 该提案的具体目的是（1）将视网膜祖细胞与发育中的视网膜分离（胚胎第15天和第18天，产后第0天），并纯化和分离质膜的蛋白质。 （2）分类这些发育年龄的祖细胞质膜的蛋白质，并量化其表达水平。 将通过使用飞行质谱法的基质辅助激光解吸时间来鉴定蛋白质，并使用生成的光谱来搜索蛋白质序列数据库。 （3）基于其蛋白质表达水平的变化，将蛋白聚类到理论官能团中。 这项研究将是第一个将蛋白质组学工具应用于视网膜开发的工具，并将使用这种强大的技术为未来的发展研究铺平道路。 此外，该分析将确定数十种蛋白质和对视网膜发育重要性的信号传导途径，并有助于更有效地集中于视网膜祖细胞分化的进一步研究。