Age-related Changes in Epithelial Microvilli

上皮微绒毛与年龄相关的变化

• 批准号: 8005877
• 负责人: VERA L BONILHA
• 金额: $ 11.07万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-07 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8005877
• 关键词: Acetylation; Actin-Binding Protein; Actins; Affect; Age; Aging; Aging-Related Process; Antibodies; Apical; Arts; Binding; Biochemical; Biological Assay; Buffers; Cell Aging; Cells; Cytoskeleton; Data; Development; Disease; Elderly; Enzymes; Epithelial Microvillus; Excision; Generations; Goals; Histocytochemistry; Homeostasis; Immunoelectron Microscopy; Immunoprecipitation; Length; Mediating; Membrane; Methodology; Methods; Methylation; Modification; Molecular; Molecular Profiling; Morphogenesis; Natural regeneration; Neural Retina; Nutrient; Phagocytosis; Phosphorylation; Phosphorylation Site; Photoreceptors; Physiological; Play; Post-Translational Protein Processing; Process; Proteins; Proteomics; Protocols documentation; Rattus; Research; Research Project Grants; Retina; Retinal Pigments; Role; Sampling; Signal Transduction; Signaling Molecule; Site; Staging; Structure; Structure of retinal pigment epithelium; Surface; Testing; Triton X100; Vision; Waste Products; age group; age related; aged; cellular microvillus; ezrin; innovation; insight; nitration; pathological aging; protein expression; protein function; receptor; research study; senescence; young adult

DESCRIPTION (provided by applicant): The establishment of an intimate interaction between the long retinal pigment epithelium (RPE) microvilli and the mature photoreceptor outer segments is essential for vision. The RPE performs highly specialized, unique functions essential for homeostasis of the neural retina. These include phagocytosis of photoreceptors shed outer segments, directional transport of nutrients into and removal of waste products from photoreceptor cells and visual pigment transport and regeneration. The apical microvilli of the RPE play a key role in mediating these activities. The hypothesis of this proposal is that proteomic comparison of young and old apical microvilli of the retinal pigment epithelium (RPE) will provide insight into fundamental RPE functions specifically affected by aging. This proposal explores the concept that the RPE apical microvilli can be studied to understand key age-related changes taking place in the RPE. Experiments in aim 1 will use a basic protocol to obtain intact RPE microvilli from young and aged rats. State-of-the-art proteomic methodologies and several biochemical assays will determine the protein composition and significant post- translational modifications associated with the samples in each age group. Confocal and immunoelectron microscopy analysis will further quantify the changes in the expression of relevant proteins. Experiments in aim 2 will characterize the role of ezrin in the aged RPE microvilli. Ezrin is an actin-binding protein involved in processes such as cell senescence and signal transduction. In the RPE, we have shown that ezrin is key to the development of apical microvilli and basal infoldings. This research will therefore also pursue a comparison of ezrin binding-partners in young and aged RPE apical microvilli and -MV fraction to better understand senescence-related changes in this cell. We will also investigate whether a differential post-translational modification of ezrin in aged RPE microvilli is somehow correlated to the age-related decrease of RPE apical microvilli. The long-term goal of this research is to identify proteins specifically affected by normal and pathological aging processes. The decline in vision in elderly, a direct consequence of changes in the neuro-retina and RPE, is a well-documented phenomenon. This research project will characterize physiological RPE ageing and relate it to ocular age-related diseases.

描述（由申请人提供）：长视网膜色素上皮（RPE）微绒毛和成熟感光器外节之间建立密切的相互作用对于视力至关重要。 RPE 执行高度专业化、独特的功能，对于神经视网膜的稳态至关重要。这些包括光感受器脱落外部节段的吞噬作用、营养物质定向输送到光感受器细胞和从光感受器细胞中去除废物以及视觉色素运输和再生。 RPE 的顶端微绒毛在调节这些活动中发挥着关键作用。该提案的假设是，对年轻和年老的视网膜色素上皮 (RPE) 顶端微绒毛进行蛋白质组学比较，将有助于深入了解受衰老影响的 RPE 基本功能。该提案探讨了可以研究 RPE 顶端微绒毛以了解 RPE 中发生的与年龄相关的关键变化的概念。目标 1 中的实验将使用基本方案从年轻和老年大鼠中获得完整的 RPE 微绒毛。最先进的蛋白质组学方法和多种生化测定将确定与每个年龄组的样品相关的蛋白质组成和显着的翻译后修饰。共聚焦和免疫电镜分析将进一步量化相关蛋白表达的变化。目标 2 中的实验将表征埃兹蛋白在老化 RPE 微绒毛中的作用。 Ezrin 是一种肌动蛋白结合蛋白，参与细胞衰老和信号转导等过程。在 RPE 中，我们表明埃兹蛋白是顶端微绒毛和基底内褶发育的关键。因此，本研究还将对年轻和老年 RPE 顶端微绒毛和 -MV 部分中的埃兹蛋白结合伴侣进行比较，以更好地了解该细胞中与衰老相关的变化。我们还将研究老年 RPE 微绒毛中埃兹蛋白的差异翻译后修饰是否与年龄相关的 RPE 顶端微绒毛减少有关。这项研究的长期目标是识别受正常和病理性衰老过程特别影响的蛋白质。老年人视力下降是神经视网膜和视网膜色素上皮变化的直接后果，这是一个有据可查的现象。该研究项目将描述生理性 RPE 衰老的特征，并将其与眼部年龄相关疾病联系起来。

项目成果

Loss of DJ-1 elicits retinal abnormalities, visual dysfunction, and increased oxidative stress in mice.

• DOI: 10.1016/j.exer.2015.07.014
• 发表时间: 2015-10
• 期刊: Experimental eye research
• 影响因子: 3.4
• 作者: Bonilha VL;Bell BA;Rayborn ME;Yang X;Kaul C;Grossman GH;Samuels IS;Hollyfield JG;Xie C;Cai H;Shadrach KG
• 通讯作者: Shadrach KG