Functional Analysis of Novel Genes in Eye Development and Vision

眼睛发育和视力新基因的功能分析

• 批准号: 8737637
• 负责人: graeme j wistow
• 金额: $ 69.74万
• 依托单位: NATIONAL EYE INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8737637
• 关键词: Afferent Neurons; Age related macular degeneration; Aging; Animal Model; Binding; Bioinformatics; Carotenoids; Cell Culture Techniques; Cells; Cholesterol; Choroid; Complement Factor H; Complex; Defect; Deposition; Drug Design; Ear; Eye; Eye Development; Genes; Genomics; Genotype; Growth; Hearing; Human; Hybrids; Immunofluorescence Immunologic; Knockout Mice; Mass Spectrum Analysis; MicroRNAs; Modeling; Mus; Pathway interactions; Photoreceptors; Predisposition; Process; Protein Binding; Proteins; Recombinant Proteins; Retina; Retinal Cone; Retinoids; Role; Stress; Techniques; Tissues; Transgenic Mice; Variant; Vision; Visual; Yeasts; age related; disorder risk; mouse model; novel; promoter; protein complex; research study; response; retinal rods; risk variant

1: AMD and complement factor H (CFH). Common sequence variants of CFH have major roles in determining susceptibility to age-related macular degeneration (AMD). We have identified potentially important targets for CFH binding in human RPE/choroid. These results have been confirmed by immunofluorescence localization of multiple donor eyes and by using native and recombinant protein binding experiments and have been correlated with genotype for disease-risk variants of CFH. Protein complexes also appear to be associated with domains rich in cholesterol, another factor implicated in AMD. Mass spectroscopy has been used to identify further components of complexes that are implicated in formation of protein depositions in AMD. This has potential for drug-design to intervene in the progression of AMD. A cell-culture model for stress-related processes in AMD is also being investigated and shows that stressed RPE-derived cells up regulated genes for pathways important in AMD. 2: A miRNA cluster with high expression in eye and ear has been deleted in mouse, producing a new model of hearing and vision deficit. This has been examined in detail and provides evidence for involvement of miRNAs in maturation of photoreceptors, particularly of cone cells, and in other sensory neurons. 3: Retbindin is a novel protein of the retina. We have shown that retbindin is essentially specific for photoreceptors and that the human gene has separate cone and rod cell promoters. A transgenic mouse model shows suggests that retbindin has a role in control of retinoid and carotenoid levels in the retina. A knockout mouse has been created; it shows no defects in normal eye development and function but reveals age-related deficits in visual response

1：AMD和补体因子H（CFH）。 CFH的常见序列变异在确定与年龄相关的黄斑变性（AMD）的敏感性方面具有重要作用。我们已经确定了人类RPE/脉络膜中CFH结合的潜在重要目标。通过多个供体眼的免疫荧光定位以及使用天然和重组蛋白结合实验的免疫荧光定位证实了这些结果，并且与CFH疾病风险变体的基因型相关。蛋白质复合物似乎也与富含胆固醇的结构域有关，这是AMD的另一个因素。质谱学已用于鉴定与AMD中蛋白质沉积形成有关的复合物的进一步成分。这具有药物设计的潜力，可以干预AMD的进展。也正在研究AMD中应力相关过程的细胞培养模型，并表明应力的RPE衍生细胞超出了对AMD重要的途径的调节基因。 2：在小鼠中删除了具有高度表达的miRNA簇，产生了一种新的听力和视力不足模型。已经对此进行了详细研究，并为miRNA参与感光细胞的成熟，尤其是锥细胞和其他感觉神经元的成熟提供了证据。 3：视网膜是视网膜的一种新型蛋白质。我们已经表明，乙蛋白基本上是对感光体的特异性特异性，并且人类基因具有单独的锥体和杆细胞启动子。转基因小鼠模型表明，视网膜蛋白在视网膜类维生素类和类胡萝卜素水平的控制中起作用。已经创建了淘汰鼠标；它在正常的眼睛发育和功能中没有显示出缺陷，但揭示了与年龄相关的视觉反应缺陷