Functional Analysis of Novel Genes in Eye Development and Vision

1：AMD和补体因子H（CFH）。CFH的常见序列变体在确定年龄相关性黄斑变性（AMD）的易感性中具有主要作用。我们已经确定了CFH在人RPE/脉络膜中结合的潜在重要靶点。这些结果已被证实的免疫荧光定位的多个捐助者的眼睛，并通过使用本地和重组蛋白结合实验，并已与基因型的疾病风险变异CFH。蛋白质复合物似乎也与富含胆固醇的结构域相关，胆固醇是AMD中的另一个因素。质谱已被用于鉴定涉及AMD中蛋白质沉积形成的复合物的其他组分。这对药物设计干预AMD的进展具有潜力。AMD中应激相关过程的细胞培养模型也正在研究中，并显示应激RPE衍生细胞上调AMD中重要途径的基因。 2：在小鼠中删除了在眼和耳中高表达的miRNA簇，产生了新的听力和视力缺陷模型。这已被详细检查，并提供证据表明miRNA参与光感受器，特别是视锥细胞和其他感觉神经元的成熟。 Retbindin是一种新的视网膜蛋白。我们已经证明retbindin本质上是感光细胞特异性的，并且人类基因具有单独的视锥细胞和视杆细胞启动子。转基因小鼠模型显示，retbindin在控制视网膜中的类维生素A和类胡萝卜素水平中起作用。一只基因敲除小鼠已经被创造出来，它在正常的眼睛发育和功能上没有缺陷，但在视觉反应上显示出与年龄相关的缺陷。

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