Transgenic C.elegans as Amyloid Disease Model

描述(申请人提供)：我们已经设计了一种转基因秀丽线虫模型系统，允许在神经元或肌肉组织中诱导表达人β淀粉样多肽(Abeta1-42)。在这两种情况下，Abeta在细胞内积聚，形成淀粉样蛋白，并导致细胞病理。这项拟议工作的目标是了解这种毒性的分子和细胞基础，并调查这些过程可能如何参与阿尔茨海默病的病理。拟议的项目利用了线虫中可用的实验工具，其中许多工具在其他模型系统中是不存在的。具体地说，我们将使用正向基因筛查来识别抑制Abeta毒性的突变，从而识别与转导或对抗这种毒性有关的基因和基因产物。转基因动物还将用于基于微阵列的基因表达研究，从而能够从分子上描述细胞对Abeta毒性的初始反应。Abeta聚集和毒性之间的关系将通过构建更多表达天然和工程变异Abeta多肽的转基因株来进行研究。将使用基于双链RNA抑制(RNAi)和绿色荧光蛋白(GFP)的转基因报告构建来研究特定基因和基因产物在Abeta毒性中的作用。

DESCRIPTION (provided by applicant): We have engineered a transgenic Caenorhabditis elegans model system that allows inducible expression of human beta amyloid peptide (Abeta1-42) in neuronal or muscle tissue. In both instances, Abeta accumulates intracellularly, forms amyloid, and results in cellular pathology. The goal of the proposed work is to understand the molecular and cellular basis of this toxicity, and to investigate how these processes might be involved in Alzheimer's disease pathology. The proposed project takes advantage of the experimental tools available in C. elegans, many of which are not available in other model systems. Specifically, we will use forward genetic screens to identify mutations that suppress Abeta toxicity, thereby identifying genes and gene products involved in transducing or combating this toxicity. The transgenic animals will also be used in microarray-based gene expression studies, allowing a molecular description of the initial cellular response to Abeta toxicity. The relationship between Abeta aggregation and toxicity will be investigated by the construction of additional transgenic lines expressing natural and engineered variant Abeta peptides. The role of specific genes and gene products in Abeta toxicity will be investigated using double-stranded RNA inhibition (RNAi) and Green Fluorescent Protein (GFP)-based transgenic reporter constructs.