TRANSGENIC C. ELEGANS AS AMYLOID DISEASE MODEL

转基因线虫作为淀粉样蛋白疾病模型

• 批准号: 6487825
• 负责人: Christopher D. Link
• 金额: $ 7.15万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-01 至 2003-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6487825
• 关键词: Alzheimer's disease; Caenorhabditis elegans; amyloid proteins; disease /disorder model; gene mutation; genetically modified animals; green fluorescent proteins; model design /development; molecular cloning; neurons; neurotoxins; recombinant proteins; suppressor mutations; western blottings

The goal of this project is to understand the cellular and molecular basis of beta-amyloid peptide (Abeta) toxicity. This peptide is deposited as insoluble amyloid aggregates in the senile plaques found in the brains of Alzheimer's Disease (AD) patients, and is believed to be central to the pathology of this disease. A novel AD model system will be employed in which of the intensely studied nematode worm, Caenorhabditis elegans, is engineered to express human Abeta peptide. These transgenic animals express high levels of the Abeta peptide, which rapidly leads to the formation of amyloid deposits and associated toxic effects. This model system will be used to investigate the relationships between Abeta structure, aggregation, and toxicity by the construction of transgenic animals expressing wild-type and variant Abeta containing single or multiple amino acid substitutions. These animals will be assayed for amyloid formation and known markers of Abeta toxicity, such as oxidative damage to proteins. Characterization of these animals will also include examination of the complete Abeta-dependent gene expression profile, assayed by mRNA hybridization to DNA microarrays that contain probes for the vast majority of C. elegans genes. The genetic accessibility of this model organism will also allow the identification of genes directly involved in Abeta toxicity, by the isolation and characterization of mutations that suppress the toxic effects of the Abeta peptide.

该项目的目标是了解β-淀粉样肽(Abeta)毒性的细胞和分子基础。这种多肽以不可溶的淀粉样蛋白聚集体的形式沉积在阿尔茨海默病(AD)患者大脑中发现的老年斑块中，被认为是这种疾病的病理中心。一种新的AD模型系统将被采用，在该系统中，被广泛研究的线虫--秀丽线虫--被设计成表达人的Aβ多肽。这些转基因动物表达高水平的Abeta多肽，这会迅速导致淀粉样蛋白沉积的形成和相关的毒性效应。该模型系统将通过构建表达含有单一或多个氨基酸取代的野生型和变异型Abeta的转基因动物来研究Abeta结构、聚集和毒性之间的关系。这些动物将被检测淀粉样蛋白的形成和已知的Abeta毒性标记物，例如对蛋白质的氧化损伤。这些动物的特征还将包括检查完整的依赖于Abeta的基因表达谱，通过与包含绝大多数线虫基因探针的DNA微阵列进行mRNA杂交来分析。这种模式生物的遗传可及性还将通过分离和表征抑制Abeta多肽毒性效应的突变来识别直接参与Abeta毒性的基因。