Mutational Analyses of Drosophila DJ-1 Homologs

果蝇 DJ-1 同源物的突变分析

• 批准号: 6864841
• 负责人: Leo J Pallanck
• 金额: $ 17.63万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6864841
• 关键词: Drosophilidae; Parkinson' s disease; RNA interference; RNase protection assay; biotechnology; density gradient ultracentrifugation; disease /disorder model; dopamine; electrophysiology; functional /structural genomics; gene expression; gene mutation; gene targeting; genetic screening; immunocytochemistry; laboratory rabbit; microarray technology; model design /development; neural degeneration; neurons; neuropathology; nucleic acid structure; pathologic process; transposon /insertion element; western blottings

DESCRIPTION (provided by applicant): Parkinson's disease is a prevalent neurodegenerative disorder characterized by tremors, rigidity, and bradykinesia. These symptoms arise from the degeneration of dopaminergic neurons in the substantia nigra. The cellular and molecular mechanisms responsible for neurodegeneration in Parkinson's disease remain poorly understood, although genetic and environmental factors both appear to play contributing roles. Recently, loss-of-function mutations in DJ-1, a gene of unknown function, were found to be responsible for an autosomal recessive form of Parkinson's disease. To explore the normal biological function of DJ-1, and the mechanism by which loss of DJ-1 function results in neurodegeneration, we propose to subject a pair of highly conserved Drosophila DJ-1 homologs (designated DJ-1a and DJ-1b) to mutational analysis. DJ-1a and DJ-1b function will be perturbed using P element mutagenesis, gene-targeting and double stranded RNA interference methods. The phenotypes resulting from perturbation of these genes will be fully characterized, including an analysis of dopaminergic neuron integrity. Additionally, we will characterize the global gene expression changes resulting from loss of DJ-1a and DJ-1b function and initiate screens for genetic modifiers of the DJ-1a and DJ-1b phenotypes to elucidate the biochemical pathways in which these genes function. This work should clarify the normal cellular role of DJ-1 and provide a foundation for further hypothesis-driven investigation of DJ-1 function.

描述（由申请人提供）：帕金森病是一种常见的神经退行性疾病，其特征为震颤、强直和运动迟缓。这些症状是由黑质中多巴胺能神经元的退化引起的。尽管遗传和环境因素似乎都发挥了作用，但对帕金森病神经变性的细胞和分子机制仍知之甚少。最近，功能未知的基因 DJ-1 的功能丧失突变被发现与常染色体隐性遗传的帕金森病有关。为了探索 DJ-1 的正常生物学功能，以及 DJ-1 功能丧失导致神经变性的机制，我们建议对一对高度保守的果蝇 DJ-1 同源物（称为 DJ-1a 和 DJ-1b）进行研究。到突变分析。 DJ-1a 和 DJ-1b 功能将通过 P 元件诱变、基因靶向和双链 RNA 干扰方法受到干扰。这些基因扰动所产生的表型将得到充分表征，包括对多巴胺能神经元完整性的分析。此外，我们将描述由于 DJ-1a 和 DJ-1b 功能丧失而导致的全局基因表达变化，并启动 DJ-1a 和 DJ-1b 表型遗传修饰剂的筛选，以阐明这些基因发挥作用的生化途径。这项工作应阐明 DJ-1 的正常细胞作用，并为进一步假设驱动的 DJ-1 功能研究奠定基础。

项目成果

Drosophila DJ-1 mutants are selectively sensitive to environmental toxins associated with Parkinson's disease.

果蝇 DJ-1 突变体对与帕金森病相关的环境毒素选择性敏感。

• 发表时间: 2005-09-06
• 作者: Meulener, Marc;Whitworth, Alexander J;Armstrong;Rizzu, Patrizia;Heutink, Peter;Wes, Paul D;Pallanck, Leo J;Bonini, Nancy M
• 通讯作者: Bonini, Nancy M