Use of C.elegans to Explore Bacterial Sources of Toxicity in Parkinson's Disease

利用秀丽隐杆线虫探索帕金森病毒性的细菌来源

• 批准号: 7021515
• 负责人: Guy A Caldwell
• 金额: $ 13.01万
• 依托单位: UNIVERSITY OF ALABAMA IN TUSCALOOSA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7021515
• 关键词: Caenorhabditis elegans; Parkinson' s disease; RNA interference; bacterial toxins; bioassay; disease /disorder etiology; disease /disorder model; dopamine; environmental toxicology; fluorescent dye /probe; gene environment interaction; gene expression profiling; genetic susceptibility; genetically modified animals; model design /development; protease inhibitor; proteasome; toxicant screening

DESCRIPTION (provided by applicant): Parkinson's disease (PD) results from an imbalance in cellular mechanisms designed to cope with environmental stresses to neurons. The two major clinical hallmarks of PD, protein inclusions termed Lewy bodies and dopamine neuron degeneration, are representative of failure in the intracellular management of stress. While genetic forms of PD are rare, these mutations highlight the involvement of pathways that regulate protein folding and oxidative damage in cells. Given the predominance of sporadic PD, environmental sources of toxins may serve as potential risk factors for individuals with specific genetic predispositions. One environmental factor that may contribute to PD is exposure to certain bacteria that produce proteasome inhibitors, such as specific strains of the order Actinomycetales. Here we propose to utilize the nematode roundworm, C. elegans, to mechanistically investigate exposure to bacterial strains implicated in PD. Our lab has previously established this worm model for rapid evaluation of factors influencing both the misfolding of human alpha-synuclein and neuroprotection of dopamine neurons. The aims of our proposal include investigating dopamine neuron degeneration as caused by bacterial exposure in wild-type worms and in genetically defined backgrounds. We will evaluate susceptibility to bacterial exposure in animals defective in worm homologs of known PD genes, in addition to novel PD gene targets obtained from a large-scale RNA interference (RNAi) screen we have performed. Transgenic nematodes containing fluorescent reporter gene constructs will be used to distinguish systemic effects of exposure on various neuronal subtypes, general stress response, and proteasomal inhibition. Differential changes in gene expression in response to bacterial exposure will also be profiled using whole-genome oligonucleotide microarrays to identify potential genes regulated in response to environmental toxins. Relevance to Public Health: The interplay between genetic predisposition and susceptibility to environmental insults lies at the core of PD. Risk factors are best evaluated using systems wherein environmental conditions and genetic differences are strictly controlled. C. elegans, a microscopic worm with precisely 8 dopamine neurons, shares about half of its genes with humans and represents an ideal system to rapidly examine potential sources environmental toxins that may influence development of PD.

描述(由申请人提供)：帕金森氏病(PD)是由旨在应对环境压力对神经元的细胞机制的不平衡造成的。帕金森病的两个主要临床特征，称为路易小体的蛋白质包涵体和多巴胺神经元变性，代表了细胞内应激管理的失败。虽然帕金森病的遗传形式很少见，但这些突变突显了调节蛋白质折叠和细胞氧化损伤的途径的参与。鉴于散发性帕金森病的优势，环境中的毒素来源可能是具有特定遗传倾向的个体的潜在危险因素。可能导致帕金森病的一个环境因素是接触到某些产生蛋白酶体抑制剂的细菌，例如放线菌目的特定菌株。在这里，我们建议利用线虫线虫，秀丽隐杆线虫，机械地研究与帕金森病有关的细菌菌株的暴露。我们的实验室之前已经建立了这个蠕虫模型，用于快速评估影响人类α-突触核蛋白错误折叠和多巴胺神经元神经保护的因素。我们建议的目的包括调查由细菌暴露在野生型蠕虫和遗传背景下引起的多巴胺神经元退化。除了从我们进行的大规模RNA干扰(RNAi)筛选中获得的新的PD基因靶点外，我们还将评估在已知PD基因的蠕虫同源物中存在缺陷的动物对细菌暴露的敏感性。含有荧光报告基因结构的转基因线虫将被用来区分暴露对不同神经元亚型、一般应激反应和蛋白酶体抑制的系统影响。还将使用全基因组寡核苷酸微阵列来分析响应细菌暴露的基因表达的差异变化，以确定受环境毒素调控的潜在基因。与公共卫生的相关性：遗传易感性和对环境侮辱的易感性之间的相互作用是帕金森病的核心。风险因素最好使用严格控制环境条件和遗传差异的系统进行评估。线虫是一种微小的蠕虫，恰好有8个多巴胺神经元，它与人类分享了大约一半的基因，是一种理想的系统，可以快速检测可能影响帕金森病发展的潜在环境毒素来源。