Investigating Genetic and Environmental Etiologies of Parkinson's Ds in Zebrafish

研究斑马鱼帕金森病的遗传和环境病因

基本信息

批准号：
7659344
负责人：
JEFF Michael BRONSTEIN
金额：
$ 23.1万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-03-06 至 2011-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7659344
关键词：
Adult Affect American Animal Model Animals Apomorphine Automobile Driving Behavior Behavioral Benomyl Brain Caenorhabditis elegans Cell Death Cell Line Cell physiology Cells Characteristics Data Detection Development Disease Dopamine Dopaminergic Cell Drosophila genus DsRed Embryo Environment Environmental Risk Factor Epidemiologic Studies Etiology Evaluation Exhibits Exposure to Fishes Fluorescence Functional disorder Gene Mutation Genes Genetic Genetic screening method Goals Green Fluorescent Proteins Human Image Immunohistochemistry LRRK2 gene Larva Lasers Lesion Life Life Cycle Stages Measures Midbrain structure Modeling Modification Motor Neuroblastoma Parkinson Disease Pathogenesis Pathology Pesticides Phenotype Photons Proteasome Inhibition Proteasome Inhibitor Proteins Public Health Publishing Reporter Risk Risk Factors Role Rotenone Smell Perception Staining method Stains Swimming System Techniques Testing Toxic Environmental Substances Toxic effect Toxin Transgenes Transgenic Organisms Tyrosine 3-Monooxygenase Ubiquitin Vertebrates Zebrafish Ziram alpha synuclein behavior measurement disorder risk dopamine system epoxomicin exposed human population gene environment interaction high throughput screening in vivo inhibitor/antagonist molecular imaging multicatalytic endopeptidase complex novel olfactory bulb overexpression promoter protein expression public health relevance research study synuclein transgene expression

项目摘要

DESCRIPTION (provided by applicant): The cause of Parkinson's disease (PD) remains elusive but almost certainly involves genetic and environment factors. A number of potential risk factor genes and environmental toxins (especially pesticides) have been implicated but it is still not known if these factors actually cause PD. Current animal models to test causality of these factors and interactions between the two are inadequate. We propose to use zebrafish to test potential genetic and environmental causes of PD because they have several advantages over current animal models. Zebrafish are vertebrates, small, have a short life cycle, are relatively easy to insert transgenes, and the larvae are transparent enabling imaging of molecular and cellular processes in living animals. Behavior can also be readily measured. We propose to take advantage of these unique characteristics to determine the potential etiological roles of PD risk factor genes and environmental toxins implicated in PD and interactions between genes and the environment. We hypothesize that some toxins will alter locomotor and olfactory behavior and induce selective dopaminergic cell death and pathology similar to that seen in PD. It is also possible that the toxicity of some genes and toxins will only be apparent in combination (gene-environment interaction). Specifically, we will characterize the behavioral effects resulting from lesions in specific dopaminergic cell clusters made with a 2-photon laser and identified using a zebrafish tyrosine hydroxylase promoter driving green fluorescent protein (zTH-GFP). We will then be able to use behavioral measurements to screen for effects of PD genes and toxins on the dopaminergic system. PD- associated genes to be tested include alpha-synuclein and LRRK2. Toxins to be tested will include the ubiquitin-proteasome system (UPS) inhibitor epoxomicin, and pesticides identified to be inhibitors of the UPS using a high throughput screen performed in a neuroblastoma cell line. Expression of the zTH-GFP transgene will also facilitate pathological evaluation of toxin-exposed zebrafish and fish expressing PD genes. These studies will not only provide valuable information on the contributions PD genes and environmental toxins make to the pathogenesis of PD, but also will create a powerful model to test potential disease modifying therapies. PUBLIC HEALTH RELEVANCE: Parkinson's disease (PD) affects approximately 1 million Americans but the cause remains elusive. We propose to investigate genetic and environmental contributions to the development of PD using a novel zebrafish model. The results of these experiments will provide valuable information into the cause of PD and move us closer to the development of meaningful disease modifying therapies.

描述（由申请人提供）：帕金森氏病（PD）的原因仍然难以捉摸，但几乎肯定涉及遗传和环境因素。已经涉及许多潜在的危险因素基因和环境毒素（尤其是农药），但仍不知道这些因素是否真的引起PD。当前的动物模型测试这些因素的因果关系以及两者之间的相互作用不足。我们建议使用斑马鱼测试PD的潜在遗传和环境原因，因为它们比当前动物模型具有多个优势。斑马鱼是脊椎动物，小，生命周期短，相对容易插入转基因，幼虫是透明的，可以使活体动物中分子和细胞过程的成像进行成像。行为也可以很容易地衡量。我们建议利用这些独特的特征来确定与PD有关的PD危险因素基因和环境毒素的潜在病因，以及基因与环境之间的相互作用。我们假设某些毒素会改变运动和嗅觉行为，并诱导选择性多巴胺能细胞死亡和病理学类似于PD中所见。某些基因和毒素的毒性也可能仅在组合（基因 - 环境相互作用）中很明显。具体而言，我们将表征由特定多巴胺能细胞簇在用2光子激光制成的特定多巴胺能细胞簇产生的行为作用，并使用斑马鱼酪氨酸羟化酶启动子驱动绿色荧光蛋白（ZTH-GFP）鉴定。然后，我们将能够使用行为测量值来筛选PD基因和毒素对多巴胺能系统的影响。要测试的PD相关基因包括α-核蛋白和LRRK2。要测试的毒素将包括泛素 - 蛋白酶体系统（UPS）抑制剂环氧素，以及使用在神经母细胞瘤细胞系中执行的高吞吐量筛查的鉴定为UPS抑制剂的农药。 ZTH-GFP转基因的表达还将促进毒素暴露的斑马鱼和表达PD基因的病理评估。这些研究不仅将提供有关PD基因和环境毒素对PD发病机理的贡献的宝贵信息，而且还将创建一个强大的模型来测试潜在的修饰疗法。公共卫生相关性：帕金森氏病（PD）影响了大约100万美国人，但原因仍然难以捉摸。我们建议使用新型斑马鱼模型研究对PD发展的遗传和环境贡献。这些实验的结果将为PD的原因提供宝贵的信息，并使我们更接近有意义的疾病修饰疗法的发展。