Project 3: Pesticide Mechanisms and PD: In Vivo Studies In Rodents

项目 3：农药机制和 PD：啮齿类动物体内研究

• 批准号: 8292136
• 负责人: MARIE-FRANCOISE S CHESSELET
• 金额: $ 22.71万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8292136
• 关键词: Affect; Agriculture; Animal Experiments; Brain; Dopamine; Drosophila genus; Drug Metabolic Detoxication; Electrophysiology (science); Environment; Epidemiologic Studies; Equilibrium; Functional disorder; Gene Expression Profile; Genes; Genetic; Goals; Human; Investigation; Lead; Link; Microtubules; Mitochondria; Molecular; Nerve Degeneration; Parkinson Disease; Pathogenesis; Pathway interactions; Pesticides; Property; Rattus; Regulation; Risk; Rodent; Role; Slice; Tissues; Toxic Environmental Substances; Toxic effect; Treatment Protocols; Ubiquitination; aldehyde dehydrogenases; base; cohort; dopaminergic neuron; environmental agent; in vivo; laser capture microdissection; multicatalytic endopeptidase complex; neurochemistry; novel; pesticide exposure; viral gene delivery

The goal of the UCLA-CGEP is to investigate the hypothesis that the cellular mechanisms of action identified for Putative Environmental Toxicants (PETs) contribute to a significant increase in PD risk; this project will focus on investigations in rodents. Our group identified specific cellular mechanisms that are affected by PETs: the proteasome, microtubule integrity, and aldehyde dehydrogenase detoxification. Together with altered VMAT function/expression that influences dopamine balance, these pathways-may affect the vulnerability of DA neurons to neurodegeneration. We propose to use our complementary expertise with molecular, neurochemical, electrophysiological, and viral gene delivery approaches to determine whether these cellular mechanisms are affected by PETs in vivo in rodent brains and identify additional molecular and functional alterations induced by PETs that can contribute to increased vulnerability of DA neurons. Using PET treatment regimens that lead to mild dysfunction of DA neurons we will determine whether PETs alter levels of K48 and K63 ubiquination in tissue and increase levels of mitochondrial aldehyde dehydrogenase substrates in DA neurons. We will identify new genes involved in the cellular mechanism of actions of PETs by assessing transcriptome changes induced by PETs in nigrostriatal DA neurons isolated by laser capture microdissection, and use slice electrophysiology to identify the mechanism of action of PETs on the cellular properties of DA neurons. Finally, we will assess the role of regulation of DA cytoplasmic levels in modulating PET toxicity by examining the effects of virally or pharmacologically induced alterations of VMAT in rats treated with PETs. These in vivo animal experiments will provide a link between molecular mechanisms identified in projects 1 and 2 and the toxicity of compounds found by our group (Project 4) to increase PD risk in humans. In turn, the transcriptome and electropysiological analyses in Project 3 may point to new cellular pathways to be investigated in genetic studies in Drosophila (Project 2) and in humans (Project 4).

UCLA-CGEP 的目标是研究以下假设：已确定的细胞作用机制 假定环境毒物 (PET) 会导致 PD 风险显着增加；该项目将 重点关注啮齿类动物的研究。我们的小组确定了受以下因素影响的特定细胞机制 PET：蛋白酶体、微管完整性和乙醛脱氢酶解毒。连同 改变影响多巴胺平衡的 VMAT 功能/表达，这些途径可能会影响 DA 神经元对神经退行性变的脆弱性。我们建议利用我们的互补专业知识 分子、神经化学、电生理学和病毒基因传递方法，以确定是否 这些细胞机制在啮齿类动物大脑中受到体内 PET 的影响，并识别出其他分子 PET 引起的功能改变可能会增加 DA 神经元的脆弱性。 使用导致 DA 神经元轻度功能障碍的 PET 治疗方案，我们将确定 PET 是否 改变组织中 K48 和 K63 泛素化水平并增加线粒体醛水平 DA 神经元中的脱氢酶底物。我们将鉴定参与细胞机制的新基因 通过评估 PET 在分离的黑质纹状体 DA 神经元中诱导的转录组变化来评估 PET 的作用 通过激光捕获显微切割，并使用切片电生理学来确定作用机制 PET 对 DA 神经元细胞特性的影响。最后，我们将评估 DA 监管的作用 通过检查病毒或药物诱导的影响来调节 PET 毒性的细胞质水平 用 PET 治疗的大鼠 VMAT 的变化。这些体内动物实验将提供之间的联系 项目1和2中确定的分子机制以及我们小组发现的化合物的毒性 （项目 4）增加人类 PD 风险。反过来，转录组和电生理学分析 项目 3 可能指出果蝇遗传研究中待研究的新细胞途径（项目 2） 以及人类（项目 4）。