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

UCLA-CGEP的目的是研究细胞作用机制的假设， 推定的环境毒物（PET）导致PD风险显著增加;本项目将 重点研究啮齿类动物。我们的小组确定了特定的细胞机制， PET：蛋白酶体、微管完整性和醛脱氢酶解毒。连同 改变VMAT功能/表达，影响多巴胺平衡，这些途径-可能会影响 DA神经元对神经变性的脆弱性。我们建议利用我们的互补专长， 分子、神经化学、电生理和病毒基因传递方法，以确定是否 这些细胞机制在啮齿动物脑中受体内PET的影响， 以及由PET诱导的功能改变，其可有助于增加DA神经元的脆弱性。 使用PET治疗方案，导致DA神经元的轻度功能障碍，我们将确定PET是否 改变组织中K48和K63泛素化水平，增加线粒体醛水平 脱氢酶底物。我们将确定参与细胞机制的新基因， 通过评估PETs诱导的黑质纹状体DA神经元分离的转录组变化来评估PETs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 多巴胺能神经元细胞特性的PET。最后，我们将评估DA监管的作用 细胞质水平在调节PET毒性中的作用 用PET处理的大鼠中VMAT的改变。这些体内动物实验将提供 在项目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，转录组和电生理分析， 项目3可能指出果蝇遗传学研究中要研究的新的细胞途径（项目2） 在人类（项目4）。

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).