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

加州大学洛杉矶分校CGEP的目标是调查这样一种假设，即细胞的作用机制识别出 对于假定的环境毒物(PETS)，会显著增加帕金森病的风险；本项目将 重点开展啮齿动物调查。我们的团队确定了特定的细胞机制，这些机制会影响 PETS：蛋白酶体、微管完整性和乙醛脱氢酶脱毒。与.一起 影响多巴胺平衡的VMAT功能/表达改变，这些途径-可能影响 DA神经元对神经退行性变的易感性。我们建议将我们的互补专业知识用于 分子、神经化学、电生理和病毒基因传递方法来确定 在啮齿动物体内，这些细胞机制受到宠物的影响，并确定了额外的分子 以及由宠物引起的功能改变，这可能导致DA神经元的脆弱性增加。 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PETS是否 改变组织中K48和K63的泛化水平并增加线粒体醛的水平 多巴胺神经元中的脱氢酶底物。我们将发现新的基因参与细胞机制。 PETS对分离的黑质纹状体DA神经元转录组的影响 通过激光捕获显微切割，并用切片电生理学来鉴定其作用机制 PETS对DA神经元的细胞特性的研究。最后，我们将评估DA监管的作用 通过检测病毒或药物诱导的影响来调节PET毒性的细胞质水平 宠物对大鼠VMAT的影响。这些活体动物实验将在 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)增加人类患帕金森病的风险。反过来，转录组和电生理学分析在 项目3可能指出在果蝇遗传学研究中要研究的新的细胞途径(项目2) 在人类身上(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).