Project 2: Pesticide Mechanisms and PD: Genetic Studies In Flies

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 对于推定的环境毒物(PETS)，会显著增加帕金森病的风险；该项目 (项目2)将集中在果蝇身上进行调查，以研究PET的作用机制及其 在相同的细胞通路中与遗传损伤相互作用。来自我们的流行病学和体外数据。 研究表明，暴露在宠物中会增加患帕金森病的风险，并提出了几种潜在的风险 宠物可能在多巴胺能神经元中产生毒素效应的机制。(DA)神经元：蛋白天冬氨酸， 微管功能和乙醛脱氢酶(ALDH)的解毒作用：我们还表明 泡状吗啡转运体(VMAT)的表达改变会影响DA神经元的易感性。 神经退行性变。在项目2上合作的三个果蝇实验室拥有丰富的使用经验 果蝇遗传学模拟神经退行性疾病和环境侮辱的贡献。 在这里，我们建议利用果蝇遗传学来研究：1)已知的果蝇的哪些生化活性 宠物导致DA细胞死亡，2)宠物和遗传损伤是否在同一生化途径中 可以增加DA细胞的死亡，以及3)VMAT的操作如何影响 宠物。抑制这些过程的药物和毒素具有多效性。我们建议准确地 使用分子遗传模拟来定义每个途径对抑制蛋白酶体的作用， 微管功能和乙醛脱氢酶。我们将使用的分子遗传试剂是 已经可获得或容易制造，并将包括RNA干扰以击倒泛素的表达 活化酶(E1)，两个典型的显性负性突变的错误表达。 蛋白酶体亚单位，在人类tau最长亚型的DA神经元中的表达，以及功能丧失 果蝇ALDH基因突变。项目2和项目1的成果将用于帮助发展啮齿动物 项目3和^中的模型有助于确定在人类遗传过程中要强调的生化途径 项目4的研究。PET暴露的Fly模型也将使我们能够在未来的目标中评估潜力 神经保护策略。

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 (Project 2) willfocus on investigations in Drosophila to Investigate the mechanisms of PET action and their interaction with .genetic lesions in the same cellular pathways. Epidemiological and in vitro data from our. group-have shown that exposure to PETs increases the risk of PD and suggested several potential mechanisms by which PETs may exert toxjc effects in dopaminergic.(DA) neurons: the proteaspme, microtubule function, and detoxification by aldehyde dehydrogenase (ALDH): We also have shown that altered expression of the vesicular mohoamine transporter (VMAT) affects the vulnerability of DA neurons to . neurodegeneration. The three Drosophila labs collaborating on Project 2 have extensive experience using Drosophila genetics to model neurodegenerative disorders and the contribution of environmental insults. Here, we propose to use Drosophila genetics to investigate: 1) which of the known biochemical activities of the PETs contribute to DA cell death, 2) whether PETs and genetic lesions in the same biochemical pathway can combine to increase DA cell death, and 3) how manipulation of VMAT affects the neurotoxicity of the PETs. Drugs and toxins that inhibit these processes have pleiotropic effects, and.we propose to precisely define the contribution of each pathway using molecular genetic mimics to inhibit the proteasome, microtubule function, and aldehyde dehydrogenase. The molecular genetic reagents we will use are either already available or readily made and will include RNA interference to knockdown expression of the ubiquitin activating enzyme (E1), misexpression of two well characterized dominant negative mutations.in 20S proteasome subunits, expression in DA neurons of the longest isoform of human tau, and loss of function mutations in Drosophila ALDH. The results of Project 2 and Project 1 will be used to help develop rodent models in Project 3, and^help determine biochemical pathways to be emphasized in the human genetic studies of Project 4. Fly models for PET exposure also will enable us in future aims to evaluate potential neuroprotective strategies.