Role of Oxidative Stress and Protein Transporters in Glyphosate and Mancozeb Neur

描述（由申请人提供）：根据NIEHS的使命，了解环境如何影响人类疾病的病因和进展，本项目将研究两种高使用量农药对一般神经毒性和帕金森病（PD）的贡献。许多报告表明PD与农业环境和/或农药使用之间存在联系。其他数据表明，接触锰（Mn）也可能与疾病发病有关。有趣的是，许多农用化学品，以及锰，损害线粒体呼吸，导致细胞生存所需的能量生产不足。此外，线粒体功能障碍存在于PD患者的各种细胞群中。该项目的长期目标是了解农药可能导致PD相关脑区细胞死亡的机制。使用秀丽隐杆线虫（C. elegans），将测试以下假设：暴露于流行的农药代森锰锌（MZ）和触地（TD）通过线粒体抑制和/或增加氧化应激导致神经元变性。此外，假设这些农药可能通过特定的神经递质转运体摄取而进入脆弱的神经元。这些假设将通过三个具体目标进行测试：(1)确定哪些线粒体复合物受到抑制；(2)发现暴露于MZ和/或TD后特异性活性氧（ROS）增加；(3)确定MZ和TD是否通过单个神经递质或金属转运体进入神经元。为了监测神经退行性变，线虫的神经元群被绿色荧光蛋白（GFP）标记，将暴露于不同浓度的MZ或TD。随后将用各种化学试剂进行治疗，以确定线粒体功能是否受损。随后的研究将涉及用特定的神经递质转运拮抗剂预处理蠕虫，以确定是否评估在随后暴露于MZ或TD后变性是否减少。这些新研究的结果将进一步加深我们对农药暴露相关的神经毒性效应的理解。此外，这项工作将提供与含草甘膦除草剂（即TD， RoundUP）或含锰杀菌剂（即MZ， maneb）的细胞内运输机制相关的证据。预计这些数据不仅将进一步加深我们对一般农药神经毒性的理解，而且还将进一步了解环境毒物可能导致特发性PD病因的机制。

DESCRIPTION (provided by applicant): In accordance with the mission of the NIEHS, understanding how environment influences human disease etiology and progression, this project will examine the contribution of two high-usage pesticides to general neurotoxicity as well as Parkinson's disease (PD). Numerous reports suggest a link between PD and agricultural environments and/or pesticide usage. Other data indicate that exposure to manganese (Mn) also may be linked to disease onset. Interestingly, many agrochemicals, as well as Mn, impair mitochondrial respiration, resulting in an insufficient quantity of energy production required for cells survival. Furthermore, mitochondrial dysfunction is present in various cell populations in PD patients. The long-term goal of this project is to understand the mechanisms through which pesticides may contribute to cell death in brain regions associate with to PD. Using the nematode Caenorhabditis elegans (C. elegans), the following hypothesis will be tested: Exposure to the popular pesticides Mancozeb (MZ) and TouchDown (TD) leads to neuronal degeneration via mitochondrial inhibition and/or increased oxidative stress. Additionally, it is hypothesized that these pesticides may enter vulnerable neuron through uptake by specific neurotransmitter transporters. These hypotheses will be tested through three specific aims: (1) to establish which mitochondrial complexes are inhibited; (2) to discover the specific reactive oxygen species (ROS) increased following exposure to MZ and/or TD; and (3) to determine if MZ and TD enter neurons through individual neurotransmitter or metal transporters. In order to monitor neurodegeneration, C. elegans with neuronal populations tagged with green fluorescent protein (GFP) will be exposed to varying concentrations of MZ or TD. This will be followed by treatment with various chemical reporters to determine whether mitochondrial function is compromised. Subsequent studies will involve pretreatment of worms with specific neurotransmitter transporter antagonists to determine if assess whether degeneration is decreased following subsequent exposure to MZ or TD. Results from these novel studies will further our understanding of neurotoxic effects related to pesticide exposure. Additionally, this work will provide evidence related to mechanisms involved in intracellular transport of glyphosate-containing herbicides (i.e., TD, RoundUP), or manganese-containing fungicides (i.e., MZ, maneb). It is anticipated that these data will further our understanding not only of general pesticide neurotoxicity, but also of mechanisms through which environmental toxicants may contribute the etiology of idiopathic PD. PUBLIC HEALTH RELEVANCE: These studies are particularly timely and relevant to public health because of the link between Parkinson's disease (PD), the second most-common neurodegenerative disease in the United States, and pesticide exposure. Use of glyphosate-containing herbicides (i.e., RoundUp and TouchDown) is increasing exponentially in the general public and the agricultural community, while Mancozeb, a manganese-containing fungicide, is gaining increased market shares. These pesticides are suspected mitochondrial inhibitors, an observation commonly documented in various cell populations in PD patients.