Project 4: Biotransformation Gene-Environment Interactions in Coho Salmon

美国西部的太平洋鲑鱼数量明显减少。特别令人担忧的是 在暴露于某些杀虫剂和微量金属的鲑鱼中发生亚致死性神经损伤。这些 行为影响包括失去捕食者检测和猎物选择、繁殖时机改变和丧失 归宿之路。在个体中观察到的上述神经行为影响现在与 对人口的影响。鲑鱼嗅觉系统是环境神经毒性的敏感靶点。 化学品，包括超级基金网站上常见的金属和杀虫剂。然而，人们对此知之甚少 关于鱼的化学嗅觉神经毒性的机制。我们第一个资金周期的研究已经 产生了重要的发现，将在我们的竞争更新中详细探索。具体来说，我们知道 认为：1)鲑鱼的嗅觉组织是化学生物转化的重要部位，尤其是， 细胞色素P4503A和黄素单加氧酶(FMO)似乎介导组织和化合物的特异性 有机磷生物转化的差异及其对神经毒性的潜在影响，2)嗅觉 超级基金模型有机磷化学品(毒死蜱)和金属(铜)造成的伤害涉及破坏 嗅觉信号转导通路。然而，铜主要影响G蛋白偶联嗅觉 受体信号，可能是通过氧化应激，而毒死蜱激活参与 嗅觉信号转导的抑制，以及3)转录信号可以帮助我们识别独特的基因 与混合物相关的目标，以及区分金属和有机磷驱动的影响。基于 我们的发现，竞争更新的目标是：1)使用cdna克隆，重组蛋白 表达，微阵列分析和酶方法确定嗅觉细胞色素P3A和 黄素单加氧酶在鲑鱼从淡水到淡水迁移过程中的有机磷神经毒性 盐水，2)使用原位杂交和免疫组织化学分析结合行为研究 了解氧化应激在铜和镉介导的嗅觉损伤中的作用，3)使用蛋白质组学 识别和区分铜和毒死蜱的重要嗅觉蛋白靶标的方法，4)用途 从前述研究中产生的一套嗅觉生物标志物来评估亚致死性嗅觉 通过超级基金网站迁徙的鲑鱼的神经毒性。

Pacific salmon populations have declined markedly in the Western United States. Of particular concern has been sublethal neurological injury occurring in salmon exposed to certain pesticides and trace metals. These behavioral impacts include loss of predator detection and prey selection, altered reproductive timing and loss of homing. These aforementioned neurobehavioral effects observed in individuals are now linked to population impacts. The salmon olfactory system is a sensitive target for the neurotoxicity of environmental chemicals, including metals and pesticides commonly found in Superfund sites. However, little is known about the mechanisms of chemical olfactory neurotoxicity in fish. Studies from our first funding cycle have produced important findings that will be explored in detail in our competitive renewal. Specifically, we know that: 1) the olfactory tissues of salmon are important site of chemical biotransformation, and in particular, cytochrome P4503A and flavin monooxygenases (FMO) appear to mediate tissue- and compound-specific differences in organophosphate biotransformation with potential impacts on neurotoxicity, 2) the olfactory injury by a model superfund organophosphate chemical (chlorpyrifos) and metal (copper) involves disruption of olfactory signal transduction pathways. However, copper primarily impacts G-protein coupled olfactory receptor signaling, likely through oxidative stress, whereas chlorpyrifos activates genes involved in the inhibition of olfactory signal transduction, and 3) transcriptional signatures can help us identify unique gene targets relevant to mixtures, as well differentiating metal- and organophosphate-driven affects. Based upon our findings, the objectives of the competing renewal are to: 1) use cDNA cloning, recombinant protein expression, microarray analysis and enzymatic approaches to determine the role of olfactory CYP3A and flavin monooxygenases in organophosphate neurotoxicity in salmon during movement from freshwater to saltwater, 2) use in situ hybridization and immunohistochemistry analyses coupled with behavioral studies to understand the role of oxidative stress in copper and cadmium-mediated olfactory injury, 3) use proteomics approaches to identify and discriminate important olfactory protein targets of copper and chlorpyrifos, 4) use a suite of olfactory biomarkers generated from the aforementioned studies to assess sublethal olfactory neurotoxicity in salmon migrating through Superfund sites.