Project 4: Biotransformation Gene-Environment Interactions in Coho Salmon

太平洋鲑鱼的数量在美国西部明显下降。特别令人关注的 在接触某些杀虫剂和微量金属的鲑鱼中发生的亚致死性神经损伤。这些 行为影响包括捕食者检测和猎物选择的损失，生殖时间的改变和损失 归巢。上述在个体中观察到的神经行为效应现在与 人口影响。鲑鱼嗅觉系统是环境污染物神经毒性的敏感靶点 化学品，包括金属和杀虫剂通常发现在超级基金网站。然而， 关于鱼类化学嗅觉神经毒性的机制。我们第一个资助周期的研究 产生了重要的发现，将在我们的竞争性更新中详细探讨。具体来说，我们知道 1）鲑鱼的嗅觉组织是化学生物转化的重要场所，并且特别地， 细胞色素P4503 A和黄素单加氧酶（FMO）似乎介导组织和化合物特异性 有机磷生物转化的差异对神经毒性有潜在影响，2）嗅觉 一种超级基金有机磷化学品（毒死蜱）和金属（铜）造成的伤害包括破坏 嗅觉信号传导通路的一个重要组成部分然而，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，而毒死蜱激活基因参与 嗅觉信号传导抑制; 3）转录特征可以帮助我们识别独特的基因 与混合物有关的目标，以及区分金属和有机磷酸盐驱动的影响。基于 我们的研究发现，竞争更新的目标是：1）利用cDNA克隆，重组蛋白， 表达，微阵列分析和酶的方法，以确定嗅觉CYP 3A的作用， 鲑鱼从淡水到淡水的迁移过程中有机磷酸盐神经毒性中的黄素单加氧酶 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 鉴定和区分铜和毒死蜱的重要嗅觉蛋白靶标的方法，4）使用 从上述研究中产生的一套嗅觉生物标志物，用于评估亚致死嗅觉 通过超级基金地点迁移的鲑鱼的神经毒性。

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.