Zebrafish as a Detector of Organophosphate Exposure

斑马鱼作为有机磷暴露检测器

• 批准号: 6900497
• 负责人: Elwood Albert Linney
• 金额: $ 22.87万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6900497
• 关键词: acetylcholinesterase; animal developmental psychology; behavioral /social science research tag; biosensor device; developmental disease /disorder; embryo /fetus toxicology; environmental exposure; environmental toxicology; esterase inhibitor; gene expression; genetically modified animals; killifish; learning; microarray technology; morphology; nonmammalian vertebrate embryology; organophosphorus insecticide; zebrafish

Our working hypothesis is that there is a hierarchy of effects of organophosphate embryonic organophosphate exposure but selective targets and developmental windows are responsible for adult effects. We plan to use zebrafish to screen organophosphate pesticides to examine embryonic and larval effects upon exposure and in collaboration with the Levin laboratory, to investigate and compare effects of these exposures upon learning. Our intention is to screen at least 3 additional esterase inhibitors (along with chlorpyrifos: parathion, diazinon from the superfund list, and the non-superfund list but commonly used malathion). The four specific aims are: 1) to perform dosage studies with selected transgenic lines for the first 5 days post-fertilization and to score for a)inhibition of acetylcholine esterase and b)morphological effects?preliminary behavioral differences will be noted for potential analysis of adults by E. Levin 2) to perform 5 day time courses with selected exposures and to perform microarray analyses to identify genes, clusters of genes and time course of gene expression effects 3) through analysis of expression work to a) identify potential unique genes(per chemical) that are markedly up-regulated(as candidate genes for isolating their promoters to create inhibitor-specific transgenic biosensor) and b) to identify potential genes and their time-course that are positively or negatively affected by the exposure to differentiate effects of different inhibitors and potentially identify pathways affected?this could include in situ localization of the genes affected by the exposure 4) from specific aim 3) the generation of organophosphate- specific biosensor transgenic fish based upon unique genes up-regulated by each inhibitor, evaluate transgenics with inhibitor exposures and determine whether battery of transgenic fish can be used to assay mixtures of inhibitors.

我们的工作假设是，有机磷酸盐对胚胎的影响存在层次结构。 有机磷暴露但选择性目标和发育窗口是造成成人影响的原因。我们计划使用斑马鱼筛选有机磷农药，以检查接触后对胚胎和幼虫的影响，并与莱文实验室合作，调查和比较这些接触对学习的影响。我们的目的是筛选至少 3 种额外的酯酶抑制剂（以及毒死蜱：超级基金清单中的对硫磷、二嗪农，以及非超级基金清单但常用的马拉硫磷）。四个具体目标是：1) 在受精后的前 5 天对选定的转基因品系进行剂量研究，并对 a) 乙酰胆碱酯酶的抑制和 b) 形态效应进行评分？ E. Levin 将注意到初步行为差异，以便对成虫进行潜在分析 2) 在选定的暴露下进行 5 天的课程，并进行微阵列分析以识别基因、基因簇和 基因表达效应的时间进程 3) 通过表达工作分析，a) 识别显着上调的潜在独特基因（每种化学物质）（作为分离其启动子以创建抑制剂特异性转基因生物传感器的候选基因），b) 识别受到不同抑制剂不同效应的正面或负面影响的潜在基因及其时间进程，并可能识别受影响的途径？这可能包括受不同抑制剂影响的基因的原位定位 暴露4)来自特定目标3)基于每种抑制剂上调的独特基因生成有机磷酸盐特异性生物传感器转基因鱼，评估抑制剂暴露的转基因并确定是否可以使用转基因鱼组来测定抑制剂混合物。