Mechanism of stress induced developmental abnormalities

应激诱发发育异常的机制

• 批准号: 6442559
• 负责人: JONATHAN H FREEDMAN
• 金额: $ 8.64万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6442559
• 关键词: AP1 protein; biological signal transduction; cadmium; cell line; chlorohydrocarbon insecticide; copper; environmental toxicology; free radical oxygen; gene expression; genetic transcription; genetically modified animals; growth /development; hazardous substances; insecticide biological effect; insecticides; metallothionein; oxidative stress; protooncogene; regulatory gene; reporter genes; tissue /cell culture; transfection; zebrafish

Exposure to metals (cadmium, cooper) and organic pesticides (dieldrin, chlorpyrifos) disrupts normal cellular development and differentiation. These hazardous chemicals have been shown to induce intracellular oxidative stress and produce reactive oxygen species (ROS). ROS in turn can stimulate activating protein-1 (AP-1) binding, as well as increase c- jun and c-fos protein and mRNA levels. It has been shown that ROS increases in AP-1 binding activating c-jun N-terminal kinase (JNK) or extracellularly responsive kinases (ERK). Ultimately, the generation of intracellular oxidative stress can disrupt normal development by "inappropriately" activating these signal transduction cascades to induce the transcription of genes that affect cell growth and differentiation. The hypothesis that cadmium, copper, dieldrin and chlorpyrifos increase the intracellular levels of ROS to affect c-jun levels, and increase AP-1 binding activity will be investigated. To explore this hypothesis two model system two model systems will be used: cultured mammalian cells and zebrafish. The studies proposed using cultured cell lines will investigate molecular and cellular aspects of hazardous chemical exposure on development. As a physiologic end-point, the effects of cadmium and copper exposure on zebrafish development will be examined. There are four specific aims in this project: (1) Characterize the effects of metals and pesticides on metallothionein and proto-oncogene expression. (2) Identify upstream regulatory elements that mediate chemically induced transcription. (3) Determine the effects of hazardous chemical exposure on the activities of (a) AP-1 and (b) signal transduction cascades that regulate Ap-1 pathways that regulate development in zebrafish. The information and concepts obtained from the proposed studies directly address several of the goals outlined in the Superfund Research Program, which is to understand the human health and environmental effects associated with hazardous waste. Specifically, the research outlined in this project will address the mechanistic basis for cellular and molecular perturbations and associated health effects that are due to exposure to hazardous substances.

接触金属（镉、铜）和有机农药（狄氏剂、毒死蜱）会破坏正常的细胞发育和分化。这些有害化学物质已被证明可诱导细胞内氧化应激并产生活性氧（ROS）。ROS进而刺激活化蛋白-1 （AP-1）结合，增加c- jun和c-fos蛋白及mRNA水平。研究表明，活性氧增加AP-1结合激活c-jun n -末端激酶（JNK）或细胞外反应激酶（ERK）。最终，细胞内氧化应激的产生可以通过“不恰当地”激活这些信号转导级联来诱导影响细胞生长和分化的基因转录，从而破坏正常发育。假设镉、铜、狄氏剂和毒死蜱增加细胞内ROS水平影响c-jun水平，并增加AP-1结合活性。为了探索这一假设，我们将使用两个模型系统：培养的哺乳动物细胞和斑马鱼。建议使用培养细胞系的研究将研究危险化学品暴露对发育的分子和细胞方面的影响。作为一个生理终点，镉和铜暴露对斑马鱼发育的影响将被检查。本项目有四个具体目标：(1)研究金属和农药对金属硫蛋白和原癌基因表达的影响。(2)鉴定介导化学诱导转录的上游调控元件。(3)确定危险化学品暴露对(a) AP-1和(b)调节斑马鱼发育的AP-1通路的信号转导级联活性的影响。从拟议的研究中获得的信息和概念直接涉及超级基金研究方案中概述的几个目标，即了解与危险废物有关的人类健康和环境影响。具体而言，本项目概述的研究将探讨由于接触有害物质而引起的细胞和分子扰动以及相关的健康影响的机制基础。