Mechanisms of Embryo Response to Oxidative Stress

胚胎对氧化应激的反应机制

基本信息

批准号：
8051862
负责人：
Mark E Hahn
金额：
$ 38.07万
依托单位：
WOODS HOLE OCEANOGRAPHIC INSTITUTION
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-06-15 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8051862
关键词：
Affect Animals Antioxidants Basic Science Binding Biological Models Cardiovascular Diseases Chemicals Cysteine Development Diquat Disease Drug or chemical Tissue Distribution Embryo Embryonic Development Environmental Exposure Family Fishes Foundations Gene Expression Generations Genes Genetic Transcription Genomics Glutamate-Cysteine Ligase Glutathione Glutathione S-Transferase Growth Health Homologous Gene Human Individual Life Ligase Link Mammals Mediating Modeling Molecular NAD(P)H dehydrogenase (quinone) 1, human NF-E2-related factor 2 NQO1 gene Neurodegenerative Disorders Oligonucleotides Oxidants Oxidative Stress Phenotype Predisposition Production Protein Biosynthesis Protein Family Protein Isoforms Proteins Reactive Oxygen Species Regulation Regulatory Element Reporter Genes Research Response Elements Reverse Transcriptase Polymerase Chain Reaction Risk Assessment Role Screening procedure Staging Sulforaphane Superoxide Dismutase Testing Time Toxic effect Transgenes Transgenic Organisms Zebrafish bZIP Domain biological adaptation to stress experimental analysis human disease in vivo insight knock-down mature animal member paralogous gene promoter public health relevance research study response tool toxicant

Affect Animals Antioxidants Basic Science Binding Biological Models Cardiovascular Diseases Chemicals Cysteine Development Diquat Disease Drug or chemical Tissue Distribution Embryo Embryonic Development Environmental Exposure Family Fishes Foundations Gene Expression Generations Genes Genetic Transcription Genomics Glutamate-Cysteine Ligase Glutathione Glutathione S-Transferase Growth Health Homologous Gene Human Individual Life Ligase Link Mammals Mediating Modeling Molecular NAD(P)H dehydrogenase (quinone) 1, human NF-E2-related factor 2 NQO1 gene Neurodegenerative Disorders Oligonucleotides Oxidants Oxidative Stress Phenotype Predisposition Production Protein Biosynthesis Protein Family Protein Isoforms Proteins Reactive Oxygen Species Regulation Regulatory Element Reporter Genes Research Response Elements Reverse Transcriptase Polymerase Chain Reaction Risk Assessment Role Screening procedure Staging Sulforaphane Superoxide Dismutase Testing Time Toxic effect Transgenes Transgenic Organisms Zebrafish bZIP Domain biological adaptation to stress experimental analysis human disease in vivo insight knock-down mature animal member paralogous gene promoter public health relevance research study response tool toxicant

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项目摘要

DESCRIPTION (provided by applicant): Oxidative stress resulting from environmental exposures is associated with a variety of human diseases ranging from chemical teratogenesis to cardiovascular and neurodegenerative diseases. Developing animals appear to be especially sensitive to chemicals causing oxidative stress. The expression and inducibility of anti-oxidant defenses are critical factors affecting susceptibility to oxidants at these early life stages, but the ontogenic development of these responses in embryos is not well understood. In adult animals, oxidants initiate an anti-oxidant response by activating NF-E2-related factor 2 (NRF2) and related proteins, which bind to the anti-oxidant response element and activate transcription of genes such as glutathione S-transferases, NAD(P)H-quinone oxidoreductase, glutamyl-cysteine ligase, and superoxide dismutase. The overall objective of the research proposed here is to elucidate the mechanisms by which vertebrate embryos respond to oxidative stress during development. We will test the central hypothesis that responsiveness to oxidative stress and the set of regulated genes vary during development. Because of these developmental differences, some stages may be more sensitive to oxidative stress-induced damage. These studies will be performed in vivo using embryos of the zebrafish (Danio rerio), a valuable model in which to examine mechanisms of toxicity in developing animals and to screen chemicals for developmental toxicity. Aim 1 will use transcriptional profiling and phenotypic anchoring to identify the core set of genes that comprise the oxidative stress response in embryos, establish how the responsiveness and composition of the core set of oxidant-responsive genes vary with developmental stage of embryos, and determine how the timing and gene profile of the oxidative stress response differ in embryos exposed to structurally and mechanistically distinct activators of NRF2 (tBHQ, diquat, sulforaphane). Aim 2 will elucidate the roles of different NRF paralogs in the transcriptional response to oxidative stress during development in embryos in vivo, using targeted knock-down of NRF protein synthesis with morpholino oligonucleotides. Aim 3 will establish the mechanism of regulation of anti-oxidant response genes during embryonic development through computational and in vivo experimental analysis of oxidant-responsive gene promoters, leading to the generation of a stable line of transgenic zebrafish expressing a reporter gene (GFP) in response to oxidative stress. Finally, we will test a set of mammalian developmental toxicants for the ability to activate the transgene in embryos. The results of these studies will establish the composition and ontogeny of the transcriptional response to oxidative stress in vertebrate embryos, elucidate fundamental mechanisms underlying this response, generate tools for screening chemicals for activity as developmental toxicants or antioxidants, and provide insight into the role of oxidative stress in human disease. PUBLIC HEALTH RELEVANCE: Oxidative stress is involved in a variety of environmentally influenced human diseases ranging from chemical teratogenesis to cardiovascular and neurodegenerative diseases. The proposed research will enhance our basic understanding of how some chemicals interfere with embryonic development by generating oxidative stress and how embryos can mitigate these effects. This research will i) determine the basal expression and inducibility of anti-oxidant defenses during embryonic development, ii) elucidate fundamental mechanisms of the response of vertebrate embryos to oxidative stress, iii) establish a model system for screening chemicals for activity as developmental toxicants or antioxidants, and iv) provide a mechanistic foundation that will facilitate the extrapolation of results obtained in zebrafish to humans, supporting risk assessment and providing insight into the role of oxidative stress in human disease.

描述（由申请人提供）：环境暴露引起的氧化应激与各种人类疾病有关，范围从化学畸胎发生到心血管和神经退行性疾病。发育中的动物似乎对引起氧化应激的化学物质特别敏感。抗氧化剂防御措施的表达和诱导性是影响这些早期生命阶段氧化剂易感性的关键因素，但是对胚胎中这些反应的个体发育尚不清楚。在成年动物中，氧化剂通过激活NF-E2相关因子2（NRF2）和相关蛋白质引发抗氧化剂反应，该因子与抗氧化剂反应元件结合并激活基因的转录，例如谷胱甘肽S-转移酶，NAD（P）H- Quinone氧化氧化物酶酶酶，粘液氨基氨基酶ligasase ligasase，and ligase ligasase stase stase stase stase stase＆procrets ult and suprart and ult and and suprart and ult and。此处提出的研究的总体目的是阐明脊椎动物胚胎在发育过程中对氧化应激反应的机制。我们将检验一个中心假设，即对氧化应激的反应性和在发育过程中的受调节基因集有所不同。由于这些发育差异，某些阶段可能对氧化应激引起的损伤更敏感。这些研究将使用斑马鱼（Danio Rerio）的胚胎在体内进行，这是一个有价值的模型，其中旨在检查发育中动物中毒性的机制，并筛选出化学物质的发育毒性。 AIM 1将使用转录分析和表型锚定来识别构成胚胎中氧化应激反应的核心基因集，并确定氧化剂反应基因的核心核心集合和组成如何随胚胎的发育阶段而变化，并确定在嵌入型氧化应激术中的敏感性和基因在结构上的敏感性和基因的结构敏感性和基因的结构性响应方式以及结构性的机制差异。（TBHQ，Diquat，Sulforaphane）。 AIM 2使用靶向敲击NRF蛋白质合成与morpholino寡核苷酸的靶向敲击，将阐明不同NRF旁系同源物在体内发育过程中对氧化应激的转录反应中的作用。 AIM 3将通过计算和体内实验分析对氧化剂反应性基因启动子进行调节抗氧化反应基因的调节机制，从而导致产生稳定的转基因斑马鱼，表达对氧化应激的响应，表达报告基因的基因（GFP）。最后，我们将测试一组哺乳动物发育毒物，以便激活胚胎中的转基因的能力。这些研究的结果将建立脊椎动物胚胎中对氧化应激的转录反应的组成和个体，阐明了这种反应的基本机制，生成了用于筛选化学物质作为发育毒理或抗氧化剂的化学物质的工具，并为氧化应激在人类疾病中的作用提供了洞察力。公共卫生相关性：氧化应激与各种受环境影响的人类疾病有关，从化学致畸作用到心血管和神经退行性疾病。拟议的研究将增强我们对某些化学物质如何通过产生氧化应激以及胚胎如何减轻这些作用来干扰胚胎发育的基本理解。这项研究将i）确定在胚胎发育过程中抗氧化剂防御的基础表达和诱导性，ii）阐明脊椎动物胚胎对氧化应激的反应的基本机制，iii）建立一种筛选化学物质的模型系统，用于将化学物质作为发育毒性或抗氧化剂的发展，并为统一的基础提供了范围，以使其对统一的基础，以使范围内的基础构成范围的范围，并构成了范围内的范围。风险评估并提供有关氧化应激在人类疾病中的作用的见解。