TCDD- INDUCED CARDIOTOXICITY IN DEVELOPING ZEBRAFISH

TCDD-在发育中的斑马鱼中诱导心脏毒性

基本信息

批准号：
8215401
负责人：
WARREN no middle name HEIDEMAN
金额：
$ 1.94万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-01-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8215401
关键词：
ARNT gene Address Adolescent Adult Affect Aryl Hydrocarbon Receptor Binding Binding Sites Biological Assay Biological Models Cardiotoxicity Cell Transplantation Cell physiology Cells Chemicals Chromatin Structure Complex Congenital Abnormality Congenital Heart Defects Coupled DNA Binding DNA Sequence Differentiation and Growth Dioxins Dominant-Negative Mutation Embryo Embryonic Heart Environmental Pollution Epidemiology Exposure to Gene Expression Gene Expression Regulation Gene Targeting Genes Goals Growth Health Heart Heart Atrium Heart failure Human Imaging Techniques Immigration Immunohistochemistry In Vitro Jaw Label Laboratories Laboratory Research Ligands Mediating Methods Modeling Molecular Muscle Myocardial Myocardium Natural regeneration Pattern Poison Polychlorinated Biphenyls Process RNA Receptor Activation Recruitment Activity Resistance Specificity Surgical wound System Testing Tetrachlorodibenzodioxin Tissues Toxic effect Transcription Coactivator Transgenic Organisms Transplantation Work Zebrafish base blastomere structure cardiogenesis cell motility cell type chromatin immunoprecipitation heart cell in vivo migration prevent promoter receptor binding research study response time use

ARNT gene Address Adolescent Adult Affect Aryl Hydrocarbon Receptor Binding Binding Sites Biological Assay Biological Models Cardiotoxicity Cell Transplantation Cell physiology Cells Chemicals Chromatin Structure Complex Congenital Abnormality Congenital Heart Defects Coupled DNA Binding DNA Sequence Differentiation and Growth Dioxins Dominant-Negative Mutation Embryo Embryonic Heart Environmental Pollution Epidemiology Exposure to Gene Expression Gene Expression Regulation Gene Targeting Genes Goals Growth Health Heart Heart Atrium Heart failure Human Imaging Techniques Immigration Immunohistochemistry In Vitro Jaw Label Laboratories Laboratory Research Ligands Mediating Methods Modeling Molecular Muscle Myocardial Myocardium Natural regeneration Pattern Poison Polychlorinated Biphenyls Process RNA Receptor Activation Recruitment Activity Resistance Specificity Surgical wound System Testing Tetrachlorodibenzodioxin Tissues Toxic effect Transcription Coactivator Transgenic Organisms Transplantation Work Zebrafish base blastomere structure cardiogenesis cell motility cell type chromatin immunoprecipitation heart cell in vivo migration prevent promoter receptor binding research study response time use

展开

项目摘要

DESCRIPTION (provided by applicant): Epidemiological and laboratory evidence indicates that environmental contaminants such as TCDD affect vertebrate heart development. The receptor for TCDD is the Aryl Hydrocarbon Receptor (AHR), a ligand-activated transcriptional activator. We have used zebrafish, an ideal system for studying heart development in vivo to document the cardiotoxic response to TCDD. These results support a model in which TCDD disrupts normal gene regulation in the developing heart. Our microarray experiments have defined the pattern of gene expression changes produced by TCDD in zebrafish embryonic and adult hearts. We find that AHR activation regulates a different set of genes in embryonic heart cells than in juvenile heart cells. Adult hearts are resistant to TCDD cardiotoxicity, except when regenerating lost tissue after surgical wounding. The sensitivity to TCDD in the embryonic and regenerating adult heart correlate with periods in which epicardial cell migration, division, and differentiation are critical to the heart. This suggests that TCDD may be affecting epicardial cell function. We have developed methods for transplantation of cells from embryonic zebrafish hearts into the wounded adult muscle. These embryonic cells become incorporated into the wounded heart muscle. This project is focused on the following questions: 7 Are the cardiotoxic effects of TCDD mediated by AHR activation in the heart? This will be addressed by using transgenic zebrafish to manipulate AHR activity specifically in the heart. 7 Does TCDD alter epicardial function in the embryonic and regenerating adult heart? This will be addressed by determining whether TCDD blocks epicardial cell formation and migration in the embryonic heart. In addition, we will determine whether TCDD alters the activation, migration, growth, and differentiation of embryonic-like epicardial cells in the adult heart in response to wounding. We will compare the genes affected by TCDD in adult wounded hearts to those affected by TCDD in embryonic hearts. Finally, we will determine whether TCDD prevents transplantation of embryonic heart cells into wounded zebrafish myocardium. 7 Why does TCDD activation of AHR affect a different set of genes in the embryonic heart than in the juvenile heart or the developing jaw? We will test the hypothesis that the AHR complex is different in different tissues, producing different DNA binding site specificity in different tissues. As an alternative, we will test the hypothesis that the chromatin structure at a given gene may allow AHR binding in the embryonic heart, but in other tissues the chromatin structure at this gene would be different, preventing AHR binding. This will be tested using ChIP assays coupled to microarrays (ChIP-chip assays). PUBLIC HEALTH RELEVANCE: Heart defects are among the most common birth defects in humans, and recent epidemiological and laboratory research suggest that heart defects may be caused by exposure to chemicals such as 2,3,7,8-tetrachlorodibenzo-p-dioxin, and polychlorinated biphenyls, commonly known as dioxin and PCBs respectively. This project will be based on the zebrafish, Danio rerio, an important non-human model system widely used for studying heart development. The work will identify mechanisms by which toxic compounds disrupt heart development, and explores the possibility that these compounds alter not only epicardial cell function in the developing heart, but also block regeneration and transplantation of cells into a wounded adult heart.

描述（由申请人提供）：流行病学和实验室证据表明，TCDD等环境污染物会影响脊椎动物心脏的发育。 TCDD的受体是芳基烃受体（AHR），是配体激活的转录激活剂。我们使用斑马鱼，斑马鱼是研究体内心脏发育的理想系统，以记录对TCDD的心脏毒性反应。这些结果支持一个模型，其中TCDD破坏了发育中心的正常基因调节。我们的微阵列实验定义了TCDD在斑马鱼胚胎和成人心脏中产生的基因表达变化的模式。我们发现，AHR激活调节胚胎心脏细胞中与少年心脏细胞中不同的基因。成年心脏对TCDD心脏毒性具有抗性，除非在手术后再生失去的组织。胚胎和再生成人心脏中TCDD的敏感性与心外膜细胞迁移，分裂和分化对心脏至关重要的时期相关。这表明TCDD可能会影响心外膜细胞功能。我们开发了从胚胎斑马鱼心脏移植到受伤的成年肌肉中的方法。这些胚胎细胞被掺入受伤的心肌中。该项目的重点是以下问题：7心脏中AHR激活介导的TCDD的心毒性作用吗？这将通过使用转基因斑马鱼专门在心脏中操纵AHR活性来解决这一点。 7 TCDD是否会改变胚胎和再生成人心脏中的心外膜功能？这将通过确定TCDD是否阻断胚胎心脏中的心外膜细胞的形成和迁移来解决这。此外，我们将确定TCDD是否会改变成人心脏中胚胎样细胞的激活，迁移，生长和分化，以响应受伤。我们将比较成人受伤心脏中受TCDD影响的基因与胚胎心脏中受TCDD影响的基因。最后，我们将确定TCDD是否阻止将胚胎心脏细胞移植到受伤的斑马鱼心肌中。 7为什么TCDD的AHR激活会影响胚胎心脏中不同的基因，而不是在少年心脏或发育中的下巴中？我们将测试以下假设：不同组织中AHR复合物不同，在不同组织中产生不同的DNA结合位点特异性。作为替代方案，我们将检验以下假设：给定基因处的染色质结构可能允许胚胎心脏中的AHR结合，但是在其他组织中，该基因的染色质结构会有所不同，从而阻止AHR结合。这将使用与微阵列（芯片芯片分析）耦合的芯片测定法对此进行测试。公共卫生相关性：心脏缺陷是人类最常见的先天缺陷之一，最近的流行病学和实验室研究表明，心脏缺陷可能是由于暴露于2,3,7,8-四氯迪本佐 - 二苯甲酸 - 二恶英和多氯苯基的化学物质所引起的，分别为多苯基，通常是众所周知的二氧蛋白和PCB和PCB。该项目将基于斑马鱼Danio Rerio，这是一个重要的非人类模型系统，用于研究心脏发育。这项工作将确定有毒化合物破坏心脏发育的机制，并探讨这些化合物不仅会改变发育中心的心外膜细胞功能，而且会阻止细胞的再生和移植到受伤的成年心脏中。