Genetic Analyst of Early Conduction System Development

早期传导系统开发的遗传分析

基本信息

批准号：
8316460
负责人：
Rima Arnaout
金额：
$ 5.77万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8316460
关键词：
Angiopoietins Animal Model Antibodies Arrhythmia Biological Assay Cardiac Cardiac conduction system Cardiomyopathies Cells Defect Development Dominant-Negative Mutation Electric Wiring Electrocardiogram Embryo Embryonic Development Embryonic Heart Endothelin Genetic Goals Heart Heart Diseases Human Imagery In Situ Hybridization Inherited Knowledge Link Location Maintenance Maps Microscopy Modeling Monitor Myocardial Myocardium Neuregulins Optics Organogenesis Pathway interactions Pattern Phenotype Play Process Research Research Training Role Signal Transduction Staging Staining method Stains Structure Sudden Death System Systems Development Testing Ventricular Ventricular Arrhythmia Ventricular Remodeling Work Zebrafish candidate marker cardiogenesis design and construction disease phenotype in vivo molecular marker mutant research study tissue regeneration tool

项目摘要

DESCRIPTION (provided by applicant): Understanding development of the specialized ventricular cardiac conduction system is critical to generating biologic treatments for arrhythmia and mastering cardiac tissue regeneration. Arrhythmia and sudden death characterize several cardiomyopathies with aberrant trabeculation or noncompaction. The development and maintenance of trabeculated ventricular myocardium is a key process in heart organogenesis and appears linked to the normal development of a mature ventricular cardiac conduction pattern by way of erbb2/neuregulin (nrg) signaling. The erbb2/nrg pathway is already known to play a part in conduction system development. However, neither the process of trabeculation nor its role in setting the stage for the developing conduction system is understood. The long-term goals of this research are to describe the early structure and function of the specialized conduction system, and to determine how trabeculation relates to conduction system development. The specific aims of this project are twofold. First, we will establish molecular markers of the specialized ventricular cardiac conduction system in zebrafish. We will test the hypotheses that the location of the first cells of the conduction system are located in the trabeculating myocardial layer, and that trabeculation mutants lack conduction system cells as assessed by analyzing the expression of these markers. Second, we will test the hypothesis that the process of trabeculation, and not just the nrg/erbb2 pathway, is required for conduction system development. The zebrafish is a useful model organism for these experiments due to its genetic tractability, translucent embryogenesis, and similarity to human electrophysiological phenotypes and disease. To identify conduction cells, we will use in situ hybridization and antibody staining of candidate markers on whole-mount and sectioned zebrafish hearts. To assess conduction system development in wild-type, mutant, and genetically altered hearts, we will use optical mapping and embryonic ECG monitoring. We will assess conduction phenotypes in erbb2 mutants, as well as in wild-type embryos injected with heart-specific dominant negative constructs designed to block other pathways involved in trabeculation. Studying the structure and function of the developing cardiac conduction system and its relationship to trabeculation will further our understanding of arrhythmia in both congenital and acquired cardiomyopathies and will provide tools and knowledge that can help treat ventricular arrhythmia and sudden death by manipulating conduction cells.

描述（由申请人提供）：了解专业心室心脏传导系统的发展对于产生心律不齐和掌握心脏组织再生的生物学治疗至关重要。心律不齐和突然死亡是多种心肌病的特征，这些心肌病具有异常的小偷或不合作。小径心肌的发育和维持是心脏器官发生的关键过程，似乎与ERBB2/Neuregulin（NRG）信号传导的成熟心室心脏传导模式的正常发展有关。众所周知，ERBB2/NRG途径在传导系统开发中起着作用。但是，小梁的过程或其在为发展传导系统设定阶段的作用均未理解。这项研究的长期目标是描述专门传导系统的早期结构和功能，并确定小梁的结构和功能与传导系统开发的关系。该项目的具体目标是双重的。首先，我们将在斑马鱼中建立专门的心室心脏传导系统的分子标记。我们将检验以下假设：传导系统的第一个细胞的位置位于小梁的心肌层中，而小梁沉积突变体缺乏通过分析这些标记物的表达来评估的传导系统细胞。其次，我们将检验以下假设：小径的过程，而不仅仅是NRG/ERBB2途径的传导系统开发所必需的。斑马鱼是这些实验的有用模型生物，由于其遗传性障碍性，半透明的胚胎发生以及与人类电生理学表型和疾病的相似性。为了鉴定传导细胞，我们将使用原位杂交和候选标记的原位杂交和斑马鱼心脏上的抗体染色。为了评估野生型，突变体和遗传改变的心脏中的传导系统开发，我们将使用光学映射和胚胎ECG监测。我们将评估ERBB2突变体中的传导表型，以及注入心脏特异性的主要负面构建体的野生型胚胎中，旨在阻止与小梁的其他途径。研究发育中的心脏传导系统的结构和功能及其与小梁的关系将进一步了解先天性和易术的心肌病中心律不齐的理解，并将提供工具和知识，以帮助治疗心室心律不齐和通过操纵暴导细胞的猝死。