利用斑马鱼cloche模型研究BMP/GATA在心内膜发育中的作用

Heart valve is formed by the differentiation of the endocardial cells after atrioventricular cyclization metastasis. Deformed structural of valve will result in congenital heart disease, while the specific mechanism remains unknown..Zebrafish embryo is transparent and develops in vitro without relying on the circulation of the blood. Thus it serves as the ideal model of cardiovascular development. We have been screening a mutant of heart development defect, which mainly shows valve dysfunction. By hybrid screening we can build cloche model with GFP-labeled heart blood vessels and defected valve. In addition, we will clone this mutant gene from genome and explore the expression pattern of cloche. We will knock down cloche by morpholino and observe the structural of valve. Through gene silencing and overexpression technology, we can research the role of choche in the development of heart valve and related signal pathway..We can establish the animal model of cardiovascular developmental defects and study its mechanism, which may be benefit for the treatment and prevention of congenital heart disease.

心内膜是一层以内皮细胞为主，覆盖在心脏内表面连接心肌层的组织结构，对心脏瓣膜和房室间隔的形成至关重要，发育异常将导致房室间隔缺损和瓣膜功能异常。.斑马鱼胚胎透明体外发育，不依赖血液循环能够存活，是研究心血管发育的理想动物模型。前期工作利用ENU诱变技术筛选得到心脏发育缺陷突变体，主要表现为心脏瓣膜功能异常，通过与转基因斑马鱼杂交构建了心脏血管被特异标记的心内膜缺失的cloche模型，对该模型进行基因表达分析发现GATA和BMP表达下调。本项目将通过定位克隆cloche模型的突变基因并研究其在心内膜中的表达模式；通过morpholino技术敲减关键基因，观察瓣膜结构改变；采用基因沉默和过表达技术研究突变基因在心内膜发育中的作用，探索突变基因调控BMP和GATA的信号转导途径及其相互作用的分子机制。.建立心脏内膜发育缺陷的动物模型并研究其发生机制，能为先天性心脏病的预防和治疗提供理论依据。

心内膜是一层以内皮细胞为主，覆盖在心脏内表面连接心肌层的组织结构，对心脏瓣膜和房室间隔的形成至关重要，发育异常将导致房室间隔缺损和瓣膜功能异常。斑马鱼胚胎透明体外发育，不依赖血液循环能够存活，是研究心血管发育的理想动物模型。.利用ENU诱变技术成功构建出心脏被特异标记，心脏内膜发育异常表型稳定的斑马鱼cloche 模型；利用葡萄糖和乙醇协同诱导获得斑马鱼心包积液模型；建立了斑马鱼心血管发育干预小分子药物库，己经筛选到小分子化合物YSY2015能诱导转基因斑马鱼Tg(cmlc2: GFP)也成功获得cloche模型；检测了心脏发育调控信号新的基因cmlc2、has2、BMP-4、GATA-4和TBX5a在野生型和cloche 模型心脏发育过程的表达差异情况，进一步阐明新基因在心脏内膜发育中的分子机制。.建立心脏内膜发育缺陷的动物模型并研究其发生机制，能为先天性心脏病的预防和治疗提供理论依据。