Zebrafish Cardiac Enhancer Trap Lines: New Tools for In Vivo Studies of Cardiovascular Development and Disease
Zebrafish Cardiac Enhancer Trap Lines: New Tools for In Vivo Studies of Cardiovascular Development and Disease
复制标题
DOI：
10.1002/dvdy.22203
复制
发表时间：
2010-03-01
影响因子：
2.5
通讯作者：
Korzh, Vladimir
中科院分区：
文献类型：
--
作者：
Poon, Kar-Lai;Liebling, Michael;Korzh, Vladimir
利用转座子介导的增强子陷阱（ET），我们获得了18个心脏增强子陷阱（CET）转基因斑马鱼品系。它们在确定的细胞类型（内膜、心肌和心外膜）或心脏的解剖区域（心房、心室、瓣膜或动脉球）中表现出EGFP表达。这些表达域中的大多数维持到成年。通过热不对称交错聚合酶链反应（TAIL-PCR）确定转座子插入的基因组位置。EGFP在一些CET中的表达模式是独特的，并且重现了转座子插入位点侧翼基因的表达。CET使我们能够使用快速扫描共聚焦显微镜结合图像重建来捕获体内胚胎心脏跳动的动态，及时（4D）制作三维电影，说明心脏收缩的区域特异性特征。这些CET细胞系代表了心脏发育、生理学和药物筛选体内研究的标记物工具箱。Developmental Dynamics 239：914-926，2010. (C)2010 Wiley-Liss，Inc.
Using the transposon-mediated enhancer trap (ET), we generated 18 cardiac enhancer trap (CET) transgenic zebrafish lines. They exhibit EGFP expression in defined cell types-the endocardium, myocardium, and epicardium-or in anatomical regions of the heart-the atrium, ventricle, valves, or bulbus arteriosus. Most of these expression domains are maintained into adulthood. The genomic locations of the transposon insertions were determined by thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR). The expression pattern of EGFP in some CETs is unique and recapitulates expression of genes flanking the transposon insertion site. The CETs enabled us to capture the dynamics of the embryonic heart beating in vivo using fast scanning confocal microscopy coupled with image reconstruction, producing three-dimensional movies in time (4D) illustrating region-specific features of heart contraction. This collection of CET lines represents a toolbox of markers for in vivo studies of heart development, physiology, and drug screening. Developmental Dynamics 239:914-926, 2010. (C) 2010 Wiley-Liss, Inc.