Aix-Marseille Université, CNRS UMR 7288, IBDM, Marseille, France.
Department of Genetics, Albert Einstein College of Medicine, Bronx, NY, USA.
Methods Mol Biol. 2022;2438:231-250. doi: 10.1007/978-1-0716-2035-9_15.
Epithelial cardiac progenitor cells of the second heart field (SHF) contribute to growth of the vertebrate heart tube by progressive addition of cells from the dorsal pericardial wall to the cardiac poles. Perturbation of SHF development, including defects in apicobasal or planar polarity, results in shortening of the heart tube and a spectrum of congenital heart defects. Here, we provide detailed protocols for fixed section and wholemount immunofluorescence and live imaging approaches to studying the epithelial properties of cardiac progenitors in the dorsal pericardial wall during mouse heart development. Whole-embryo culture and electroporation methods are also presented, allowing for pharmacological and genetic perturbation of SHF development, as well as image analysis approaches to quantify cell features across the progenitor cell epithelium. These protocols are broadly applicable to the study of epithelia in the early embryo.
第二心脏场（SHF）的心脏上皮祖细胞通过从背侧心包壁向心脏极渐进式地添加细胞来促进脊椎动物心脏管的生长。SHF 发育的干扰，包括顶端-基底或平面极性的缺陷，导致心脏管缩短和一系列先天性心脏缺陷。在这里，我们提供了详细的固定切片和全胚胎免疫荧光和活体成像方案，用于研究心脏发育过程中背侧心包壁中心脏祖细胞的上皮特性。还介绍了整个胚胎培养和电穿孔方法，可用于 SHF 发育的药理学和遗传学干扰，以及图像分析方法，用于跨祖细胞上皮定量细胞特征。这些方案广泛适用于早期胚胎中上皮的研究。
