It remains challenging to construct a complete cell lineage map of the origin of vascular endothelial cells in any vertebrate embryo. Here, we report the application of in toto light-sheet fluorescence imaging of embryos to trace the origin of vascular endothelial cells (ECs) at single-cell resolution in zebrafish. We first adapted a previously reported method to embryo mounting and light-sheet imaging, created an alignment, fusion, and extraction all-in-one software (AFEIO) for processing big data, and performed quantitative analysis of cell lineage relationships using commercially available Imaris software. Our data revealed that vascular ECs originated from broad regions of the gastrula along the dorsal–ventral and anterior–posterior axes, of which the dorsal–anterior cells contributed to cerebral ECs, the dorsal–lateral cells to anterior trunk ECs, and the ventral–lateral cells to posterior trunk and tail ECs. Therefore, this work, to our knowledge, charts the first comprehensive map of the gastrula origin of vascular ECs in zebrafish, and has potential applications for studying the origin of any embryonic organs in zebrafish and other model organisms.

构建任何脊椎动物胚胎中血管内皮细胞起源的完整细胞谱图仍然具有挑战性。在这里，我们报道了在to-to-to-sheet胚胎荧光成像中的应用，以追踪斑马鱼中单细胞分辨率的血管内皮细胞（EC）的起源。我们首先将先前报道的方法应用于胚胎安装和光片成像，创建了用于处理大数据的对齐，融合和提取多合一软件（AFEIO），并使用市售的Imaris进行了细胞谱系关系的定量分析软件。我们的数据显示，血管内皮细胞起源于沿腹-腹和前-后轴的胃大片区域，其中背-前细胞促成脑内皮细胞，背侧细胞至前躯干EC，腹侧细胞至后躯干和尾EC。因此，据我们所知，这项工作绘制了斑马鱼中血管EC胃下胃的第一个综合图，并具有潜在的用途，可用于研究斑马鱼和其他模型生物中任何胚胎器官的起源。