Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK.
Department of Haematology, Cambridge Institute for Medical Research, University of Cambridge, Cambridge, UK.
Nat Cell Biol. 2018 Feb;20(2):127-134. doi: 10.1038/s41556-017-0013-z. Epub 2018 Jan 8.
During gastrulation, cell types from all three germ layers are specified and the basic body plan is established . However, molecular analysis of this key developmental stage has been hampered by limited cell numbers and a paucity of markers. Single-cell RNA sequencing circumvents these problems, but has so far been limited to specific organ systems . Here, we report single-cell transcriptomic characterization of >20,000 cells immediately following gastrulation at E8.25 of mouse development. We identify 20 major cell types, which frequently contain substructure, including three distinct signatures in early foregut cells. Pseudo-space ordering of somitic progenitor cells identifies dynamic waves of transcription and candidate regulators, which are validated by molecular characterization of spatially resolved regions of the embryo. Within the endothelial population, cells that transition from haemogenic endothelial to erythro-myeloid progenitors specifically express Alox5 and its co-factor Alox5ap, which control leukotriene production. Functional assays using mouse embryonic stem cells demonstrate that leukotrienes promote haematopoietic progenitor cell generation. Thus, this comprehensive single-cell map can be exploited to reveal previously unrecognized pathways that contribute to tissue development.
在原肠胚形成过程中，所有三个胚层的细胞类型都被指定，并且建立了基本的身体计划。然而，这个关键发育阶段的分子分析受到细胞数量有限和标记物缺乏的阻碍。单细胞 RNA 测序解决了这些问题，但迄今为止仅限于特定的器官系统。在这里，我们报告了在 E8.25 时对发育中的小鼠进行的原肠胚形成后超过 20,000 个细胞的单细胞转录组特征。我们确定了 20 种主要的细胞类型，它们通常包含亚结构，包括早期前肠细胞中的三个不同的特征。体节祖细胞的伪空间排序确定了转录的动态波和候选调节剂，这些调节剂通过对胚胎空间分辨区域的分子特征验证。在内皮细胞群体中，从造血内皮细胞向红髓造血祖细胞过渡的细胞特异性表达 Alox5 和其共同因子 Alox5ap，它们控制白三烯的产生。使用小鼠胚胎干细胞的功能测定表明，白三烯促进造血祖细胞的生成。因此，这个全面的单细胞图谱可以被用来揭示以前未被识别的有助于组织发育的途径。
