Frum Tristan, Ralston Amy
Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, USA.
Methods Mol Biol. 2019;1893:335-352. doi: 10.1007/978-1-4939-8910-2_25.
The HIPPO signaling pathway plays an early and essential role in mammalian embryogenesis. The earliest known roles for HIPPO signaling during mouse development include segregating fetal and extraembryonic lineages and establishing the pluripotent progenitors of embryonic stem (ES) cells. In the mouse early embryo, HIPPO signaling responds to multiple cell biological inputs, including cell polarization, cytoskeleton, and cell environment, to influence gene expression and the first cell fate decisions in development. Methods to monitor and manipulate HIPPO signaling in the mouse early embryo are fundamental to discovering mechanisms regulating pluripotency in vivo, but properties of the early embryo, such as small cell number and spherical architecture, pose unique challenges for signaling pathway analysis. Here, we share approaches for visualizing HIPPO signaling in mouse early embryos. In addition, these methods can be applied to visualize HIPPO signaling in other spherical or cystic structures comprised of relatively few cells, such as organoids, or for the examination of other signaling pathways in these contexts.
HIPPO信号通路在哺乳动物胚胎发育过程中发挥着早期且至关重要的作用。在小鼠发育过程中，已知HIPPO信号通路最早的作用包括分离胎儿和胚外谱系以及建立胚胎干细胞（ES细胞）的多能祖细胞。在小鼠早期胚胎中，HIPPO信号通路对多种细胞生物学输入做出反应，包括细胞极化、细胞骨架和细胞环境，以影响基因表达和发育过程中的首个细胞命运决定。监测和操纵小鼠早期胚胎中HIPPO信号通路的方法对于发现体内调节多能性的机制至关重要，但早期胚胎的特性，如细胞数量少和球形结构，给信号通路分析带来了独特的挑战。在这里，我们分享在小鼠早期胚胎中可视化HIPPO信号通路的方法。此外，这些方法可应用于在其他由相对较少细胞组成的球形或囊性结构（如类器官）中可视化HIPPO信号通路，或用于在这些情况下检查其他信号通路。
