Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA.
Department of Molecular Biology, Princeton University, Princeton, NJ, USA.
Methods Mol Biol. 2024;2805:187-201. doi: 10.1007/978-1-0716-3854-5_13.
Epidermal tissues are among the most striking examples of planar polarity. Insect bristles, fish scales, and mammalian fur are all uniformly oriented along an animal's body axis. The collective alignment of epidermal structures provides a valuable system to interrogate the signaling mechanisms that coordinate cellular behaviors at both local and tissue-levels. Here, we provide methods to analyze the planar organization of hair follicles within the mouse epidermis. Hair follicles are specified and bud into the underlying dermis during embryonic development. Shortly after, follicle cells dynamically rearrange to orient each follicle toward the anterior of the animal. When directional signaling is disrupted, hair follicles become misoriented. In this chapter, we describe how to create a spatial map of hair follicle orientations to reveal tissue-scale patterns in both embryonic and postnatal skin. Additionally, we provide a live imaging protocol that can be used to monitor cell movements in embryonic skin explants to reveal the cellular behaviors that polarize the hair follicle itself.
表皮组织是平面极性的最显著例子之一。昆虫的刚毛、鱼类的鳞片和哺乳动物的皮毛都是沿着动物的身体轴均匀排列的。表皮结构的集体排列为研究协调细胞行为的信号机制提供了一个有价值的系统,这些信号机制既可以在局部水平,也可以在组织水平上发挥作用。在这里,我们提供了分析小鼠表皮内毛囊平面组织的方法。毛囊在胚胎发育过程中被指定并发育成真皮下的芽。不久之后,毛囊细胞会动态地重新排列,使每个毛囊都朝向动物的前部。当定向信号被破坏时,毛囊就会发生错位。在这一章中,我们描述了如何创建毛囊取向的空间图谱,以揭示胚胎和出生后皮肤的组织尺度模式。此外,我们还提供了一个活体成像方案,可用于监测胚胎皮肤外植体中的细胞运动,以揭示使毛囊本身极化的细胞行为。