Laboratory for Physical Biology, RIKEN Center for Biosystems Dynamics Research, Kobe, Japan.
Faculty of Life and Environmental Sciences, University of Tsukuba, Tennodai, Ibaraki, Japan.
Methods Mol Biol. 2024;2828:37-43. doi: 10.1007/978-1-0716-4023-4_4.
Collective cell migration occurs when the orientation of cell polarity is aligned with each other in a group of cells. Such collective polarization depends on a reciprocal process between cell intrinsic mechanisms such as cell-cell adhesion and extracellular guidance mechanism such as wound healing and chemotaxis. As part of its development life cycle, individual single cells of Dictyostelium discoideum exhibit chemotaxis toward cAMP, which is secreted from a certain population of cells. During the formation of multicellular body by chemotaxis-dependent cell aggregation, D. discoideum is also known to relay on multiple cell-cell adhesion mechanisms. In particular, tail-following behavior at the contact site, called contact following of locomotion (CFL), plays a pivotal role on the formation of the multicellular body. However, whether and how CFL alone can lead to a formation of collective behavior was not well understood. KI cell is a mutant of D. discoideum that lacks all chemotactic activity. Yet, it can exhibit the CFL activity and show nontrivial collective cell migration. This mutant provides an excellent model system to analyze the mechanism of the CFL and the macroscopic phenomena brought by the CFL. This chapter describes protocols for using KI cell to understand the biophysics and cell biology behind the collective cell migration induced by CFL.
当一组细胞中的细胞极性方向彼此对齐时，就会发生集体细胞迁移。这种集体极化依赖于细胞内在机制（如细胞间黏附）和细胞外导向机制（如伤口愈合和趋化性）之间的相互作用过程。在其发育生命周期的一部分中，Dictyostelium discoideum 的单个单细胞表现出对 cAMP 的趋化性，cAMP 是由特定细胞群分泌的。在依赖趋化性的细胞聚集形成多细胞体的过程中，D. discoideum 还依赖于多种细胞间黏附机制。特别是在称为运动时的接触跟踪（CFL）的接触部位的尾随行为，在多细胞体的形成中起着关键作用。然而，CFL 本身是否以及如何导致集体行为的形成尚不清楚。KI 细胞是 D. discoideum 的突变体，缺乏所有趋化活性。然而，它可以表现出 CFL 活性并显示出重要的集体细胞迁移。该突变体提供了一个极好的模型系统，可用于分析 CFL 的机制以及由 CFL 引起的宏观现象。本章描述了使用 KI 细胞来理解 CFL 诱导的集体细胞迁移背后的生物物理学和细胞生物学的方案。
