Stowers Institute for Medical Research, 1000 East 50th Street, Kansas City, MO 64110, USA.
Development. 2014 Mar;141(5):1095-103. doi: 10.1242/dev.098855.
Embryonic cells that migrate long distances must critically balance cell division in order to maintain stream dynamics and population of peripheral targets. Yet details of individual cell division events and how cell cycle is related to phases of migration remain unclear. Here, we examined these questions using the chick cranial neural crest (NC). In vivo time-lapse imaging revealed that a typical migrating NC cell division event lasted ~1 hour and included four stereotypical steps. Cell tracking showed that dividing NC cells maintained position relative to non-dividing neighbors. NC cell division orientation and the time and distance to first division after neural tube exit were stochastic. To address how cell cycle is related to phases of migration, we used FACs analysis to identify significant spatiotemporal differences in NC cell cycle profiles. Two-photon photoconversion of single and small numbers of mKikGR-labeled NC cells confirmed that lead NC cells exhibited a nearly fourfold faster doubling time after populating the branchial arches. By contrast, Ki-67 staining showed that one out of every five later emerging NC cells exited the cell cycle after reaching proximal head targets. The relatively quiescent mitotic activity during NC cell migration to the branchial arches was altered when premigratory cells were reduced in number by tissue ablation. Together, our results provide the first comprehensive details of the pattern and dynamics of cell division events during cranial NC cell migration.
胚胎细胞在长距离迁移时必须严格平衡细胞分裂,以维持流态动力学和外周靶细胞的种群。然而,单个细胞分裂事件的细节以及细胞周期与迁移阶段的关系仍不清楚。在这里,我们使用鸡颅神经嵴(NC)研究了这些问题。体内延时成像显示,一个典型的迁移性 NC 细胞分裂事件持续约 1 小时,包括四个典型步骤。细胞跟踪显示,分裂的 NC 细胞相对于非分裂的相邻细胞保持位置不变。NC 细胞分裂方向以及神经管出口后第一次分裂的时间和距离是随机的。为了解细胞周期与迁移阶段的关系,我们使用 FACs 分析来鉴定 NC 细胞周期特征的显著时空差异。对单个和少量 mKikGR 标记的 NC 细胞进行双光子光转化证实,在分支弓中定居后,先导 NC 细胞的倍增时间几乎快了四倍。相比之下,Ki-67 染色显示,在到达近端头部靶标后,每五个后期出现的 NC 细胞中就有一个退出细胞周期。当通过组织消融减少前迁移细胞数量时,NC 细胞向鳃弓迁移过程中相对静止的有丝分裂活性发生改变。总的来说,我们的结果提供了颅神经嵴细胞迁移过程中细胞分裂事件的模式和动态的第一个全面细节。