Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, United States of America.
Department of Biology, Stanford University, Stanford, California, United States of America.
PLoS Biol. 2019 Jul 17;17(7):e3000381. doi: 10.1371/journal.pbio.3000381. eCollection 2019 Jul.
The primary cilium is a central signaling hub in cell proliferation and differentiation and is built and disassembled every cell cycle in many animal cells. Disassembly is critically important, as misregulation or delay of cilia loss leads to cell cycle defects. The physical means by which cilia are lost are poorly understood but are thought to involve resorption of ciliary components into the cell body. To investigate cilium loss in mammalian cells, we used live-cell imaging to comprehensively characterize individual events. The predominant mode of cilium loss was rapid deciliation, in which the membrane and axoneme of the cilium was shed from the cell. Gradual resorption was also observed, as well as events in which a period of gradual resorption was followed by rapid deciliation. Deciliation resulted in intact shed cilia that could be recovered from culture medium and contained both membrane and axoneme proteins. We modulated levels of katanin and intracellular calcium, two putative regulators of deciliation, and found that excess katanin promotes cilia loss by deciliation, independently of calcium. Together, these results suggest that mammalian ciliary loss involves a tunable decision between deciliation and resorption.
初级纤毛是细胞增殖和分化的核心信号枢纽,在许多动物细胞中,每个细胞周期都会构建和拆卸。拆卸至关重要,因为纤毛的调节或延迟丢失会导致细胞周期缺陷。纤毛丢失的物理机制尚不清楚,但据认为涉及纤毛成分被吸收到细胞体中。为了研究哺乳动物细胞中的纤毛丢失,我们使用活细胞成像技术全面描述了单个事件。纤毛丢失的主要模式是快速去纤毛,其中纤毛的膜和轴丝从细胞上脱落。也观察到逐渐吸收的情况,以及在逐渐吸收之后快速去纤毛的情况。去纤毛导致完整的脱落纤毛,可以从培养基中回收,并且包含膜和轴丝蛋白。我们调节了 katanin 和细胞内钙的水平,这两种纤毛去化的潜在调节剂,发现过量的 katain 可通过去纤毛促进纤毛丢失,而与钙无关。这些结果表明,哺乳动物纤毛丢失涉及到去纤毛和吸收之间的可调节决策。