Techniques to Distinguish Apoptosis from Necroptosis.
Techniques to Distinguish Apoptosis from Necroptosis.
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DOI：
10.1101/pdb.top070375
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发表时间：
2016-04
影响因子：
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通讯作者：
M. Feoktistova;Fredrik Wållberg;T. Tenev;P. Geserick;M. Leverkus;P. Meier
中科院分区：
文献类型：
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作者：
M. Feoktistova;Fredrik Wållberg;T. Tenev;P. Geserick;M. Leverkus;P. Meier
细胞死亡的过程和控制细胞生长和增殖的过程一样受到严格的调控。最近对调节和执行细胞死亡的分子途径的研究发现，过多的信号级联导致不同的细胞死亡模式，包括“凋亡”、“坏死”、“自噬细胞死亡”和“有丝分裂灾难”。细胞可以很容易地从一种死亡形式切换到另一种形式；因此，拥有监测细胞正在经历的死亡形式的能力是至关重要的。有许多技术可以用来检测细胞死亡，当它们与敲除方法或特定信号通路的抑制剂(如caspase或RIP激酶途径)相结合时，它们可以快速剖析不同的细胞死亡途径。然而，揭示细胞死亡终点的技术不能重建导致死亡的事件序列；因此，它们需要补充能够区分所有形式的细胞死亡的方法。在体外培养条件下，凋亡细胞经常发生继发性坏死；因此，依赖高通量时间推移荧光视频显微镜的新方法是必要的，以提供对细胞死亡事件的时间分辨率。此外，可视化可以执行细胞凋亡或坏死性下垂的多蛋白信号中枢的组装有助于探索潜在的过程。在这里，我们介绍了一套可靠地区分坏死与凋亡和继发性坏死的技术，并使能够指导细胞凋亡或坏死性下垂的信号平台的研究成为可能。
The processes by which cells die are as tightly regulated as those that govern cell growth and proliferation. Recent studies of the molecular pathways that regulate and execute cell death have uncovered a plethora of signaling cascades that lead to distinct modes of cell death, including "apoptosis," "necrosis," "autophagic cell death," and "mitotic catastrophe." Cells can readily switch from one form of death to another; therefore, it is vital to have the ability to monitor the form of death that cells are undergoing. A number of techniques are available that allow the detection of cell death and when combined with either knockdown approaches or inhibitors of specific signaling pathways, such as caspase or RIP kinase pathways, they allow the rapid dissection of divergent cell death pathways. However, techniques that reveal the end point of cell death cannot reconstruct the sequence of events that have led to death; therefore, they need to be complemented with methods that can distinguish all forms of cell death. Apoptotic cells frequently undergo secondary necrosis under in vitro culture conditions; therefore, novel methods relying on high-throughput time-lapse fluorescence video microscopy are necessary to provide temporal resolution to cell death events. Further, visualizing the assembly of multiprotein signaling hubs that can execute apoptosis or necroptosis helps to explore the underlying processes. Here we introduce a suite of techniques that reliably distinguish necrosis from apoptosis and secondary necrosis, and that enable investigation of signaling platforms capable of instructing apoptosis or necroptosis.