Apoptosis and necrosis: Detection, discrimination and phagocytosis
Apoptosis and necrosis: Detection, discrimination and phagocytosis
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DOI：
10.1016/j.ymeth.2007.12.001
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发表时间：
2008-03-01
期刊：
影响因子：
4.8
通讯作者：
Vandenabeele, Peter
中科院分区：
文献类型：
--
作者：
Krysko, Dmitri V.;Vanden Berghe, Tom;Vandenabeele, Peter
细胞死亡的三种主要形态已被描述：细胞凋亡（I 型）、与自噬相关的细胞死亡（II 型）和坏死（III 型）。与自噬相关的细胞凋亡和细胞死亡可以通过某些生化事件来区分。然而，坏死的主要特征是缺乏 caspase 激活、细胞色素 c 释放和 DNA 寡核小体断裂。定义坏死的一个特别困难是，在没有吞噬作用的情况下，凋亡细胞变成具有许多原发性坏死形态特征的继发性坏死细胞。在这篇综述中，我们提出了一系列可用于识别坏死并将其与细胞凋亡区分开来的技术。这些技术依赖于以下细胞死亡参数：（1）形态学（延时和透射电子显微镜以及流式荧光细胞术）； (2) 细胞表面标志物（通过流式荧光细胞术测定磷脂酰丝氨酸暴露与膜通透性）； (3) 细胞内标记（流式荧光细胞术检测寡核小体 DNA 片段化、caspase 激活、Bid 裂解和蛋白质印迹检测细胞色素 c 释放）； (4)上清液中细胞外标志物的释放(caspases、HMGB-1和细胞角蛋白18)。最后，我们报告了可用于检查死亡细胞和吞噬细胞之间相互作用的方法。我们阐述了一种通过流式荧光细胞术检测死亡细胞吞噬作用的定量方法。我们还描述了一种最近开发的方法，该方法基于使用液相示踪剂和不同类型的显微镜、透射电子显微镜和荧光显微镜，来表征吞噬细胞内化垂死细胞的机制。 (C) 2008 Elsevier Inc. 保留所有权利。
Three major morphologies of cell death have been described: apoptosis (type I), cell death associated with autophagy (type II) and necrosis (type III). Apoptosis and cell death associated with autophagy can be distinguished by certain biochemical events. However, necrosis is characterized mostly in negative terms by the absence of caspase activation, cytochrome c release and DNA oligonucleosomal fragmentation. A particular difficulty in defining necrosis is that in the absence of phagocytosis apoptotic cells become secondary necrotic cells with many morphological features of primary necrosis. In this review, we present a selection of techniques that can be used to identify necrosis and to discriminate it from apoptosis. These techniques rely on the following cell death parameters: (1) morphology (time-lapse and transmission electron microscopy and flow fluorocytometry); (2) cell surface markers (phosphatidylserine exposure versus membrane permeability by flow fluorocytometry); (3) intracellular markers (oligonucleosomal DNA fragmentation by flow fluorocytometry, caspase activation, Bid cleavage and cytochrome c release by western blotting); (4) release of extracellular markers in the supernatant (caspases, HMGB-1 and cytokeratin 18). Finally, we report on methods that can be used to examine interactions between dying cells and phagocytes. We illustrate a quantitative method for detecting phagocytosis of dying cells by flow fluorocytometry. We also describe a recently developed approach based on the use of fluid phase tracers and different kind of microscopy, transmission electron and fluorescence microscopy, to characterize the mechanisms used by phagocytes to internalize dying cells. (C) 2008 Elsevier Inc. All rights reserved.