Increased cell surface exposure of phosphatidylserine on propidium iodide negative thymocytes undergoing death by necrosis
Increased cell surface exposure of phosphatidylserine on propidium iodide negative thymocytes undergoing death by necrosis
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DOI：
10.1038/sj.cdd.4400540
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发表时间：
1999-07-01
影响因子：
12.4
通讯作者：
Beaver, J
中科院分区：
文献类型：
--
作者：
Waring, P;Lambert, D;Beaver, J
使用 FITC 标记的膜联蛋白-V 对碘化丙啶阴性细胞进行磷脂酰丝氨酸 (PS) 暴露，已用于定量体外和体内细胞凋亡。如果针对 PS 的标记膜联蛋白-V 在早期膜损伤后进入细胞，也可以检测正在坏死的细胞内的 PS。使用碘化丙啶可以将坏死或晚期凋亡细胞从流式细胞术分析中排除，碘化丙啶会进入膜完整性受损的细胞并对其进行染色。在这里，我们表明，仅因坏死而死亡的胸腺细胞在膜完整性丧失之前表现出早期的 PS 暴露。这种 PS 的早期暴露发生在用药物处理的细胞中，这些药物既能提高细胞内钙水平，又能与蛋白质硫醇基团相互作用。我们还证明，通过三种不同药物诱导发生凋亡的胸腺细胞中的 PS 暴露与钙升高无关，但与 DNA 断裂相关并先于 DNA 断裂。
Phosphatidylserine (PS) exposure on propidium iodide negative cells using FITC labelled annexin-V has been used to quantify apoptosis in vitro and in vivo. Detection of PS within cells undergoing necrosis is also possible if labelled annexin-V specific for PS enters the cell following early membrane damage. Necrotic or late apoptotic cells can be excluded from flow cytometric analysis using propidium iodide which enters and stains cells with compromised membrane integrity. Here we show that thymocytes undergoing death exclusively by necrosis show early exposure of PS prior to loss of membrane integrity. This early exposure of PS occurs in cells treated with agents which both raise intracellular calcium levels and are also capable of interacting with protein thiol groups. We also demonstrate that PS exposure in thymocytes induced to undergo apoptosis by three different agents does not correlate with calcium rises but correlates with and precedes DNA fragmentation.