Lambert Lou, Danon Antoine
Institute de Biologie Paris Seine, UMR 7238, CNRS, Sorbonne Université, Paris, France.
Bio Protoc. 2024 Aug 5;14(15):e5043. doi: 10.21769/BioProtoc.5043.
Chlamydomonas () is a unicellular model alga that has been shown to undergo programmed cell death (PCD) that can be triggered in response to different stresses. We have recently shown that is particularly well suited to the study and quantification of PCD. We have shown for the first time that S-nitrosoglutathione (GSNO), a nitric oxide (NO) donor, is able to induce PCD and can be used as a study system in . In this article, we provide a simple and robust protocol for quantifying GSNO-induced PCD, which can be adapted to any other treatment. We explain how to detect NO production in the cell following GSNO treatment. We show how PCD can be identified simply by analyzing the degradation profile of genomic DNA. We also provide an easy and reproducible cell death quantification protocol, which makes it possible to follow the course of PCD over time and highlight very fine differences in the number of affected cells between different samples. Key features • Use of S-nitrosoglutathione (GSNO) as a means to study programmed cell death (PCD) in • Discrimination of PCD vs. necrosis. • In vivo determination of NO production in the cell. • A simple, robust protocol for PCD quantification.
衣藻()是一种单细胞模式藻类,已被证明会经历程序性细胞死亡(PCD),这种死亡可由不同应激触发。我们最近表明,衣藻特别适合用于PCD的研究和定量分析。我们首次表明,一氧化氮(NO)供体S - 亚硝基谷胱甘肽(GSNO)能够诱导PCD,并且可作为衣藻中的一种研究体系。在本文中,我们提供了一种简单且可靠的方案来定量GSNO诱导的PCD,该方案可适用于任何其他处理。我们解释了如何在GSNO处理后检测细胞中的NO生成。我们展示了如何通过分析基因组DNA的降解情况来简单地鉴定PCD。我们还提供了一种简便且可重复的细胞死亡定量方案,这使得能够随时间追踪PCD的进程,并突出不同样品之间受影响细胞数量的细微差异。关键特性 • 使用S - 亚硝基谷胱甘肽(GSNO)作为研究衣藻中程序性细胞死亡(PCD)的手段 • 区分PCD与坏死 • 细胞内NO生成的体内测定 • 一种简单、可靠的PCD定量方案