保卫细胞中活性氧的实时活体监测

Real-Time In Vivo Monitoring of Reactive Oxygen Species in Guard Cells.

作者信息

Park Ky Young, Roubelakis-Angelakis Kalliopi A

机构信息

Department of Biology, Sunchon National University, 57922, Chonnam, South Korea.

Department of Biology, University of Crete, Voutes University Campus, 70013, Heraklion, Greece.

出版信息

Methods Mol Biol. 2018;1694:417-425. doi: 10.1007/978-1-4939-7398-9_34.

DOI:10.1007/978-1-4939-7398-9_34

PMID:29080184

Abstract

The intra-/intercellular homeostasis of reactive oxygen species (ROS), and especially of superoxides (O) and hydrogen peroxide (O) participate in signalling cascades which dictate developmental processes and reactions to biotic/abiotic stresses. Polyamine oxidases terminally oxidize/back convert polyamines generating HO. Recently, an NADPH-oxidase/Polyamine oxidase feedback loop was identified to control oxidative burst under salinity. Thus, the real-time localization/monitoring of ROS in specific cells, such as the guard cells, can be of great interest. Here we present a detailed description of the real-time in vivo monitoring of ROS in the guard cells using HO and O specific fluorescing probes, which can be used for studying ROS accumulation generated from any source, including the amine oxidases-dependent pathway, during development and stress.

摘要

活性氧（ROS），尤其是超氧化物（O）和过氧化氢（O）的细胞内/细胞间稳态参与信号级联反应，这些反应决定了发育过程以及对生物/非生物胁迫的反应。多胺氧化酶最终氧化/逆向转化多胺生成H₂O₂。最近，人们发现了一种NADPH氧化酶/多胺氧化酶反馈回路来控制盐胁迫下的氧化爆发。因此，在特定细胞（如保卫细胞）中对ROS进行实时定位/监测可能会非常有意义。在这里，我们详细描述了使用H₂O₂和O₂特异性荧光探针在保卫细胞中对ROS进行实时体内监测的方法，该方法可用于研究发育和胁迫期间从任何来源产生的ROS积累，包括胺氧化酶依赖性途径。