Diverse subcellular locations of cryptogein-induced reactive oxygen species production in tobacco bright yellow-2 cells
Diverse subcellular locations of cryptogein-induced reactive oxygen species production in tobacco bright yellow-2 cells
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DOI：
10.1104/pp.106.090902
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发表时间：
2007-04-01
期刊：
影响因子：
7.4
通讯作者：
Fluhr, Robert
中科院分区：
文献类型：
--
作者：
Ashtamker, Cher;Kiss, Vladimir;Fluhr, Robert
活性氧（ROS）在许多细胞反应和信号通路中起着至关重要的作用，包括对病原体的氧化爆发防御反应。我们已经研究了在烟草（烟草）亮黄-2悬浮细胞中隐地蛋白诱导的ROS产生的非常早期的事件。使用Amplex Red和Amplex Ultra Red试剂，报告细胞群中实时H2 O2积累，我们表明H2 O2的内部信号比外部质外体信号发展得更快。还使用2 '，7'-二氯荧光素二乙酸酯荧光探针跟踪单个细胞中H2 O2的亚细胞积累。在内膜、细胞质和核隔室中检测到主要蓄积。当加入隐地蛋白时，信号首先在核区域产生，经过短暂的延迟后，在细胞周边产生。有趣的是，分离的细胞核能够以钙依赖的方式产生H2 O2，这意味着细胞核可以作为ROS产生的潜在活性源。这些结果表明，复杂的空间划分ROS积累和一个意想不到的时间序列的事件发生后，cryptogein应用，表明新的复杂性ROS信号级联。
Reactive oxygen species (ROS) play a crucial role in many cellular responses and signaling pathways, including the oxidative burst defense response to pathogens. We have examined very early events in cryptogein-induced ROS production in tobacco ( Nicotiana tabacum) Bright Yellow-2 suspension cells. Using Amplex Red and Amplex Ultra Red reagents, which report realtime H2O2 accumulation in cell populations, we show that the internal signal for H2O2 develops more rapidly than the external apoplastic signal. Subcellular accumulation of H2O2 was also followed in individual cells using the 2', 7'-dichlorofluorescein diacetate fluorescent probe. Major accumulation was detected in endomembrane, cytoplasmic, and nuclear compartments. When cryptogein was added, the signal developed first in the nuclear region and, after a short delay, in the cell periphery. Interestingly, isolated nuclei were capable of producing H2O2 in a calcium-dependent manner, implying that nuclei can serve as a potential active source of ROS production. These results show complex spatial compartmentalization for ROS accumulation and an unexpected temporal sequence of events that occurs after cryptogein application, suggesting novel intricacy in ROS-signaling cascades.