Exploring real-time in vivo redox biology of developing and aging Caenorhabditis elegans
Exploring real-time in vivo redox biology of developing and aging Caenorhabditis elegans
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DOI：
10.1016/j.freeradbiomed.2011.11.037
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发表时间：
2012-03-01
影响因子：
7.4
通讯作者：
Braeckman, Bart P.
中科院分区：
文献类型：
--
作者：
Back, Patricia;De Vos, Winnok H.;Braeckman, Bart P.
活性氧（ROS）不再被认为仅仅是氧化代谢的有毒副产品。严格控制的 ROS 浓度和氧化还原电位的波动可能是信号传导过程的重要介质。要了解 ROS 和氧化还原状态在生理、应激反应、发育和衰老中的作用，需要对其在活体生物体中进行非破坏性、时空、实时量化。我们建立了带有基因编码荧光生物传感器 HyPer 和 Grx1-roGFP2 的秀丽隐杆线虫菌株，分别用于检测过氧化氢 (H2O2) 和谷胱甘肽氧化还原电位。尽管考虑到其透明度和遗传易处理性，秀丽隐杆线虫非常适合作为此类方法的模式生物，但它们以前从未在这种线虫中进行过尝试。我们发现 H2O2 处理明显诱导活体蠕虫中两种生物传感器的剂量依赖性、可逆反应。在胚胎后发育过程中，氧化型谷胱甘肽与还原型谷胱甘肽的比例下降。 H2O2 水平随着年龄的增长而增加，并且当通过饮食限制延长寿命时，这种效应会延迟。在年轻人中，我们检测到了几个具有独特氧化还原特性的区域，这些特性可能与其生物功能有关。我们的研究结果表明，基因编码的生物传感器可以揭示多细胞生物体内氧化还原生物学以前未知的细节。 (C) 2011 Elsevier Inc. 保留所有权利。
Reactive oxygen species (ROS) are no longer considered merely toxic by-products of the oxidative metabolism. Tightly controlled concentrations of ROS and fluctuations in redox potential may be important mediators of signaling processes. Understanding the role of ROS and redox status in physiology, stress response, development, and aging requires their nondisruptive, spatiotemporal, real-time quantification in a living organism. We established Caenorhabditis elegans strains bearing the genetically encoded fluorescent biosensors HyPer and Grx1-roGFP2 for the detection of hydrogen peroxide (H2O2) and the glutathione redox potential, respectively. Although, given its transparency and genetic tractability, C. elegans is perfectly suitable as a model organism for such approaches, they have never been tried before in this nematode. We found that H2O2 treatment clearly induces a dose-dependent, reversible response of both biosensors in the living worms. The ratio of oxidized to reduced glutathione decreases during postembryonic development. H2O2 levels increase with age and this effect is delayed when life span is extended by dietary restriction. In young adults, we detected several regions with distinct redox properties that may be linked to their biological function. Our findings demonstrate that genetically encoded biosensors can reveal previously unknown details of in vivo redox biology in multicellular organisms. (C) 2011 Elsevier Inc. All rights reserved.