Gill Matthew S, Olsen Anders, Sampayo James N, Lithgow Gordon J
Buck Institute, Novato, CA 94945, USA.
Free Radic Biol Med. 2003 Sep 15;35(6):558-65. doi: 10.1016/s0891-5849(03)00328-9.
Many genetic or environmental manipulations that extend life span in the nematode Caenorhabditis elegans (C. elegans) also enhance survival following acute stresses such as oxidative damage and thermal stress. This coupling of stress response and aging mechanisms has proved a useful tool in identifying new genes that affect the aging process without the need for performing lengthy life span analyses. Therefore, it is likely that this approach may also be applied to the identification of pharmacological agents that extend life span through enhanced resistance to oxygen radicals or other stressors. To facilitate high-throughput drug screens in the nematode, we have developed a microtitre plate survival assay that uses uptake of the fluorescent dye SYTOX green as a marker of nematode death. An increase in throughput compared with the conventional survival assay was achieved by combining automated worm-handling technology with automated real-time fluorescence detection. We have validated this assay by examining survival during acute heat stress and protection against oxidative stress with the superoxide dismutase/catalase mimetic Euk-134. We propose that this novel method of survival analysis will accelerate the discovery of new pharmacological interventions in aging and oxidative stress.
许多延长线虫秀丽隐杆线虫（C. elegans）寿命的基因或环境操作，也能增强其在急性应激（如氧化损伤和热应激）后的存活率。应激反应与衰老机制之间的这种关联，已被证明是一种有用的工具，可用于识别影响衰老过程的新基因，而无需进行冗长的寿命分析。因此，这种方法很可能也适用于识别通过增强对氧自由基或其他应激源的抗性来延长寿命的药物制剂。为了便于在该线虫中进行高通量药物筛选，我们开发了一种微量滴定板存活测定法，该方法利用荧光染料SYTOX green的摄取作为线虫死亡的标志物。通过将自动化线虫处理技术与自动化实时荧光检测相结合，实现了与传统存活测定法相比通量的增加。我们通过检测急性热应激期间的存活率以及使用超氧化物歧化酶/过氧化氢酶模拟物Euk-134对氧化应激的保护作用，验证了该测定法。我们认为，这种新颖的存活分析方法将加速衰老和氧化应激方面新药理学干预措施的发现。
