An automated high-throughput assay for survival of the nematode Caenorhabditis elegans
An automated high-throughput assay for survival of the nematode Caenorhabditis elegans
复制标题
DOI：
10.1016/s0891-5849(03)00328-9
复制
发表时间：
2003-09-15
影响因子：
7.4
通讯作者：
Lithgow, GJ
中科院分区：
文献类型：
--
作者：
Gill, MS;Olsen, A;Lithgow, GJ
许多遗传或环境操作，延长线虫秀丽隐杆线虫（C elegans）的寿命，也提高生存急性应激，如氧化损伤和热应激。这种应激反应和衰老机制的耦合已被证明是一种有用的工具，可以识别影响衰老过程的新基因，而无需进行漫长的寿命分析。因此，这种方法也可能适用于鉴定通过增强对氧自由基或其他应激源的抵抗力来延长寿命的药理学试剂。为了促进线虫中的高通量药物筛选，我们开发了微量滴定板存活测定，其使用荧光染料SYTOX绿色的摄取作为线虫死亡的标记。通过将自动化蠕虫处理技术与自动化实时荧光检测相结合，实现了与常规存活测定相比的吞吐量增加。我们已经验证了这种测定方法，通过检查急性热应激和保护对氧化应激与超氧化物歧化酶/过氧化氢酶模拟物Euk-134的生存。我们认为，这种新的生存分析方法将加速发现新的药物干预衰老和氧化应激。(C)2003年爱思唯尔。
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. (C) 2003 Elsevier.