Abstract
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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 558-565 |
| Number of pages | 8 |
| Journal | Free Radical Biology and Medicine |
| Volume | 35 |
| Issue number | 6 |
| DOIs | |
| State | Published - Sep 15 2003 |
| Externally published | Yes |
Bibliographical note
Funding Information:This study was supported by the Brookdale National Fellowship (to M. S. G.), the Danish Research Agency (to A. O.), the Biotechnology and Biological Sciences Research Council (BBSRC) Special Studentship Program (to J. N. S.), NIH AG21069 and an Ellison Medical Foundation Senior Scholar Award (both to G. J. L.), and the Buck Institute (to A. O. and J. N. S.). The COPAS BIOSORT system was a generous gift from the Glenn Foundation for Medical Research and the Herbert Simon Foundation. All nematode strains were obtained from the Caenorhabditis Genetics Center, funded by the National Institutes of Health National Center for Research Resources. We would also like to thank Cliff Amend and the Benz Laboratory, Chris Lynch, and Andy Simpson. We thank the other members of the Lithgow Laboratory for helpful discussions.
Keywords
- Aging
- C. elegans
- Compound screen
- Free radicals
- Life span
- Superoxide dismutase/catalase mimetic
- Survival