Quantification of in vivo oxidative damage in Caenorhabditis elegans during aging by endogenous F3‐isoprostane measurement

Aging Cell

Christiaan F. Labuschagne²  Edwin C. A. Stigter²  Margriet M. W. B. Hendriks²  Ruud Berger²  Joshua Rokach¹  Hendrik C. Korswagen³ 

[1] Claude Pepper Institute and Department of Chemistry, Florida Institute of Technology, Melbourne, FL, USA;University Medical Center Utrecht, Department of Metabolic Diseases and Netherlands Metabolomics Center, Utrecht, The Netherlands;Hubrecht Institute, Royal Academy of Arts and Sciences and University Medical Center Utrecht, Utrecht, The Netherlands

关键词: aging;    C. elegans;    F3‐isoprostanes;    Insulin/IGF‐1;    mitohormesis;    oxidative damage;    ROS;    SOD;   

DOI  :  10.1111/acel.12043

来源: Wiley

【 摘 要 】

Summary

Oxidative damage is thought to be a major cause in development of pathologies and aging. However, quantification of oxidative damage is methodologically difficult. Here, we present a robust liquid chromatography–tandem mass spectrometry (LC-MS/MS) approach for accurate, sensitive, and linear in vivo quantification of endogenous oxidative damage in the nematode Caenorhabditis elegans, based on F3-isoprostanes. F3-isoprostanes are prostaglandin-like markers of oxidative damage derived from lipid peroxidation by Reactive Oxygen Species (ROS). Oxidative damage was quantified in whole animals and in multiple cellular compartments, including mitochondria and peroxisomes. Mutants of the mitochondrial electron transport proteins mev-1 and clk-1 showed increased oxidative damage levels. Furthermore, analysis of Superoxide Dismutase (sod) and Catalase (ctl) mutants uncovered that oxidative damage levels cannot be inferred from the phenotype of resistance to pro-oxidants alone and revealed high oxidative damage in a small group of chemosensory neurons. Longitudinal analysis of aging nematodes revealed that oxidative damage increased specifically with postreproductive age. Remarkably, aging of the stress-resistant and long-lived daf-2 insulin/IGF-1 receptor mutant involved distinct daf-16-dependent phases of oxidative damage including a temporal increase at young adulthood. These observations are consistent with a hormetic response to ROS.

【 授权许可】

Unknown   
© 2012 The Authors Aging Cell © 2012 Blackwell Publishing Ltd/Anatomical Society of Great Britain and Ireland