title: Quantification of in vivo oxidative damage in Caenorhabditis elegans during aging by endogenous F3-isoprostane measurement.
abstract: 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.
authors.affiliation: University Medical Center Utrecht, Department of Metabolic Diseases and Netherlands Metabolomics Center, Utrecht, 3508 AB, The Netherlands.
authors.forename: Christiaan F
authors.initials: CF
authors.lastname: Labuschagne
authors.forename: Edwin C A
authors.initials: EC
authors.lastname: Stigter
authors.forename: Margriet M W B
authors.initials: MM
authors.lastname: Hendriks
authors.forename: Ruud
authors.initials: R
authors.lastname: Berger
authors.forename: Joshua
authors.initials: J
authors.lastname: Rokach
authors.forename: Hendrik C
authors.initials: HC
authors.lastname: Korswagen
authors.forename: Arjan B
authors.initials: AB
authors.lastname: Brenkman
mesh_terms: D000375:Aging / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D045303:Cytochromes b; D051858:Forkhead Transcription Factors; D015870:Gene Expression; D007328:Insulin / Q000235:genetics / Q000378:metabolism; D028421:Isoprostanes / Q000032:analysis / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D009154:Mutation; D010084:Oxidation-Reduction; D020675:Peroxisomes / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D017526:Receptor, IGF Type 1 / Q000235:genetics / Q000378:metabolism; D011972:Receptor, Insulin / Q000235:genetics / Q000378:metabolism; D011984:Sensory Receptor Cells; D013385:Succinate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: C104339:CLK-1 protein, C elegans; D029742:Caenorhabditis elegans Proteins; D051858:Forkhead Transcription Factors; D007328:Insulin; D028421:Isoprostanes; C472666:MEV-1 protein, C elegans; D017382:Reactive Oxygen Species; D014157:Transcription Factors; C108831:daf-16 protein, C elegans; D045303:Cytochromes b; D002374:Catalase; D013482:Superoxide Dismutase; D013385:Succinate Dehydrogenase; C107483:DAF-2 protein, C elegans; D017526:Receptor, IGF Type 1; D011972:Receptor, Insulin