title: Using the MitoB method to assess levels of reactive oxygen species in ecological studies of oxidative stress.
abstract: In recent years evolutionary ecologists have become increasingly interested in the effects of reactive oxygen species (ROS) on the life-histories of animals. ROS levels have mostly been inferred indirectly due to the limitations of estimating ROS from in vitro methods. However, measuring ROS (hydrogen peroxide, H2O2) content in vivo is now possible using the MitoB probe. Here, we extend and refine the MitoB method to make it suitable for ecological studies of oxidative stress using the brown trout Salmo trutta as model. The MitoB method allows an evaluation of H2O2levels in living organisms over a timescale from hours to days. The method is flexible with regard to the duration of exposure and initial concentration of the MitoB probe, and there is no transfer of the MitoB probe between fish. H2O2levels were consistent across subsamples of the same liver but differed between muscle subsamples and between tissues of the same animal. The MitoB method provides a convenient method for measuring ROS levels in living animals over a significant period of time. Given its wide range of possible applications, it opens the opportunity to study the role of ROS in mediating life history trade-offs in ecological settings.
authors.affiliation: Institute of Biodiversity, Animal Health &Comparative Medicine, University of Glasgow, UK.
authors.forename: Karine
authors.initials: K
authors.lastname: Salin
authors.affiliation: Institute of Biodiversity, Animal Health &Comparative Medicine, University of Glasgow, UK.
authors.forename: Sonya K
authors.initials: SK
authors.lastname: Auer
authors.affiliation: Institute of Biodiversity, Animal Health &Comparative Medicine, University of Glasgow, UK.
authors.forename: Eugenia M
authors.initials: EM
authors.lastname: Villasevil
authors.affiliation: Institute of Biodiversity, Animal Health &Comparative Medicine, University of Glasgow, UK.
authors.forename: Graeme J
authors.initials: GJ
authors.lastname: Anderson
authors.affiliation: School of Chemistry, University of Glasgow, UK.
authors.forename: Andrew G
authors.initials: AG
authors.lastname: Cairns
authors.affiliation: Institute of Cardiovascular and Medical Sciences, University of Glasgow, UK.
authors.forename: William
authors.initials: W
authors.lastname: Mullen
authors.affiliation: School of Chemistry, University of Glasgow, UK.
authors.forename: Richard C
authors.initials: RC
authors.lastname: Hartley
authors.affiliation: Institute of Biodiversity, Animal Health &Comparative Medicine, University of Glasgow, UK.
authors.forename: Neil B
authors.initials: NB
authors.lastname: Metcalfe
mesh_terms: D000818:Animals; D004463:Ecology / Q000379:methods*; D006861:Hydrogen Peroxide / Q000032:analysis*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D009943:Organophosphorus Compounds / Q000008:administration & dosage*; D018384:Oxidative Stress; D010636:Phenols / Q000008:administration & dosage; D017382:Reactive Oxygen Species / Q000032:analysis*; D014337:Trout
chemical_list: C558697:MItoB compound; C558698:MItoP compound; D009943:Organophosphorus Compounds; D010636:Phenols; D017382:Reactive Oxygen Species; D006861:Hydrogen Peroxide