Using the MitoB method to assess levels of reactive oxygen species in ecological studies of oxidative stress

Salin, K., Auer, S. K., Villasevil, E. M., Anderson, G. J., Cairns, A. G., Mullen, W. , Hartley, R. C. and Metcalfe, N. B. (2017) Using the MitoB method to assess levels of reactive oxygen species in ecological studies of oxidative stress. Scientific Reports, 7, 41228. (doi: 10.1038/srep41228) (PMID:28117373) (PMCID:PMC5259740)

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 H2O2 levels 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. H2O2 levels 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.

• Item Type: Articles
• Additional Information: The authors acknowledge financial support from a European Research Council Advanced Grant (number 322784 to NBM).
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Cairns, Mr Andrew and Martin, Miss Eugenia and Hartley, Professor Richard and Anderson, Mr Graeme and Auer, Dr Sonya and Metcalfe, Professor Neil and Mullen, Dr Bill and Salin, Dr Karine
• Authors: Salin, K., Auer, S. K., Villasevil, E. M., Anderson, G. J., Cairns, A. G., Mullen, W., Hartley, R. C., and Metcalfe, N. B.
• College/School: College of Medical Veterinary and Life Sciences > School of Cardiovascular & Metabolic Health; College of Science and Engineering > School of Chemistry; College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine
• Journal Name: Scientific Reports
• Publisher: Nature Research
• ISSN: 2045-2322
• ISSN (Online): 2045-2322
• Published Online: 24 January 2017
• Copyright Holders: Copyright © 2017 The Authors
• First Published: First published in Scientific Reports 7: 41228
• Publisher Policy: Reproduced under a Creative Commons License