In vivo assessment of reactive oxygen species production and oxidative stress effects induced by chronic exposure to gamma radiation in Caenorhabditis elegans

Abstract

In the current study, effects of chronic exposure to ionizing gamma radiation were assessed in the radioresistant nematode Caenorhabditis elegans in order to understand whether antioxidant defences (AODs) could ameliorate radical formation, or if increased ROS levels would cause oxidative damage. This analysis was accompanied by phenotypical as well as molecular investigations, via assessment of reproductive capacity, somatic growth and RNA-seq analysis. The use of a fluorescent reporter strain (sod1::gfp) and two ratiometric biosensors (HyPer and Grx1-roGFP2) demonstrated increased ROS production (H₂O₂) and activation of AODs (SOD1 and Grx) in vivo. The data showed that at dose-rates ≤10 mGy h^-1 defence mechanisms were able to prevent the manifestation of oxidative stress. In contrast, at dose-rates ≥40 mGy h^-1 the continuous formation of radicals caused a redox shift, which lead to oxidative stress transcriptomic responses, including changes in mitochondrial functions, protein degradation, lipid metabolism and collagen synthesis. Moreover, genotoxic effects were among the most over-represented functions affected by chronic gamma irradiation, as indicated by differential regulation of genes involved in DNA damage, DNA repair, cell-cycle checkpoints, chromosome segregation and chromatin remodelling. Ultimately, the exposure to gamma radiation caused reprotoxic effects, with >20% reduction in the number of offspring per adult hermaphrodite at dose-rates ≥40 mGy h^-1, accompanied by the down-regulation of more than 300 genes related to reproductive system, apoptosis, meiotic functions and gamete development and fertilization.

Keywords: Caenorhabditis elegans; In vivo redox sensors; Ionizing gamma radiation; Mitochondrial dysfunction; Reactive oxygen species.