Molecular characterization of thioredoxin reductase in waterflea Daphnia magna and its expression regulation by polystyrene microplastics

Highlights: • Daphnia magna thioredoxin reductase (TRxR) was revealed all the TRxR-features. • TRxR, arginine kinase and permease transcript were induced by microplastics (MPs). • Antioxidation, energy and material transportation played a positive role against MPs. -- Abstract: Global scale concerns regarding rise in microplastics pollution in the environment have recently aroused. Ingestion of microplastics by biota, including freshwater zooplankton has been well studied, however, despite keystone species in freshwater food webs, the molecular response (e.g. oxidative defense) of zooplankton in response to microplastics is still in its infancy. The thioredoxin (TRx) system has a vital function in cellular antioxidative defense via eliminating the excessive generation of reactive oxygen species (ROS). Therefore, it is necessary to investigate the effects of thioredoxin reductase (TRxR), due to its triggering the TRx catalysis cascade. The present study identified TRxR in Daphnia magna (Dm-TRxR) for the first time, and found that the full-length cDNA was 1862 bp long, containing an 1821-bp open reading frame. Homologous alignments showed the presence of conserved catalytic domain CVNVGC and the seleocysteine (SeCys) residue (U) located in the N- and C- terminal portions. Subsequently, the expression of Dm-TRxR, together with permease, arginine kinase (AK), was investigated by approach of quantitative real-time PCR after exposure to four (1.25-μm) polystyrene (PS) microbeads concentrations: 0 (control), 2, 4 and 8 mg L⁻¹ for 10 days. Dm-TRxR, permease and AK mRNA were significantly upregulated after exposure to 2, 4 mg L⁻¹ of PS, but then declined in the presence of 8 mg L⁻¹ PS. The gene expression results suggested that oxidative defense, energy production and substance extra cellular transportation were significantly regulated by microplastic exposure. Collectively, the present study will advance our knowledge regarding the biological effects of microplastic pollution on zooplankton, and builds a foundation for freshwater environmental studies on mechanistic and biochemical responses to microplastics.