Age-dependent survival, stress defense, and AMPK in Daphnia pulex after short-term exposure to a polystyrene nanoplastic

The widespread occurrence and accumulation of micro- and nanoplastics in aquatic environments has become a growing global concern. Generally, natural aquatic populations are characterized by a variety of multi-structured age groups, for which physiological and biochemical responses typically differ. The freshwater cladoceran, Daphnia pulex, is a model species used extensively in environmental monitoring studies and ecotoxicology testing. Here, the effects of a polystyrene nanoplastic on the physiological changes (i.e., survival) and expression levels of stress defense genes (i.e., those encoding antioxidant-mediated and heat shock proteins) in this freshwater flea were measured. Results from acute bioassays were used to determine the respective nanoplastic LC50 values for five age groups (1-, 4-, 7-, 14- and 21-day-old individuals): the obtained values for the 1- and 21-day-old D. pulex groups were similar (i.e., not significantly different). The expression levels of genes encoding key stress defense enzymes and proteins-SOD, CAT, GST, GPx, HSP70, and HSP90-were influenced by the nanoplastic in all the age groups, but not in the same way for each. Significant differences were observed among all age groups in their expression of the gene encoding the energy-sensing enzyme AMPK (adenosine monophosphate-activated protein kinase) α, β, and γ following exposure to the nanoplastic. Moreover, the expression of AMPK α was significantly increased in the 1-, 7-, and 21-day-old individuals exposed to nanoplastic relative to the control group. Together, these results indicate that age in D. pulex affects the sensitivity of its individuals to pollution from this nanoplastic, primarily via alterations to vital physiological and biochemical processes, such as cellular energy homeostasis and oxidation, which were demonstrated in vivo. We speculate that such age-related effects may extend to other nanoplastics and forms of pollution in D. pulex and perhaps similar marine organisms.