From Antioxidant Defenses to Transcriptomic Signatures: Concentration-Dependent Responses to Polystyrene Nanoplastics in Reef Fish

Nanoplastics (NPs) pose significant risks due to their small size and ability to penetrate biological tissues. However, the molecular pathways and cellular mechanisms affected by NP exposure in marine teleosts remain poorly understood, especially in tropical reef fishes. This study examined the impact of short-term (7 days) waterborne exposure of 100 nm-carboxyl-modified polystyrene NPs on the false clownfish (Amphiprion ocellaris) exposed at two daily concentrations: low (20 μg/L, environmentally relevant) and high (2000 μg/L). A multidisciplinary approach, including biochemical and transcriptomic analyses, was conducted to assess toxic effects. Biochemical assays revealed limited changes in antioxidant defenses (CAT, GR, GST, TOSC). However, the Integrated Biomarker Response index (IBRv2i) suggested a compromised physiological condition, supported by transcriptomic data. Transcriptomic profiling revealed 409 significantly differentially expressed genes (DEGs) in the high-concentration and 354 DEGs in the lowconcentration groups, with 120 shared DEGs mostly upregulated and indicative of a core molecular response. Collectively, the transcriptional profile of the low-concentration group resembled an early-warning, energy-reallocation strategy aimed at preserving essential sensory functions while minimizing expendable functions. The highconcentration group amplified the shared stress signature and recruited an additional 289 unique genes, resulting in pronounced enrichment of Gene Ontology terms related to “muscle contraction”, “oxygen transport”, “hydrogen-peroxide catabolism”, and “extracellular-matrix”. This study demonstrates that PS-NP exposure can alter gene expression and physiology in juvenile reef fish, even at environmentally relevant concentrations. Molecular responses varied with concentrations highlighting the role of exposure level in influencing biological systems and potential long-term impacts of NP pollution in marine environments.