Ocean acidification induces neuronal hyperexcitation and anxiety-like behaviour in marine medaka via ASIC activation

Ocean acidification presents a significant threat to marine life, yet its neurobiological mechanisms remain poorly understood. This study examined how acid-sensing ion channels (ASICs) mediate neuronal excitability and anxiety-like behaviour in marine medaka (Oryzias melastigma) under elevated CO₂ concentrations (1000 and 1900 ppm). Transcriptomics revealed early upregulation of asic1a (4 days), while RT-qPCR demonstrated increased asic1a, asic1b, asic2 and asic4a (7 days), with only asic1a sustained at 30 days. Immunofluorescence confirmed heightened Asic2 in emotion-processing brain regions following acidification. Transmission electron microscopy unveiled distinct ultrastructural alterations: widened synaptic clefts, thinned postsynaptic densities, and decreased mitochondrial aspect ratios. Mitochondrial membrane potential assays revealed a reduction in membrane potential in response to acidification. Electrophysiological recordings showed increased neuronal firing count in the dorsolateral telencephalon under acidification, behavioural assessments revealed significant anxiety-like phenotypes, effects that were fully rescued by ASIC inhibition. These results indicated that temporal specificity in ASIC subtype expression in acidification response. The interplay of synaptic and mitochondrial dysfunction, neuronal hyperexcitability, and behavioural alterations suggested acidification impaired both synaptic transmission efficiency and mitochondrial function, destabilizing neural circuits. This study systematically elucidates the neurotoxic effects of ocean acidification on marine fish, providing critical scientific evidence for predicting the ecological impacts of climate change on marine organisms.