NLRP3 inflammasome activation is pivotal for cytokine secretion and pyroptosis in response to diverse stimuli, playing a crucial role in innate immunity. While extensively studied in mammals, the regulatory mechanisms governing NLRP3 activation in non-mammalian vertebrates remain largely unexplored. Teleosts, as basal vertebrates, represent an ideal model for exploring the evolutionary trajectory of inflammasome regulation. In this study, ABE assays, confocal microscopy, and biochemical analyses were applied to systematically characterize the mechanisms underlying NLRP3 inflammasome in teleosts, using large yellow croakers (