In recent years, plastic pollution has become increasingly serious, becoming global environmental concern. Plastics are widely used in daily life, and their addictive chemicals bisphenol A and phthalates continue to be produced and released into the environment in large quantities, due to improper disposal or degradation of plastics. Alarmingly high levels of bisphenol A and phthalates have been frequently detected in water and sediments and pose serious threats to the health of both animals and humans. Most current studies focus on their single effects on organisms, yet their combined effects on aquatic organisms are not clear. Given that these compounds often coexist in the natural environment, their combined effects deserve further investigation. Hence, the aim of this study was to evaluate the combined effects of plastic additives dibutyl phthalate (DBP) and bisphenol A (BPA) in aquatic organisms. To determine the effects of DBP and BPA on the early development and behavior in fish, in this study, using zebrafish (Danio rerio) as animal model, we investigated the single and combined effects of DBP and BPA on embryonic development, motor behavior and nervous system in zebrafish. Our results revealed that both individual and combined exposure to DBP and BPA resulted in decreased heart rate, shortened body length and accelerated hatching process in zebrafish embryos and larvae, but has no significant effect on total hatching rate and malformation rate. This suggested that DBP and BPA could interfere with normal physiological development. We observed that both individual and combined exposure to DBP and BPA enhanced the locomotor ability and activity level in zebrafish larvae. Furthermore, combined exposure to DBP and BPA increased the frequency of spontaneous tail curling in zebrafish embryos.We also found that DBP alone exposure resulted in increased malondialdehyde (MDA) levels and induced increased levels of apoptosis in brain cells. Combined exposure of DBP and BPA elevated the concentration of the oxidative stress product malondialdehyde (MDA), upregulated the transcriptional levels of genes related to nervous system development, reduced the neurotransmitter serotonin levels (5-HT), increased the brain cells apoptosis rates and the relative mRNA expression levels of the apoptosis-related gene bax. These findings provided strong evidence of the potential neurotoxic effects of DBP and BPA, suggesting that they may disrupt normal brain development and function in fish. In addition, our results indicated that DBP and BPA had antagonistic effects on the combined interference of early embryonic development (heart rate, hatching rate) and larval motor capacity (motor speed, distance, frequency of manic state, duration of manic state, frequency of active state, and duration of active state). The transcriptional levels of genes related to nervous system development (gfap, tph1b, th, syn2a and nestin) showed a certain synergistic effect. Our findings suggested that the abnormal locomotor behavior in zebrafish induced by combined exposure to DBP and BPA may be linked to changes in the mRNA expression of genes related to nervous system development and neurotransmitter levels. Our research provided new insights into the toxic effects of single and combined exposure to DBP and BPA at environmentally relevant concentrations on fish embryonic development, behavior, and the nervous system. These findings will help evaluate the ecological risk of DBP and BPA more objectively and precisely, and also can provide basic data for further study on combined disruptive mechanism of DBP and BPA on nervous system development in aquatic animals.