Background: Valproic acid (VPA), which is often used to treat epilepsy, causes a variety of neurobehavioral impairments that closely resemble the phenotype of autism spectrum disorder (ASD) in prenatally exposed individuals. Although the neurobehavioral eff, ects of extremely low concentrations of VPA have received limited research attention, several investigations have shown that the impact of VPA is connected with the concentration and exposure length. Objectives: In the current study, the aim was to fi, nd the lowest dose of VPA with the fewest side eff, ects to induce behavioral phenotypes related to ASD in zebrafi, sh. Methods: Zebrafi, sh embryos were fi, rst exposed to various concentrations of VPA (i. e., 1, 5, 15, 25, 48, and 75 , M) for 120 hours. Then, 42 days after conception, the survival rate, quality of hatching, and presence of deformity were assessed. Afterward, a 1 , M VPA was chosen based on observations, and behavioral experiments were carried out at 7, 21, and 42 days after fertilization (dpf). Additionally, 7dpf gene expression analysis was evaluated. Results: According to the obtained fi, ndings, behavioral abnormalities resembling ASD were induced in 7 and 21 dpf but not in 42 dpf after 120 hours of exposure to 1, M VPA. Real-time analysis in 7 dpf revealed signifi, cant changes in a number of genes linked to ASD, including lrp6, gsk3beta, chd8, and ctnnb. Conclusions: In conclusion, 120 hours of exposure of zebrafi, sh embryos to 1 , M of VPA might produce suitable VPA induces autism-like behavior models in zebrafi, sh larvae to research early and long-term neurobehavioral and gene expression alterations. Studies on drug development might adopt this approach.