In this study, we used the adult zebrafish model of ACR acute neurotoxicity for determining the suitability of NAC to protect the brain against ACR toxicity. First, the potential protective effects of NAC on the ACR neurotoxicity were evaluated at the behavioral and molecular (transcriptome and proteome) levels. Then, in order to understand the mild to negligible protective effect of NAC, the ability of this drug to cross BBB by passive diffusion was evaluated by different approaches. Whereas the BBB parallel artificial membrane permeability assay (BBB-PAMPA) was used to determine the ability of NAC to cross BBB-like phospholipid membranes by passive diffusion, the determination of the NAC content in the brain provides direct evidences of the BBB permeability in vivo. Finally, we evaluated the ability of drug to recover the depleted GSx stores and to scavenge ACR in the brain of ACR-exposed fish.