Nanoparticles (NP) are extremely small particles, typically 1 to 100 nanometers in size. Due to their tiny scale, they exhibit unique properties, such as increased reactivity and size-dependent optical effects. Within the domain of nanoparticle research, Copper Oxide Nanoparticles (CuONPs) take center stage due to their special attributes. Several investigations have reported NP-induced oxidative stress responses, as well as additional pathophysiological consequences such as hepatotoxicity, genotoxicity, and nephrotoxicity. Probiotics are beneficial microorganisms that promote health, particularly in the gut. These live bacteria and yeasts offer a range of potential benefits, from aiding digestion to supporting the immune system. Probiotics can be found in certain foods, like yogurt, or taken as supplements. Therefore, in this study probiotics will be used for ameliorate the neurotoxicity if caused by CuONPs. In this study animals were categorized into 4 groups; Control group, CuONPs treated group, Probiotic and CuONPs + Probiotic. Different behavioral tests were performed to asses many neurobehavioral and muscular abnormality like pole test, open field test, learning ability, foot fault tests. A neurotransmitter such as acetylcholinesterase level was also evaluated. CuONPs treated groups showed alteration of behavioral parameters and significant reduction in neurotransmitters. However, when probiotic lactobasilus acidophilus were administered with CuONPs, behavioral parameters were return to normalization. There was a significant improvement in the neurotransmitters when probiotics were given along with CuONPs. Therefore, it can be concluded that CuONPs caused neurotoxicity at the tested dose and its amelioration can be done by consuming probiotic lactobasilus acidophilus.