Adequate alternatives to conventional animal testing are needed to study developmental neurotoxicity (DNT). Here, we used kinematic analysis to assess DNT of known (toluene (TOL) and chlorpyrifos (CPS)) and putative (β-N-methylamino-L-alanine (BMAA)) neurotoxic compounds. Drosophila melanogaster was exposed to these compounds during development and evaluated for survival and adult kinematic parameters using the FlyWalker system, a kinematics evaluation method. At concentrations that do not induce general toxicity, the solvent DMSO had a significant effect on kinematic parameters. Moreover, while TOL did not significantly induce lethality or kinematic dysfunction, CPS not only induced developmental lethality but also significantly impaired coordination in comparison to DMSO. Interestingly, BMAA, which was not lethal during development, induced motor decay in young adult animals, phenotypically resembling aged flies, an effect later attenuated upon aging. Furthermore, BMAA induced abnormal development of leg motor neuron projections. Our results suggest that our kinematic approach can assess potential DNT of chemical compounds.