Cytotoxicity of acrylamide on neuronal cells cultured in a chemically defined medium was studied with morphological alteration, protein content, LDH activity and cumulative glucose consumption as toxicity indicators. The cumulative glucose consumption was significantly reduced by exposure to acrylamide before the other indicators were affected, suggesting that it would be the most sensitive indicator in the present study and that inhibition of glucose utilization might be one of the mechanisms of acrylamide neurotoxicity. The cumulative glucose consumption was applied for assessing cytotoxicity of acrylamide and its related compounds in neuronal cultures. The ED50 values were 0.8, 5.8, 15.0 mM for acrylamide, N-isopropylacrylamide, and methacrylamide, respectively, which are neurotoxic in in vivo studies. N,N'-methylene-bis-acrylamide, which is not reported to be neurotoxic, however, showed the lowest ED50 value, 0.2 mM, indicating that it was most potently toxic to neuronal cells. The results suggest the necessity of a cautious approach to neurotoxicity assessment from culture studies.