Individual cultured rat hippocampal neurons grown on glass coverslips were loaded with lucifer yellow (LY) and segments of their dendrites, containing dendritic spines, were exposed to the glutamate agonist N-methyl-D-aspartate (NMDA). Repeated pulse application of NMDA caused a reduction of spine length by 20%, within 1-2 h of exposure to the drug. No such changes were seen in spines exposed to NMDA in presence of the antagonist 2-aminophosphonovalerate or in spines on sister dendrites which were not exposed to NMDA. It is suggested that dendritic spines shrink following exposure to excessive synaptic stimulation.