Dendritic spines, small protrusions from dendritic shafts, receive most of the excitatory synapses in cortical regions. Spines are highly plastic structures that can be rapidly produced or lost in response to a wide array of internal and external stimuli, and they proliferate in acute slice preparations [J. Neurosci. 19 (1999) 2876]. The goal of the present study was to determine if protein synthesis is necessary for this spine proliferation. We found that the addition of protein synthesis inhibitors to acute slices (in which spines otherwise proliferate) blocked new spine growth. Furthermore, a population of longer spines was observed after 2 h but these did not develop during protein synthesis blockade. These data suggest that protein synthesis is necessary for new spine growth in acute brain slice preparations and support literature suggesting that newly produced spines develop from filopodia-like protrusions.