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The properties of filopodia and spines and their association with the postsynaptic density (PSD) protein PSD-95 were studied during early development of cultured cortical neurons using time-lapse confocal microscopy. Neurons were transfected with recombinant PSD-95 constructs fused to green fluorescent protein (GFP) for, on average, either 8 d in vitro (DIV) or 14 DIV. We find that, during 1 hr of imaging, filopodia and spines bearing PSD-95/GFP clusters are significantly more stable (i.e., do not turnover) than those lacking clusters. When present within a spine precursor, a PSD-95/GFP cluster appeared to nucleate a relatively stable structure around which filopodium–spine membranes can move. Although processes bearing clusters were generally stable, in 8 DIV neurons, we observed that a subset (∼10%) of PSD-95/GFP clusters underwent rapid modular translocation between filopodia–spines and dendritic shafts. We conclude that, during early synaptic maturation, prefabricated PSD-95 clusters are trafficked in a developmentally regulated process that is associated with filopodial stabilization and synapse formation.

Modular Transport of Postsynaptic Density-95 Clusters and Association with Stable Spine Precursors during Early Development of Cortical Neurons