The polysialylated neural cell adhesion molecule (PSA-NCAM) plays a role in axonal development and synaptic plasticity. Its pattern of expression is regulated temporally and topographically in the brain during development. However, it is unclear whether or not its subcellular location also changes. We have examined PSA-NCAM expression in relation to synapse formation in the developing rat striatum with immunohistochemistry and electron microscopy. Early in development, PSA-NCAM was present along the cytoplasmic membranes of neurons and in growth cones. PSA-NCAM expression became progressively confined to pre- and postsynaptic elements as neurons matured morphologically. Confirming previous results, a marked increase in the density of asymmetric synapses determined by using the physical dissector method was observed in the dorsolateral striatum between postnatal day 14 (P14) and P18. It was followed by a reduction between P18 and P25, when asymmetric synapse density reached adult levels. In contrast, the density of symmetric synapses had surpassed adult levels by P14. In the dorsomedial striatum, the density of asymmetric and symmetric synapses was similar at P18, at P25, and in adults. PSA-NCAM was associated with most asymmetric and symmetric synapses at P14 and P18 and was expressed in both pre- and postsynaptic elements of a majority (P14) or approximately half (P18) of the synapses. Most synapses lost PSA-NCAM expression between P18 and P25 in the dorsolateral striatum and between P25 and adult in the dorsomedial striatum. The data indicate that PSA-NCAM expression becomes restricted topographically during neuronal maturation but remains strategically associated with developing synapses during late postnatal development in the striatum.