By mediating cell-cell interactions, the neural cell adhesion molecule (N-CAM) has been implicated in various events such as axonal pathway formation, neurite outgrowth or synaptic remodelling. One mechanism by which N-CAM could contribute to these events has been proposed to involve modifications of the content of the molecule in polysialic acid. Here we have tested this possibility using an in vitro model of lesion-induced reactive synaptogenesis in hippocampal organotypic cultures. We present evidence that the sprouting reaction triggered by a section of CA3-CA1 connections in these cultures is associated with the expression of the highly sialylated form of N-CAM on regenerating neurites. In addition, we have examined the functional importance of this sialylation mechanism by analysing the effect of treating sectioned cultures with endo-neuraminidase-N which removes the polysialic acid portions of N-CAM. Measurements of the time course of recovery from the lesion, as assessed by the formation of new functional synaptic contacts across the section, showed that removal of the polysialic acid moieties of N-CAM significantly delays the sprouting reaction. The results support the idea that up regulations of highly sialylated forms of N-CAM are of functional importance in neurite sprouting and synapse regeneration in this in vitro model.