Kilon is a member of the IgLON family belonging to the immunoglobulin superfamily of cell adhesion molecules. In the present study, we investigated temporal and spatial changes of Kilon expression and its modulatory functions for synapse number using hippocampal cultured neurons. Kilon was observed to localize chiefly at axons and presynaptic terminals at early culture stage, however, it was seen mainly at dendritic postsynaptic spine of mature neurons at late culture stages. Kilon was solubilized with detergent treatment at early culture stages, while it resisted to extraction of the detergent in mature neurons. The overexpression of Kilon gene using a plasmid vector decreased the number of dendritic synapses at early culture stages, whereas the overexpression increased the number of dendritic synapses at late culture. These results demonstrate the alteration of modulatory function of Kilon for the number of dendritic synapses concomitant with changes in its localization and detergent solubility during neuronal culture development.