We examined the subcellular distribution of tumor suppressor adenomatous polyposis coli (APC) in developing cultured rat hippocampal neurons using both immunofluorescent microscopy and immunoelectron microscopy. APC initially localized at the distal tips of all the cell processes. Between 12 and 24 h after plating, APC concentrated at the growth cone and in the distal portion of the longest process, which was growing very rapidly. The other processes, growing at a much slower rate, showed only weak immunoreactivity for APC. After maturation of the neurons, APC in the axons was restricted to the growth cones and distal portions. In contrast, APC formed a punctuated pattern along the dendritic processes. This subcellular distribution of APC was dependent on the organization of microtubules, but not actin filaments. Moreover, treatment of neurons with a PKCzeta specific inhibitor caused defects in the staining pattern of APC. These results suggest that APC may be involved in neuronal process extension, and that APC may exert different functions on axons and dendritic processes.