Postsynaptic membrane trafficking plays an important role in synaptic plasticity, but the organization of trafficking machinery within dendritic spines is poorly understood. We use immunocytochemical analysis of rat hippocampal neurons to show that proteins mediating endocytosis are systematically arrayed within dendritic spines, tangential to the synapse. Thus, previously unrecognized lateral domains of the spine organize endocytic protein machinery at sites removed from the postsynaptic density.