The cytochrome oxidase (CO) blobs and interblobs in layer 3B of primate visual cortex have different sets of corticocortical connections. Cortical layers below layer 3B also project corticocortically, but the relationship of efferent projections from the deeper layers to the overlying blob/interblob architecture is less clear. We studied the tangential organization of neurons projecting from primary visual cortex (V1) to the middle temporal visual area (MT) and their relationship to the CO blobs. MT-projecting neurons in two primate species, bush babies and owl monkeys, were retrogradely labeled, then charted in tangential sections, and compared to the positions of the overlying CO blobs. In both primate species, MT-projecting neurons in layer 3C were unevenly distributed in the tangential plane, with dense patches of labeled cells that were aligned with the CO blobs. A novel two-dimensional spatial correlation method was used to show the colocalization of MT-projecting cells with the overlying blobs. Chi-square analyses performed with the cortical surface equally divided into compartments of blob, interblob, and blob/interblob borders showed that blob columns tended to have about 1.5 times more MT-projecting cells (P < 0.0001) than interblob columns. Similar analyses were applied to published data on V1 cells projecting to area MT in macaque monkey (Shipp and Zeki [1989] Euro J Neurosci 1:310-332). Again, the results showed a significant correlation between the cell distribution and CO blobs. Taken together, these results suggest that layer 3C is not uniform but is made up of a mosaic of cells that project to area MT and cells that project to some other location. These findings also indicate that the mosaic organization of layer 3C is related in some unique way to the overlying CO architecture.