Immunostaining for three neuronal proteins, nonphosphorylated neurofilament protein (with antibody SMI-32), calbindin, and parvalbumin, was used to examine the organization of layer IV in human primary visual cortex (area 17 or V1) specifically to determine whether, similar to the case in macaque V1, layer IVA is present and is divided into neurochemically distinct compartments. All three proteins are expressed by neurons that are unevenly distributed in layer IV of human V1; immunostaining for each protein includes a thin band corresponding to layer IVA of classic cytoarchitectonic studies. In this band, nonphosphorylated neurofilament protein immunoreactivity is present in relatively broad clusters of pyramidal cell somata and dendrites that appear as upwardly protruding parts of intense immunostaining in layer IVB, whereas immunoreactivity for calbindin and parvalbumin exists in somata of nonpyramidal neurons and in thin, dense clusters of punctate profiles. In tangential sections through layer IVA, the three proteins are seen in distinct compartments. Calbindin- and parvalbumin-immunostained neurons make up a thinly walled honeycomb or lattice, whereas neurons immunostained for nonphosphorylated neurofilament protein occupy the central lacunae. Direct comparison shows that neurons immunostained for calbindin occupy regions in layer IVA complementary to those immunostained for nonphosphorylated neurofilament protein. These data demonstrate a basic similarity in the organization of layer IV in macaques and humans. Layer IVA specifically is organized into complementary and neurochemically distinct compartments, including what appears to be a geniculocortically innervated and parvicellular-driven lattice and the interstitial lacunae formed by the periodic, upward protrusion of magnocellular-dominated layer IVB neurons.