In order to study the role of afferents in the maturation of cortical axons projecting from the ventrobasal thalamic complex (VB) to the barrel field (SI) cortex were labeled with the carbocyanine dye DiI, in aldehyde-fixed embryonic and newborn rat brains. Our results reveal that the first few thalamic axons are in the cortical plate by embryonic day (E) 19. Between E19 and the day of birth (E21 = PND 0), layers V and VI differentiate from the lower part of the cortical plate. On PND 0, a plexus of growth-cone tipped thalamic axons is present within the cortical plate and a few VB fibers have reached the marginal zone. Increasing numbers of thalamic afferents invade and ramify within the cortical plate on PND 1 and, over the course of the next 24 h, form a vibrissa-specific pattern in the lower part of this zone, prior to the differentiation of layer IV into a distinct lamina. This periphery-related organization is exhibited by VB afferents earlier than reported for other afferents to the cortex, by glia- or neuron-associated extracellular elements or by the cytoarchitectonic specializations (barrels) of stellate cells. Our observations, in conjunction with the previous studies, demonstrate that thalamic afferents may have a pivotal role in determining the morphological specification of the primary somatosensory cortex.