A light and electron microscopic study of the developing corpus callosum was carried out in foetal and neonatal rats in order to determine the mode of growth of the earliest callosal axons across the midline and to investigate the potential role played by non-neuronal cells during the formation of the tract. The axons of the corpus callosum first cross the midline between the 18th and 19th days of gestation by traversing the anterodorsal aspect of the pre-existing hippocampal commissure. Prior to the appearance of the callosal axons at the midline, there is an aggregation of astrocyte processes anterior and dorsal to the hippocampal commissure. Careful examination of these processes in different planes of section shows that they are not organized in any obvious way that would provide a clearly defined path for the growing axons; nor are there any preferentially oriented extracellular spaces at the midline. No specialized membrane contacts could be seen between non-neuronal cell processes and the early callosal axons. Thus, there is no overt morphological evidence for an active role of non-neuronal cells in axon guidance in the initial formation of the corpus callosum. The development of the corpus callosum is accompanied by the formation of a temporary cavum septi pellucidi, which is always closed to the subarachnoid space. The cavum persists during the first postnatal week, after which time it becomes populated by cells of the lateral septal nuclei. Macrophages are present within the cavum and may play a role in its formation. Macrophages are also found within parts of the corpus callosum. No obvious degeneration of axons is seen in the corpus callosum during its early development.