We have previously shown that the mature optic chiasm of a marsupial is divided morphologically into three regions, two lateral regions in which ipsilaterally projecting axons are confined and a central region containing only contralaterally projecting axons. By contrast, in the chiasms of eutherian (placental) mammals studied to date, there is no tripartite configuration. Ipsilaterally and contralaterally projecting axons from each eye are mixed in the caudal nerve and in each hemichiasm and encounter axons from the opposite eye near the midline of the chiasm. Here, we show that, unlike eutherians, marsupials have astrocytic processes in high concentrations in lateral regions of the nerve and rostral chiasm. Early in development, during the period when optic axons are growing through the chiasm, many intrachiasmatic cells are seen with densities five to eight times higher in lateral than in central chiasmatic regions. Such cells continue to be added to all chiasmatic regions; later in development, considerably more are added centrally, as the chiasm increases in volume. In the mature chiasm, cell densities are similar in all regions. By contrast to the marsupial, cell addition in the chiasm of a placental mammal, the ferret, is almost entirely restricted to later developmental stages, after axons have grown through the chiasm, and there are no obvious spatial variations in the distribution of cells during the period examined. During development, similar to the adult marsupial, ipsilaterally projecting axons do not approach the chiasmatic midline but remain confined laterally. We propose that the cells generated early and seen in high densities in the lateral chiasmatic regions of the marsupial may play a role in guiding retinal axons through this region of pathway selection. These data suggest that there is not a common pattern of developmental mechanisms that control the path of axons through the chiasm of different mammals.