The somata of corticospinal neurons were labeled with horseradish peroxidase that had been applied to a hemisection of the spinal cord at the C1-C2 junction in 22 species of mammals. After tetramethylbenzidine processing, with and without counterstaining with cresyl violet or neutral red, the labeled cells in systematic sets of sections throughout the cerebral cortex were plotted and counted. Several morphological features of the corticospinal cells were examined including their cell type, number, density, concentration, laminar distribution, and their distribution across the cortical surface. The results show that the labeled corticospinal neurons were invariably layer V pyramidal cells. However, in many mammals they were found to be stacked one above the other within layer V, sometimes many neurons deep. Despite the concentration of corticospinal neurons within layer V, many unlabeled neurons were also present within the layer throughout the extent of the labeled region. The results also indicate that at least two spatially distinct regions of neocortex originate corticospinal fibers in each of the animals in the sample. In addition to these two regions, a third segregated region is present in the cortex of primates and an apparently different third region is present in the cortex of Glires (Rodentia and Lagomorpha). The third region of corticospinal cortex in primates is located on the lateral surface of the cortex in prosimians and New World monkeys and is buried in the caudal bank of the inferior arcuate sulcus in Old World monkeys. The results also show a predominantly contralateral corticospinal tract in all but 4 of the 22 mammals in the sample. Although these 4 mammals are each members of the order Insectivora, a less modified member of the same order possessed the predominantly contralateral projection of most mammals, hence denying the notion that a predominantly ipsilateral tract is a characteristic of Insectivora.