Betz cells are giant motoneurons located in layer Vb of the primate primary motor cortex. We conducted stereological analyses of Betz cells and neighboring pyramidal cells from the brains of six neurologically normal elderly humans to determine their volume, total number, and spatial distribution, and to relate these data to functional localization. The distribution of cellular volumes exhibits a bimodal pattern, delineating two different subpopulations. Betz cell volumes follow a mediolateral gradient, the largest Betz cells being located on the most medial part of the motor cortex. Additionally, the shape of Betz cells varies between the rostral and caudal parts of the primary motor cortex, supporting the notion that there are anatomically distinct zones in primary motor cortex. The total number of Betz cells per hemisphere accounts for about one-tenth of the total number of pyramidal cells in layer Vb. Analysis of spatial distribution using Voronoi tessellation revealed maximal clustering of Betz cells in a zone situated two-thirds from the midline along the mediolateral axis of the primary motor cortex. These data suggest that Betz cells have a discrete subregional distribution that may correspond to certain aspects of the functional parcellation of area 4. These results may offer a histological correlate of functional imaging studies and are relevant in the context of neurodegenerative diseases such as amyotrophic lateral sclerosis, progressive supranuclear palsy, and Guamanian amyotrophic lateral sclerosis/Parkinsonism-dementia, and in studies of normal brain aging. Anat Rec Part A 270A: 137-151, 2003. © 2003 Key words: amyotrophic lateral sclerosis; area 4; Betz cells; cytoarchitecture; human neocortex; motoneurons; stereologyThe human neocortex is characterized by regional and laminar specific distributions of a variety of neuronal subtypes and distinct afferent and efferent connections. The neocortex can be parcellated into a large number of more or less distinct fields according to microscopic architecture. Identification of the primary motor cortex and its boundaries, however, has been particularly contentious. Brodmann (1903Brodmann ( , 1909 originally described area 4 as an agranular zone, delineated by the presence of Betz cells, a subpopulation of giant infragranular pyramidal neurons in cortical layer V, located on its rostral border and buried in the depth of the anterior wall of the central sulcus. This definition has since been criticized (Zeki, 1979), and Brodmann's original delineation of the primary motor cortex has been revised by other investigators, who described an intermediate precentral area, an area precentralis A, area