BACKGROUND The substantia nigra has been divided into three subdivisions. However, the cytoarchitecture of one of these subdivisions, the pars lateralis (SNl), has not been previously examined in detail at the light and electron microscopic levels in any species. In the adult opossum, the three nigral subdivisions can be easily distinguished as distinct, rostrocaudally oriented cell groups separated by neuron-free zones. Thus it was possible to determine the boundaries of the SNl unambiguously. This report covers the results of an examination of the morphology and organization of the SNl in the opossum. METHODS Material from 13 opossums was used for this study. Eight of the animals had been previously stained for Nissl substance (n = 4) or impregnated by the Golgi technique (n = 4). The remaining five animals were prepared for electron microscopic studies using standard procedures. RESULTS Two cell types were identified on the basis of morphological differences, small and medium-large neurons. Small neurons (10-18 microns long axis) have large nuclei with moderate amounts of heterochromatin and a thin rim of cytoplasm. They have long (up to 500 microns), spine-free dendrites. Medium-large neurons (18-54 microns long axis) have rounded nuclei with electron-lucent nucleoplasm. Few indentations of the nuclear envelope were observed. The surrounding cytoplasm has dense arrays of organelles. Nissl bodies are particularly prominent in the form of pyramids with their bases at juxtanuclear positions and their apices directed toward emerging dendrites. Dendrites of medium-large neurons are long (some > 1 mm in length), are primarily oriented in the frontal plane, and extend along the dorsal surface of or into the cerebral peduncle. Some cells have dendrites that are moderately spinous, whereas other neurons possess sparsely spinous dendrites. Relatively few synaptic profiles are observed to contact somata and proximal dendrites. CONCLUSIONS This report provides added morphological support for the idea that the SNl is a distinct subdivision of the substantia nigra, a distinction previously made on the basis of the physiologically characterized relationship between the lateral substantia nigra and orienting behaviors and seizure-related function.