The distribution of manganese-dependent superoxide dismutase, an enzyme that transforms superoxide radicals into hydrogen peroxide, was studied in the human brain post mortem using a sheep polyclonal antiserum raised against the enzyme from liver mitochondria. One band, corresponding to a protein of 22,000 mol. wt was detected in the human brain by western blot analysis. At the light-microscopy level, a punctate immunostaining was observed in the neuropil and in some but not all, glial and neuronal cell bodies. Electron-microscopy revealed that the staining was exclusively confined to the inner mitochondrial membrane. A heterogeneous distribution of manganese-dependent superoxide dismutase was observed in the human brain. In the forebrain, numerous immunostained neurons were detected in the striatum, thalamus, pallidal complex and the nucleus basalis of Meynert. In the cerebellum, only granular and Purkinje cells were immunostained. Various nuclei from the brainstem displayed superoxide dismutase immunoreactivity, including the cranial nerve nuclei, the nucleus supratrochlearis, the red nucleus, the substantia nigra, the nucleus cuneiformis and subcuneiformis, the nucleus parabigeminal, the nucleus centralis superior, the nucleus supraspinalis, the nucleus of the medullae oblongata and the gigantocellularis nucleus. Large pyramidal neurons containing superoxide dismutase were detected in the CA subsectors, the hilus of the hippocampus and the cerebral cortex. Smaller immunostained neurons were also observed in layers I, IV and VI of all cortical regions studied. The distribution of immunostained glial cells was more limited, and restricted to the internal and external capsules, the hypothalamus, the red nucleus, the pyramidal white matter and surrounding areas, the cerebral cortex and the sub-ependymal layer, the alveus and the stratum oriens of the hippocampus. This heterogeneous but not ubiquitous distribution of cells expressing manganese-dependent superoxide dismutase suggests that not all cells in the human brain are protected to the same extent against the deleterious effects of superoxide.