This paper examines both the anatomical changes in the distribution of vimentin intermediate filament protein and the biochemical changes in vimentin and its degradative enzyme during postnatal brain development in the tree shrew (Tupaia belangeri). A pattern of multiple immunoreactive bands at birth (postnatal day 0, or P0) was revealed in nitrocellulose blots of polyacrylamide gels ("Western blots"). These multiple bands gradually disappear during development, and in the adult a single band at the published molecular weight for vimentin (57 kD) is seen. This pattern of bands probably reflects shifts in the activity of a calcium-activated vimentin protease. The changes in the anatomical distribution of vimentin-immunoreactive (vimentin+) cells and their fine processes parallel the biochemical shifts seen in immunoblots. We have examined the neocortex, lateral geniculate nucleus (LGN), and hippocampus in detail. During the first postnatal week, vimentin+ glia, especially radial glia, are prominent in both neocortex and hippocampus. In contrast, only a few vimentin+ radial glia remain in the thalamus at this age. Vimentin+ glia appear to coincide with bundles of axons and often seem to outline subdivisions of thalamic nuclei. Additionally, cellular layers of the lateral geniculate nucleus (LGN) appear to stain with antibodies to vimentin several days before the characteristic neuronal cell layers appear in this area. During the second postnatal week, vimentin+ cells appear in "patches" throughout the cortex. Some subdivisions of the thalamus and hippocampus (as defined by cytoarchitectonic differences in the adult) are distinct when the tissue is stained with an antibody to vimentin, even though a conventional Nissl stain at this age shows no apparent delineation in these same regions. Finally, in the adult, only a few vimentin+ cells remain, primarily in the white matter. Taken together, these results suggest that the remodeling of vimentin+ intermediate filaments in immature glial cells (including radial glia) is paralleled by the action of the enzyme which breaks down these filaments. The apparent activity of this enzyme is high early in development as radial and other glia are rapidly dividing and undergoing morphological changes, with a decrease in activity in the juvenile and adult brain, as immature glial cells are supplanted by mature forms.