Continuous tight junctions between vascular endothelial cells, the principal anatomical basis for the blood-brain barrier, have been investigated functionally and morphologically but their molecular components have not been defined. This communication reports that the protein ZO-1, a specific constituent of epithelial tight junctions, is found in human and rat brain vasculature. ZO-1-positive immunocytochemical staining forms a tightly banded pattern outlining individual endothelial cells in blood vessels of the human cerebral cortex. Rat brain exhibits a similar staining of blood vessels as well as ZO-1-positive staining around individual epithelial cells of the choroid plexus. The antiserum used for immunocytochemistry recognizes a protein of about 200 kDa in rat brain microvessels by Western blot. These findings indicate that ZO-1 is located at the interendothelial junctions of brain vasculature, implicating its importance as a component of the blood-brain barrier.