Electron microscopy was used to quantify the subcellular distribution of the GLUT-1 isoform of the glucose transporter in developing microvessels of the brain of embryonic rats from E (embryonic stage) 13 to E19 and in adult rats. Gold-conjugated secondary antibodies were used to localize, on ultrathin sections of brain, a rabbit polyclonal antiserum (anti-GLUT-1) raised against a synthetic peptide encoding 13 amino acids of the C-terminus of the human glucose transporter. Staining was weak at E13 but increased in density during development into adulthood. The increase represented an increase in the absolute amount of transporter per vessel profile, with a concomitant decrease in vessel size with the narrowing of the wall. At early stages, the percentages of total particles per profile of lumenal membrane, ablumenal membrane, and cytoplasm were approximately equivalent. The ratio of lumenal to ablumenal particle density then shifted from below 1 at E13 to above 2 at E19 and to 4 in the adult. In contrast, vessels of the choroid plexus were devoid of labeling, but the choroid plexus epithelium stained as early as E15. In the brain, no astrocytes, neurons, or pericytes were stained at any stage examined. Developmental upregulation of the GLUT-1 glucose transporter therefore seems to occur at the blood-brain barrier, and the modulation of the subcellular distribution of the transporter can be correlated with other observed changes in the microvessels as they develop the blood-brain barrier phenotype.