The extracellular concentration of the excitatory neurotransmitter glutamate is kept low by the action of glutamate transporters in the plasma membranes of both neurons and glial cells. These transporters may play important roles, not only in the adult brain, but also in the developing brain, as glutamate is thought to modulate the formation and elimination of synapses as well as neuronal migration, proliferation and apoptosis. Here we demonstrate the developmental changes in the expression of two glutamate transporters, GLAST and GLT, by quantitative immunoblotting and by light and electron microscopic immunocytochemistry. At birth, GLT is not detectable, but GLAST is present at significant concentrations both in the forebrain and in the cerebellum. GLT is first detected in the forebrain and cerebellum in the second and third week, respectively. Both transporters reach adult levels by postnatal week 5. The development of the total glutamate uptake activity in the forebrain, as determined by solubilization and reconstitution of the transporters in liposomes, parallels that of GLT, in agreement with the observation that GLT is the predominant transporter in the adult brain. The regional distributions of both GLAST and GLT in the tissue are similar in young and adult rats. Only GLAST is detectable in the external germinal layer of the cerebellar cortex. Electron microscopical investigation demonstrated GLAST and GLT exclusively in glial cells in young as well as in adult animals.