Two vesicular glutamate transporters (VGluTs) have been identified at the molecular level very recently and revealed to possess similar pharmacological characteristics for glutamate uptake. Vesicular glutamate transporter 1 (VGluT1), which was originally named brain-specific Na+-dependent inorganic phosphate cotransporter (BNPI), is mainly expressed in telencephalic regions, whereas vesicular glutamate transporter 2 (VGluT2), formerly referred to as differentiation-associated Na+-dependent inorganic phosphate cotransporter (DNPI), is produced principally in diencephalic and lower brainstem regions. Since no other proteins show as high molecular similarity to VGluT1 or VGluT2 as the two transporters exhibit, it is likely that the mammalian central nervous system use only two gene products for vesicular glutamate uptake. Immunoelectron-microscopic analysis has revealed that the two VGluTs are located on synaptic vesicles in axon terminals making an asymmetric type of synapses, supporting that they serve as vesicular transporters in excitatory terminals. Furthermore, mRNA and immunoreactivity for VGluTs are distributed largely in a complementary fashion to distinct populations of excitatory neurons; for example, in the cerebral cortex, thalamocortical axon terminals use VGluT2, whereas excitatory axon terminals of corticocortical or intracortical fibers seem to apply VGluT1 for glutamate uptake. This complementary distribution might suggest that the two VGluTs have an as yet unknown difference in functions.