Eukaryotic high-affinity glutamate transporters (also called excitatory amino acid transporters, EAATs) contain five isoforms, GLAST (EAAT1), GLT-1 (EAAT2), EAAC1 (EAAT3), EAAT4 and EAAT5. Structural analysis reveal the membrane topology of glutamate transporters, the structural differences between the eukaryotic and prokaryotic glutamate transporters, and the binding sites for substrate and coupled ions during the transport cycle. Functional studies indicate that EAATs are involved in synaptic transmission, and EAATs play key roles in avoiding glutamate excitotoxicity. Besides, EAATs also participate in regulating learning, memory and motor behavior. In this review, combined with our previous work, we summarize the progress in the research of the structure and function of high-affinity glutamate transporters recently.