The kinase mammalian target of rapamycin (mTOR) is stimulated by insulin, growth factors and nutrients and confers survival of several cell types. The kinase has previously been shown to stimulate amino acid uptake. In neurons the cellular uptake of glutamate by the excitatory amino acid transporters (EAATs) decreases excitation and thus confers protection against excitotoxicity. The present study explored, whether the wild-type mTOR regulates the EAAT3 (SLC1A1). To this end, cRNA encoding the EAAT3 was injected into Xenopusoocytes with or without cRNA encoding wild type mTOR and the glutamate induced current (I_glu), a measure of glutamate transport, determined by dual electrode voltage clamp. Moreover, EAAT3 protein abundance was determined utilizing chemiluminescence. As a result, I_glu was observed in Xenopus oocytes expressing EAAT3 but not in water injected oocytes. Coexpression of mTOR significantly increased I_glu, an effect reversed by rapamycin (100 nM). mTOR coexpression increased EAAT3 protein abundance in the cell membrane. The decay of I_glu following inhibition of carrier insertion with brefeldin A in oocytes coexpressing EAAT3 + mTOR was similar in the presence and absence of rapamycin (100 nM). In conclusion, mTOR is a novel powerful regulator of glutamate transporters and may thus contribute to protection against neuroexcitotoxicity.