Abstract

Activation of N-methyl-d-aspartate-selective ionotropic glutamate receptors (NMDA receptors) requires two agonists, glutamate and glycine. These ligands are thought to bind to the NR2 and NR1 subunits, respectively, apparently ruling out the formation of functional homomeric receptors. However, NMDA-mediated currents are observed when the mammalian NR1 subunit is expressed alone inXenopus laevis oocytes. These currents have been generally ascribed to a functional association between NR1 and the endogenous glutamate receptor subunit XenU1. To determine whether such a functional association does in fact occur, we have isolated cDNAs for both XenU1 and XenU1a, a presumed nonallelic counterpart. We investigated whether the coexpression of either XenU1 or XenU1a with NR1 in either X. laevis oocytes and human embryonic kidney (HEK) 293 cells had any effect on the observed NMDA receptor responses. In oocytes, coinjection of XenU1 with NR1 did not increase the observed currents compared with injection of NR1 alone; similarly, in HEK 293 cells, coexpression of XenU1 and NR1 did not result in the formation of functional channels. We also found no pharmacological or biochemical evidence for interaction between the two subunits. We conclude, therefore, that XenU1 does not associate with the NR1 subunit and that an alternative explanation must be sought for the channels observed when NR1 is expressed alone in oocytes.