Differential potentiation by spermidine of abilities of a variety of displacers for [3H]MK-801 binding in hippocampal synaptic membranes.

Binding of 3H-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imi ne (MK-801) to an ion channel associated with the N-methyl-D-aspartate (NMDA)-sensitive subtype of brain excitatory amino acid receptors was studied in Triton-treated preparations of synaptic membranes of rat brain. The initial association rate of the binding measured at 30 min after onset of incubation was markedly potentiated by the addition of either L-glutamic acid (Glu) alone or both Glu and glycine (Gly) in a concentration-dependent manner at 10 nM to 0.1 mM. Potentiation occurred to a significantly greater extent in the hippocampus and cerebral cortex than in the cerebellum. In the presence of both Glu and Gly, the endogenous polyamine spermidine (SPD) further potentiated binding in hippocampal and cortical membranes at concentrations above 10 microM without significantly affecting that in cerebellar membranes. The binding of [3H]MK-801 was slowly equilibrated in 16 h. When examined in hippocampal synaptic membranes, the binding at equilibrium was markedly displaced by numerous noncompetitive antagonists for the NMDA receptor. The addition of SPD markedly enhanced potencies of those displacers having a high affinity to [3H]MK-801 binding sites, without affecting other displacers having a low affinity. These results suggest that SPD promotes transition of sites responsible for mediating NMDA responses within the channel to a state with higher affinity for noncompetitive blockers.