This study utilized a grease-gap preparation to investigate the effects of polyamines on responses of CA1 hippocampal pyramidal cells to N-methyl-D-aspartate (NMDA) and on the block of the NMDA channel by Mg2+. In the absence of added Mg2+, 1,10-diaminodecane (0.1-1 mM) non-competitively antagonized NMDA-evoked depolarizations. Its antagonism slowly progressed to a stable value, was not use-dependent and did not reverse completely upon washout. Similar results were obtained with 100 microM spermine and 1 mM diethylenetriamine. Addition of 1 mM Mg2+ to the superfusion medium greatly reduced these effects. Conversely, the polyamines attenuated the blocking action of Mg2+. Postnatal treatment with alpha-difluoromethylornithine reduced the total polyamine content of area CA1 in 10- to 15-day-old rats almost to the adult level (although spermine content was unaffected). Mg2+ less potently antagonized NMDA-evoked depolarizations in slices from 10- to 15-day-old rats than in slices from adult rats, and this difference was unaffected by the alpha-difluoromethylornithine treatment. These results suggest (1) that there are rapid and slow components to the antagonism of NMDA-evoked depolarizations by polyamines, both of which may involve permeation of the polyamine into or through the NMDA channel: (2) that polyamine release in brain could modulate the Mg2+ sensitivity of responses to NMDA; and (3) that changes in the total content of endogenous polyamine do not explain developmental differences in the sensitivity of NMDA-evoked depolarizations to Mg2+.