Cortical cultures of rat brain neurons were exposed to ethanol (100 mM) for 4 days in order to examine whether the pharmacological characteristics of N-methyl-D-aspartate (NMDA) receptors expressed by these neurons were altered by this treatment. In fura-2 loaded control neurons, NMDA (plus 10 microM glycine) stimulated a dose-dependent increase in intracellular calcium concentrations with an estimated EC50 value of 6.8 microM. NMDA-stimulated increases in intracellular calcium reached a plateau at approximately 30 microM with no further increases observed at 100 microM. The EC50 value for NMDA in ethanol-exposed neurons was reduced to 1.8 microM with no alteration in the maximal response. Similarly, the EC50 value for glycine (tested with 100 microM NMDA) was reduced from 2.3 microM in control cultures to 0.67 microM in ethanol-treated cultures. Ifenprodil inhibited NMDA-stimulated increases in intracellular calcium in control cultures only at concentrations of 3 microM and above, with 100 microM producing approximately a 58% inhibition. In ethanol-treated cultures, 0.3 microM ifenprodil inhibited the NMDA response by approximately 60% with 100 microM ifenprodil producing a 72% inhibition. Over the concentration range of ifenprodil tested, half-maximal inhibition occurred at 1.4 microM and 0.18 microM, respectively, for control and ethanol-treated neurons. Although chronic ethanol treatment appeared to alter the sensitivity of neurons to NMDA agonists and antagonists, the inhibitory effects of 50 mM ethanol on NMDA-stimulated increases in intracellular calcium were not different between control (28% inhibition) and ethanol-treated neurons (27% inhibition). Finally, the changes in NMDA receptor sensitivity observed in ethanol-treated neurons were accompanied by an enhanced sensitivity to the neurotoxic effects of NMDA as measured by propidium iodide staining. These results suggest that chronic exposure of neurons to ethanol may result in an altered expression of agonist-sensitive/ifenprodil selective NMDA receptor subunits.