Previous studies have shown that the N-methyl-d-aspartate (NMDA) receptor is an important target for the actions of ethanol in the brain. N-methyl-d-aspartate receptors are glutamate-activated ion channels that are highly expressed in neurons. They are activated during periods of significant glutamatergic synaptic activity and are an important source of the signaling molecule calcium in the postsynaptic spine. Alterations in the function of NMDA receptors by drugs or disease are associated with deficits in motor, sensory and cognitive processes of the brain. Acutely, ethanol inhibits ion flow through NMDA receptors whereas sustained exposure to ethanol can induce compensatory changes in the density and localization of the receptor. Defining factors that govern the acute ethanol sensitivity of NMDA receptors is an important step in understanding how an individual responds to ethanol. In the present study, we investigated the effect of calcium-calmodulin dependent protein kinase II (CaMKII) on the ethanol sensitivity of recombinant NMDA receptors. Calcium-calmodulin dependent protein kinase II is a major constituent of the postsynaptic density and is critically involved in various forms of learning and memory. NMDA receptor subunits were transiently expressed in human embryonic kidney 293 cells along with CaMKII-alpha or CaMKII-beta tagged with the green fluorescent protein. Whole cell currents were elicited by brief exposures to glutamate and were measured using patch-clamp electrophysiology. Neither CaMKII-alpha or CaMKII-beta had any significant effect on the ethanol inhibition of NR1/2A or NR1/2B receptors. Ethanol inhibition was also unaltered by deletion of CaMKII binding domains in NR1 or NR2 subunits or by phospho-site mutants that mimic or occlude CaMKII phosphorylation. Chronic treatment of cortical neurons with ethanol had no significant effect on the expression of CaMKII-alpha or CaMKII-beta. The results of this study suggest that CaMKII is not involved in regulating the acute ethanol sensitivity of NMDA receptors.