In situations of hypoxia, glutamate excitotoxicity induces neuronal death. The release of extracellular adenosine is also triggered and is accompanied by an increase of the stress mediator, corticotrophin-releasing factor (CRF). Adenosine A(2A) receptors contribute to glutamate excitoxicity and their blockade is effective in stress-induced neuronal deficits, but the involvement of CRF on this effect was never explored. We now evaluated the interaction between A(2A) and CRF receptors (CRFR) function, upon glutamate insult. Primary rat cortical neuronal cultures (9 days in vitro) expressing both CRF(1) R and CRF(2) R were challenged with glutamate (20-1000μM, 24 hours). We found CRF(1) R colocalized with neuronal markers and CRF(2) R present in both neuronal and glial cells. The effects of the CRF and A(2A) receptors ligands on cell viability were measured using propidium iodide and Syto-13 fluorescence staining. Glutamate decreased cell viability in a concentration-dependent manner. Urocortin (10pM), a CRF receptors agonist, increased cell survival in the presence of glutamate. This neuroprotective effect was abolished by blocking either CRF(1) R or CRF(2) R with antalarmin (10nM) or anti-Sauvagine-30 (10nM), respectively. The blockade of A(2A) receptors with a selective antagonist SCH 58261 (50nM) improved cell viability against the glutamate insult. This effect was dependent on CRF(2) R, but not on CRF(1) R activation. Overall these data show a protective role of CRF in cortical neurons, against glutamate-induced death. The neuroprotection achieved by A(2A) receptors blockade requires CRF(2) R activation. This interaction between the adenosine and CRF receptors can explain the beneficial effects of using A(2A) receptor antagonists against stress-induced noxious effects. © 2012 International Society for Neurochemistry, J. Neurochem. (2012) 10.1111/jnc.12050.