Figure 3

Necrotic neurons enhance microglial-induced neurotoxicity mediated by NMDAR. (A) Neurotoxicity of MCM was assayed in cell cultures of CGN after 24 hours. MCM obtained from non-stimulated cells or after stimulation with necrotic neurons for 20 hours, MCM (Nec.), were added to a final concentration of 20% to neuronal cultures. Neuronal viability was analysed by PI incorporation after 24 hours. (B) Neuronal cell death was quantified after exposure to MCM of non-stimulated cells, MCM of microglial cells cultured in glutamine-free medium, MCM (- Glutamine), or cultured with a glutaminase inhibitor, MCM (+DON). (C) The NMDAR inhibitor, MK-801, was added to neuronal cultures at the same time as MCM and neuronal viability was assayed as described before. Results are shown as mean ± SD and are representative of at least two experiments. NS, non-significant. *** p < 0.001 and * p < 0.05.