Glutamate excitotoxicity decreases viability of cultured hippocampal GABAergic neurons.

<p>GABAergic and glutamatergic neurons in the cultures (DIV7) were identified by immunocytochemistry, using antibodies against GABA (A, E) and VGLUT1+2 (A, D). The total number of cells present in the analysed fields was calculated based on the number of nuclei, stained with the fluorescent dye Hoechst 33342. Data are presented as mean±SEM of 5 independent preparations (A). Excitotoxic stimulation of hippocampal neurons was performed by incubation with 125 µM glutamate, for 20 min, in fresh Neurobasal medium containing B27 supplement, and the cells were further incubated in the original medium for 14 h. Cell death was assessed with the recombinant Luciferase chemoluminescence assay with CellTiterGlo (B), or by fluorescence microscopy using the fluorescence dye Hoechst 33342 (C). In the latter condition GABAergic cells were identified by immunocytochemistry, using an antibody against GABA. Data are presented as mean±SEM of 5 independent experiments. Statistical analysis was performed using Student's <i>t</i>-test. ***<i>p</i><0.001.