Kinetics of glutamate (1 mM) induced Cyto c release from mitochondria into cytosol in primary cortical cells. Primary cortical cells isolated from 19 day old fetal rat brains were cultured for 7 days and then treated with 1 mM glutamate for the indicated times. Mitochondrial and cytosol fractions were isolated as described in "Methods". Protein levels of Cyto c in mitochondria and cytosol were determined by Western Blot analysis as described under Figure 6. Actin bands were also used to monitor the same blot to verify consistency of cytosolic protein loading. The content of Cyto c was measured by densitometric scanning of the film. The results are the mean (± SEM) of at least three experiments. *P < 0.05 compared with control.