Figure 8

Glutamate-induced the cleavage of α-fodrin in primary cortical cells. A: primary cortical cells isolated from 18 day old fetal rat brains were cultured for 7–8 days and then cells were treated with 1 mM glutamate for the indicated times. Western blot analysis showed that calpain-mediated 145 kDa BDP and caspase-3-mediated 120 kDa BDP could be detected in untreated control cells. Accumulation of 145 kDa BDP was evident at 0.5 hr earlier than that of 120 kDa BDP, with greater accumulation of both breakdown products occurring at 6 hrs. B: primary cortical cells were cultured for the indicated days and the cleavage of α-fodrin was detected by Western blot. Tissue sample only contained the intact 240 kDa α-fodrin protein but no cleavage products. Cleavage of the 240 kDa α-fodrin protein into 145 kDa BDP began at day 0 of culture (dissociated cells prior to seeding on plates) and the 120 kDa BDP of α-fodrin was only found after 7 days of culture. Each experiment was repeated 3 times and a representative example of the Western blot is shown.