Evoked BDNF release in Mecp2
−/y NG neurons. BDNF release evoked by patterned electrical field stimulation was compared in dissociate cultures of P35 wild-type and Mecp2
−/y NG neurons. Cultures were stimulated once every 20 s, with 2 s trains of biphasic pulses at 100 Hz, and assayed for released BDNF by ELISA in situ and for cellular BDNF content by standard ELISA. A, Wild-type and Mecp2
−/y neurons released the same absolute amount of BDNF during the period of stimulation (stimulated release = total release after stimulation − unstimulated release as measured in control cultures). However, as shown in B, cellular BDNF content was significantly lower in mutant neurons compared with wild-type cells, as in vivo. Therefore, as shown in C, mutant neurons released a significantly greater percentage of total BDNF content during stimulation than did wild-type cells. Each bar represents the mean ± SEM from at least three independent experiments. *p < 0.05 (ANOVA).