Figure 5

Monitoring the molecular dynamics of αCaMKII in living cortical neurons. (A, C) Typical images of the interaction of YFP-αCaMKII with CFP-CaM (A) and the conformational change of YFP-αCaMKII-CFP (C) in the cytoplasm (upper panels) and dendrite (lower panels) before and after ionomycin application (0.5 μM) for 10 min. Monochrome images show YFP fluorescence. (B, D) Left graphs show relative emission ratio and right graphs show FRET efficiencies before (white bars) and after (black bars) ionomycin application. Upper graphs show the results of cytoplasm. Lower graphs show the results of dendrites. (E, F) Changes in relative emission ratio following the application of ionomycin or KCl (open circle, ionomycin; filled circle, KCl). Black bar shows the period of ionomycin or KCl application. 9–10 cells were analyzed in each group. (G, H) Effect of short or long-term activation of NMDA receptor by the application of NMDA (25 μM) with or without pre-treatment of AP5 (25 μM) (open circle, long-term; gray circle, short-term; filled circles, long-term with pre-treatment of AP5). Black bar and white bar show the period of NMDA application. 7–9 cells were analyzed in each group.