FIGURE 8.

Nucleotide binding is required for efficient glutamate-induced synaptic translocation of GFP-CaMKII in hippocampal neurons. A, GFP-CaMKII localization was monitored in live rat hippocampal neurons (DIV 13) before and during a 10 min glutamate/glycine (100 μm/10 μm) stimulus. Translocation to synaptic sites was seen for both wild type and T286A CaMKII within 4 min of stimulation, whereas K42M did not display this pattern of localization even after 10 min of stimulation. Shown are sample projection images created from deconvolved z-stacks of the same neuron at different time points. B, translocation in time lapse images was quantified (for 20 neurons in six independent experiments for each condition). CaMKII wild type and the T286A mutant showed the same translocation, while K42M translocated significantly less than T286A (and wild type) as indicated (*, p < 0.05; **, p < 0.01; in Bonferroni post-hoc analysis after two-way ANOVA).