Fig. 5.

Nucleotide binding is required for extra-synaptic clustering of GFP-CaMKII in hippocampal neurons, as demonstrated by the nucleotide binding-deficient mutant K42M. Primary hippocampal neurons (14 days in vitro) were stimulated with glutamate/glycine (200 μM/10 μM) two days after transfection and imaged live on a climate controlled Zeiss Axiovert 200M setup at 32°C (z-stack of 24 images, 0.5 μm apart). Shown are projection images after deconvolution using SlideBook software. K42M significantly reduced formation of extra-synaptic clusters in the soma, but not clusters in the dendrites that likely correspond to synapses. T286A showed significant clustering even under basal conditions.