Fig. 3

CaMKIIα-CaM association in response to a single glutamate uncaging pulse. a Representative fluorescence lifetime images of CaMKIIα-CaM association in response to a single glutamate uncaging pulse. Warmer colors indicate lower fluorescence lifetime, corresponding to a higher binding fraction of mCherry-CaM to mEGFP-CaMKIIα. Scale bar, 1 µm. b Time course of CaMKIIα-CaM association in a stimulated spine (black) and nearby dendritic (blue). Inset is an expanded view of the rising phase. Black squares denote uncaging pulses. Analyzed from images in a. c Averaged changes in CaMKIIα-CaM association in spines and nearby dendrite (n = 28 spines/4 neurons). The orange curve indicates the decay of binding fraction change obtained by curve fitting of an exponential function: B(t) = B₀ exp(–t/τ), where B₀ is the initial binding fraction change, τ is the dissociation time constant. The time constant is obtained as τ = 2.9 ± 0.3 s. d Comparison of CaMKIIα-CaM association in response to a single pulse (c) and to a train of glutamate uncaging (Fig. 2d, e). e CaMKIIα conformation change measured with Green-Camuiα in response to a single pulse and a train of glutamate uncaging. Data from our previous publication¹⁰. All data are shown in mean ± sem, and sem of time constants is obtained by bootstrapping