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. Author manuscript; available in PMC: 2018 Jul 12.
Published in final edited form as: Neuron. 2017 Mar 9;93(6):1315–1324.e3. doi: 10.1016/j.neuron.2017.02.032

Figure 3. ERK activation and effect of AP5 during sLTP of single dendritic spines induced with 2-photon glutamate uncaging.

Figure 3

(A) Representative fluorescence lifetime images of ERK activation during glutamate uncaging. Stimulated spine was marked with the white arrowhead.

(B) ERK activation during sLTP along the stimulated dendrite as a function of the distance from the base of the stimulated spine (n = 12/4; spines/neurons) (stimulated spine, red circle; dendrites, black circle).

(C) Average time course of fluorescence lifetime change of EKARet-cyto in stimulated spines (red line), adjacent dendrites (0 μm, blue line), adjacent spines (green line) and dendritic segments 13 μm away from the stimulated spine (black line) during spine structural plasticity induced with 2-photon glutamate uncaging. Same data set as in B. The inset shows a closer view of the initial 6 min.

(D) Quantification of fluorescence lifetime change in (B) and (C) at indicated time points.

(E) Average time-course of fluorescence lifetime change of EKARet-cyto in the stimulated spines and the adjacent dendrite in the absence (n = 5/3) or presence (n = 11/3) of AP5.

(F) Averaged time course of volume change in the stimulated spines and adjacent dendrites in the absence (n = 5/3) or presence (n = 11/3) of AP5 correlated to (E). The glutamate uncaging was indicated as the black bar in (E) (F).

(G–I) Quantification of peak fluorescence lifetime change of EKARet-cyto (G) correlated to (E), spine volume change during the transient phase (6 min; H) and spine volume change during the sustained phase (average of 27–36 min; I) of sLTP in the stimulated spines and adjacent dendrites in the absence (n = 5/3; spines/neurons) or presence (n = 11/3; spines/neurons) of AP5 correlated to (F). Spine volume changes were measured as changes in fluorescence intensity of EKARet-cyto in the stimulated spines. All data were presented as mean ± S.E.M. (error bars). Statistical significance was tested with one-way ANOVA followed by Tukey’s test or t-test (ns P > 0.05, *** P < 0.001, **** P < 0.0001). Error bars indicate S.E.M.

See also Figures S2, S3