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. 2020 Feb 4;10:1777. doi: 10.1038/s41598-020-58610-6

Figure 1.

Figure 1

PKC regulates Rac1 activation during sLTP. (a) Schematic of small GTPase FRET sensors. (b) 2pFLIM images of Rac1 activation in WT slices at indicated time points. Arrowhead represents point of uncaging. Warmer colors indicate higher binding fraction of sensor and higher Rac1 activity. Scale bar, 1 μm. (c) Time courses and quantification of transient (1–3 min) and sustained (10–25 min) spine volume change induced by glutamate uncaging in neurons expressing GFP or in neurons from PKCα WT and KO littermates expressing Rac1 sensor. (d) Time courses and quantification of transient (1.5–3.5 min), sustained (10–25 min) and basal (−8–0 min) Rac1 activation in stimulated spines from WT and PKCα KO littermates. (e) 2pFLIM images of Rac1 activation in PKCα KO slices at indicated time points. Arrowhead represents point of uncaging. Spreading of Rac1 activation in the dendrite were measured in the regions 0 μm from the stimulated spine (red), 1 μm (orange), 2 μm (yellow), 3 μm (brown) and 4 μm (purple). Scale bar, 1 μm. (f) Spatial profile and quantification of spreading Rac1 activation along the dendrite at indicated times and distances from the stimulated spine in WT and PKCα KO littermates. Data are mean ± s.e.m. Grey shading indicates time of uncaging. *P < 0.05, **P < 0.01 two-tailed t-test (c,d) and two-way ANOVA with Sidak’s mutiple comparisons test (f). n (neurons/spines) = 18/22 WT, 19/22 PKCα KO and 5/11 GFP.