Figure 3. Activity-dependent... | Rockefeller University Press

Figure 3.

$Figure 3. Activity-dependent translocation of αCaMKII to microtubules. (Ai) Time series of FRAP of mGFP-αCaMKII during a 0Mg2+/Gly stimulation in a dendrite, either where CaMKII has concentrated (D+, white brackets) or not (D−, red brackets). White circles indicate regions of photobleaching. Bar, 5 µm. (Aii) FRAP curves (bleaching at t = 3 s) and (Aiii) mobile fractions of CaMKII during and after a 0Mg2+/Gly stimulation in corresponding regions. (B) Dendrite from a neuron expressing mGFP-αCaMKII stimulated with 0Mg2+/Gly for 5 min. Bar, 5 µm. (C–Di) Time-lapse imaging of mGFP-αCaMKII (C) or mCherry-αCaMKII and GFP-MAP2B (D) during a 1-min stimulation with KCl. Bars: (neurons) 10 µm; (inset) 2 µm. (Ei) Time series of FRAP of mGFP-αCaMKII before (top) and during (bottom) a 1-min KCl stimulation. White circles indicate regions of photobleaching. Bar, 3 µm. (Eii) FRAP curves and (Eiii) mobile fractions before and during the stimulation on neurons treated or not with 5 µM nocodazole. Arrows in B, C, and D point to synaptic (red) or microtubule-like sites (white) of CaMKII translocation. *, P < 0.05 unpaired t test; ‡, P < 0.05 paired t test. (Fi) Neuron expressing mCherry-αCaMKII and GFP (top) or GFP-MAP2B (bottom). White arrows point to detectable fibers in CaMKII signal. Bar, 10 µm. (Fii) Quantification of mCherry-αCaMKII microtubular localization in high Mg2+/low Ca2+ solution when cotransfected with GFP or GFP-MAP2B. n = 49–51 neurons per condition. *, P < 0.05 unpaired t test. See also Figs. S2, S3, S4 and S5 A, and Video 5 and 6.$

Activity-dependent translocation of αCaMKII to microtubules. (Ai) Time series of FRAP of mGFP-αCaMKII during a 0Mg²⁺/Gly stimulation in a dendrite, either where CaMKII has concentrated (D+, white brackets) or not (D−, red brackets). White circles indicate regions of photobleaching. Bar, 5 µm. (Aii) FRAP curves (bleaching at t = 3 s) and (Aiii) mobile fractions of CaMKII during and after a 0Mg²⁺/Gly stimulation in corresponding regions. (B) Dendrite from a neuron expressing mGFP-αCaMKII stimulated with 0Mg²⁺/Gly for 5 min. Bar, 5 µm. (C–Di) Time-lapse imaging of mGFP-αCaMKII (C) or mCherry-αCaMKII and GFP-MAP2B (D) during a 1-min stimulation with KCl. Bars: (neurons) 10 µm; (inset) 2 µm. (Ei) Time series of FRAP of mGFP-αCaMKII before (top) and during (bottom) a 1-min KCl stimulation. White circles indicate regions of photobleaching. Bar, 3 µm. (Eii) FRAP curves and (Eiii) mobile fractions before and during the stimulation on neurons treated or not with 5 µM nocodazole. Arrows in B, C, and D point to synaptic (red) or microtubule-like sites (white) of CaMKII translocation. *, P < 0.05 unpaired t test; ‡, P < 0.05 paired t test. (Fi) Neuron expressing mCherry-αCaMKII and GFP (top) or GFP-MAP2B (bottom). White arrows point to detectable fibers in CaMKII signal. Bar, 10 µm. (Fii) Quantification of mCherry-αCaMKII microtubular localization in high Mg²⁺/low Ca²⁺ solution when cotransfected with GFP or GFP-MAP2B. n = 49–51 neurons per condition. *, P < 0.05 unpaired t test. See also Figs. S2, S3, S4 and S5 A, and Video 5 and 6.