Figure 3
Regulation of spine dynamics by CaMKII in dissociated neurons. (a) Time-lapse imaging of spines in dissociated hippocampal neurons expressing RFP and treated with the CaMKII inhibitor KN93 or its inactive analogue KN92. Gain and loss of spines are marked by arrows. Bar, 5 µm. (b) Enhancement of the spine turnover rate in neurons treated with KN93. Data are presented as the mean ± SEM, (neurons at 19–20 DIV, n = 12 cells from 3 independent cultures), ***p < 0.001, **p < 0.01, t-test. (c) Time-lapse imaging of spines in dissociated hippocampal neurons expressing RFP along with GFP-tagged wild-type CaMKII or the kinase-dead mutant of CaMKII (K42R). Newly formed spines are marked by arrows. Bar, 5 µm. (d) Pseudocolour images of neurons expressing GFP-tagged wild-type CaMKIIα or the K42R mutant at similar levels. Bar, 5 µm. (e) Quantification of the expression level of GFP-tagged wild-type CaMKIIα or the K42R mutant. Data are presented as the mean ± SEM (neurons at 14–16 DIV, n = 11 cells from 3 independent cultures for wild-type CaMKIIa, n = 20 cells from 3 independent cultures for K42R CaMKIIα), n.s. p > 0.05, t-test. (f) Quantification of the increase in spine density from 14 to 16 DIV in neurons expressing GFP-tagged wild-type CaMKIIα or the K42R mutant. Data are presented as the mean ± SEM, (neurons at 14–16 DIV, n = 11 cells from 3 independent cultures for wild-type CaMKIIa, n = 20 cells from 3 independent cultures for K42R CaMKIIα), **p < 0.01, t-test.
