Fig. 2
Spatial memory and gene expression depend on NMDAR- and CaV1-mediated signaling via γCaMKII/CaM translocation. a Significant impairment in the ability of γCaMKII exc-KO mice during the acquisition phase of the MWM test (n = 12 mice for each group). No difference was detected between behavior of exc-KO and KO animals (p > 0.16, one-way ANOVA) or between the two cohorts of WT mice (p > 0.15, one-way ANOVA). b Representative images show nuclear CaM and c-Fos expression in the pyramidal layer of CA1 (denoted by white lines) of WT mice at 1 h after MWM training, which are absent in γCaMKII exc-KO mice. Scale bar, 20 μm. c Percentage of c-Fos-positive cells plotted against nuclear CaM level in CA1 of WT or exc-KO mice, 1 h after MWM training (n = 5 mice for each group, p = 0.04 for c-Fos and p = 0.006 for CaM). Vertical and horizontal dotted lines represent CaM and c-Fos nuclear values in naive (untrained) mice. d–k Cultured cortical neurons were either “mock-stimulated” (basal) or stimulated with 25 μM NMDA and 5 μM glycine, without (control, ctl) or with γCaMKII shRNA knockdown (d–g), or without (control, labeled ‘-‘) or with nimodipine, KN93, PD98059, or APV. Following fixation, cells were stained for CaM (d, h), pCaMKIV (e, i), pCREB (f, j), or c-Fos (g, k). All plots are the average of two experiments with >25 cells per condition (one-way ANOVA followed by Student’s t-test). l Schematic diagram of CaV1- and NMDA receptor-driven, γCaMKII-mediated cytoplasm-to-nucleus signaling19,27,44. Colors refer to signaling intermediates assessed in d–k. For more details on alternative pathways, see Supplementary Fig. 5m. Statistical analysis was performed with one-way ANOVA followed by Holm–Sidak post hoc test unless otherwise noted. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. Error bars represent SEM
