Figure 7: pT305-CaMKII and memory consolidation in Lymnaea.

(a) Timeline for the behavioural experiment (conducted concurrently with the biochemical assays shown in b and c) is shown above the diagram. The classically conditioned and CaMKIINtide-treated animals (N=23) as well as the Naive animals (N=16) showed a significantly lower (asterisks) feeding response to the CS compared with the classically conditioned vehicle-treated animals (N=21). One-way ANOVA: P<0.0001. Tukey’s tests: CaMKIINtide and Naive versus Vehicle, both P<0.05, CaMKIINtide versus Naive, P>0.05. (b,c) The selective effects of CaMKIINtide on T305 phosphorylation of CaMKII. Examples of western blot bands are shown above the graphs (full-length blots in Supplementary Fig. 7d,e). The means (±s.e.m.) of pT305-CaMKII (b) or pT286-CaMKII (c) over CaMKII levels are shown (relative integrated density). Total CaMKII levels were not significantly different among the groups. The values thus calculated for each CaMKIINtide-treated (b, N=7; c, N=7) and vehicle-treated sample (b, N=6; c, N=6) were normalized to the mean of the density values obtained from the naive samples (b, N=14; c, N=12), providing a baseline of 1 (dashed line). Vehicle treatment 6 h after paired training leaves the significantly raised level of pT305-CaMKII (see Fig. 6a) unaffected, whereas CaMKIINtide treatment significantly decreases it (b). Asterisks indicate significant differences both compared to the Naive baseline and between the two paired groups. One-sample t-tests: CaMKIINtide and Vehicle versus baseline, both P<0.03. Two-sample t-test: CaMKIINtide versus Vehicle, P<0.002. T286 phosphorylation remains unaffected by treatment with either vehicle or CaMKIINtide (c). One-sample t-tests: CaMKIINtide and Vehicle versus baseline, P=0.70 and 0.69, respectively. Two-sample t-tests: CaMKIINtide versus Vehicle, P=0.90. These experiments were replicated twice.