Fig. 5

Os-pep restored synaptic functions in mice and in vitro with AD-mimicking pathological conditions and Adipo^−/− mice. (a-c) Immunoblotting and quantification of Synap, SNAP-23, SAP102 and PSD95 levels in the AβO, APP/PS1 and Adipo^−/− mice respectively. The data are expressed as the means ± SEM for the indicated proteins in vivo (n = 8 mice/group) and the number of independent experiments = 3. Significance = ***p < 0.001; ###p < 0.001; One-way ANOVA followed by Turkey’s post hoc test. ns, indicated that there is no significant difference among the control, sham and Os-pep treated groups (d) Immunoblotting and quantification of the Synap and PSD95 levels in APPswe/ind-overexpressed SH-SY5Y cells transfected with the AdipoR1 siRNA and treated with Os-pep (10 μM). The data are expressed as the means ± SEM for the indicated proteins in vitro (n = 5/group), and the number of independent experiments = 3. Significance = ***p < 0.001; ###p < 0.001; Two-way ANOVA followed by Bonferroni’s multiple comparisons test (e) Immunoblotting and quantification of the Synap and PSD95 levels in APPswe/ind-overexpressed SH-SY5YIR cells exposed to a high insulin (1 μM) concentration, a low insulin concentration (10 nM), Os-pep (10 μM) and Comp. C (10 μM) and transfected with the AdipoR1 siRNA. The data are expressed as the means ± SEM for the indicated proteins in vitro (n = 5/group), and the number of independent experiments = 3. Significance = ***p < 0.01, **p < 0.01; ###p < 0.001; Two-way ANOVA followed by Bonferroni’s multiple comparisons test. (f) Os-pep regulated dendritic complexity and spine density in the Adipo^−/− mice. (f, panel (a)) Indicated images were the example of the reconstructed hippocampal CA1 region of pyramidal neurons in the WT (Veh), Adipo^−/− and Os-pep treated Adipo^−/− (Os-pep). (f, panel (b)) Representative histogram indicated total dendritic length (sum of basal and apical dendrite length) in the hippocampal CA1 region of the mice. (f, panel (c)) Representative histogram indicated dendritic complexity via using the sholl analysis of reconstructed pyramidal neurons. (f, panel (d)) Representative images of digitalized hippocampal CA1 pyramidal dendrites from the secondary branches. (f, panel (e)) Representative histograms indicate the dendritic spines density; including the total number of spines, the number of filopodia-like, thin, mushroom and stubby spines in the hippocampal CA1 region of brain mice. The data are shown as the mean ± SEM of 45 pyramidal neurons and 300 dendritic segments per groups and the number of independent experiments = 3. Significance = *p < 0.05; #p < 0.05; One-way ANOVA followed by Turkey’s post hoc test