Journal: Neurochemical Research

Article Title: Morin Improves Cognitive Deficits in an in Vivo Model of Vascular Dementia by Modulating the N-methyl-D-aspartate Receptor Signaling Pathways

doi: 10.1007/s11064-026-04717-7

Figure Lengend Snippet: The impact of morin on the expression of synaptic plasticity-related proteins in the hippocampus of VaD rats. a Western blots; b the protein levels of SYP; c the protein levels of PSD-95; d the expression levels of SYP ; e the expression levels of PSD-95; f the levels of glutamate. Data are presented as mean ± SD. Statistical analysis was performed by one-way ANOVA with Tukey’s post-hoc test (data met assumptions of normality and homoscedasticity)/Kruskal-Wallis with Dunn’s test (data did not meet assumptions). * indicates a significant difference from the Sham group; # indicates a significant difference 2VO group; *# indicates a significant difference from both the Sham and 2VO groups, with a p-value of less than 0.05 considered statistically significant

Article Snippet: Primary antibodies, all diluted at 1:1000, were sourced from Santa Cruz Biotechnology and included the following: PSD-95 (sc-32290, Santa Cruz Biotechnology, Inc.), SYP (sc-17750, Santa Cruz Biotechnology, Inc.), and β-actin (sc-517582, Santa Cruz Biotechnology, Inc.).

Techniques: Expressing, Western Blot

Journal: Cell death & disease

Article Title: SAPAP3 regulates epileptic seizures involving GluN2A in post-synaptic densities.

doi: 10.1038/s41419-022-04876-9

Figure Lengend Snippet: Fig. 1 Expression of SAPAP3 in the temporal cortex of patients with TLE. A Triple-label immunofluorescence showed that SAPAP3 (green) and GFAP (blue) were not coexpressed in astrocytes, but SAPAP3 (green) and MAP2 (red) were coexpressed in the cortical neurons of patients. The arrows indicate positive cells. B Representative western blots showing the expression of SAPAP3 in patients with TLE and controls. C The comparison of the mean OD value indicates that SAPAP3 expression in the TLE group (n = 20) is significantly higher than that in the control group (n = 10). Data are presented as means ± SEM. ***p < 0.001.

Article Snippet: The target proteins were blotted with primary antibodies against SAPAP3 (1:200, Proteintech), GluN1 (1:1000, Abcam), GluN2A (1:1000, Millipore), GluN2B (1:1000, Millipore), and GAPDH (1:5000, Abcam) overnight at 4 °C.

Techniques: Expressing, Western Blot, Comparison, Control

Journal: Cell death & disease

Article Title: SAPAP3 regulates epileptic seizures involving GluN2A in post-synaptic densities.

doi: 10.1038/s41419-022-04876-9

Figure Lengend Snippet: Fig. 2 SAPAP3 expression in the hippocampus and cortex of the mouse model. A Triple-label immunofluorescence showed that SAPAP3 (green) and GFAP (blue) were not coexpressed in astrocytes, but SAPAP3 (green) and MAP2 (red) were mainly coexpressed within mouse hippocampal (upper panel) and cortical neurons (middle panel). Triple-label immunofluorescence showed that SAPAP3 (green) and VGLUT1 (blue) were not coexpressed, but SAPAP3 (green) and PSD95 (red) were coexpressed (lower panel). B Representative western blot images and quantification of SAPAP3 expression showed a significant increase in SAPAP3 expression in the pilocarpine-induced mouse epilepsy model compared with controls (left panel, cortex; right panel, hippocampus). C Representative western blot images and quantification of SAPAP3 expression showed a significant increase in SAPAP3 expression in the PTZ-induced mouse epilepsy model compared with the controls (left panel, cortex; right panel, hippocampus). Data are presented as means ± SEM, n = 12 per group. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: The target proteins were blotted with primary antibodies against SAPAP3 (1:200, Proteintech), GluN1 (1:1000, Abcam), GluN2A (1:1000, Millipore), GluN2B (1:1000, Millipore), and GAPDH (1:5000, Abcam) overnight at 4 °C.

Techniques: Expressing, Western Blot

Journal: Cell death & disease

Article Title: SAPAP3 regulates epileptic seizures involving GluN2A in post-synaptic densities.

doi: 10.1038/s41419-022-04876-9

Figure Lengend Snippet: Fig. 3 Seizure-modifying effects of SAPAP3 on the pilocarpine-induced model and PTZ-kindling model. A SAPAP3-sh treatment significantly increased the latency time to the first spontaneous seizures and B reduced the numbers of pilocarpine-induced spontaneous seizures in the chronic period. Recombinant SAPAP3 yielded the opposite results (A, B). Data are presented as means ± SEM, n = 8–10. *p < 0.05; **p < 0.01; ***p < 0.001. C SAPAP3-sh treatment significantly increased the latency time in the pentylenetetrazol-induced kindling model and E reduced the severity of pentylenetetrazol-induced seizures. Recombinant SAPAP3 yielded the opposite results (C, D). Data are presented as means ± SEM, n = 10. ***p < 0.001. F Representative EEG recording of pilocarpine-induced status epilepticus. G Typical EEG showing that mice with spontaneous recurrent seizures exhibited epileptic discharges.

Article Snippet: The target proteins were blotted with primary antibodies against SAPAP3 (1:200, Proteintech), GluN1 (1:1000, Abcam), GluN2A (1:1000, Millipore), GluN2B (1:1000, Millipore), and GAPDH (1:5000, Abcam) overnight at 4 °C.

Techniques: Recombinant

Journal: Cell death & disease

Article Title: SAPAP3 regulates epileptic seizures involving GluN2A in post-synaptic densities.

doi: 10.1038/s41419-022-04876-9

Figure Lengend Snippet: Fig. 4 Electrophysiological changes in hippocampal neurons in slices after SAPAP3 knockdown and overexpression. A, C, D Representative traces of APs, mEPSCs and mIPSCs in slices from mice treated with SAPAP3-sh-con, SAPAP3-sh, SAPAP3-con and recombinant SAPAP3 recorded in Mg2+-free ACSF. B Compared with the SAPAP3-sh-con group, the SAPAP3-sh LV significantly decreased the average frequency of APs. Compared with the SAPAP3-con group, the SAPAP3 overexpression LV significantly increased the average frequency of APs. SAPAP3-sh LV treatment significantly decreased the average frequency (E) and amplitude (F) compared with those of the control group and its representative cumulative fractions. SAPAP3 overexpression LV treatment yielded the opposite results (E, F). The data did not show a significant difference in the frequency (G) and amplitudes (H) of the mIPSCs among the groups. Data are presented as means ± SEM, n = 6 per group. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: The target proteins were blotted with primary antibodies against SAPAP3 (1:200, Proteintech), GluN1 (1:1000, Abcam), GluN2A (1:1000, Millipore), GluN2B (1:1000, Millipore), and GAPDH (1:5000, Abcam) overnight at 4 °C.

Techniques: Knockdown, Over Expression, Recombinant, Control

Journal: Cell death & disease

Article Title: SAPAP3 regulates epileptic seizures involving GluN2A in post-synaptic densities.

doi: 10.1038/s41419-022-04876-9

Figure Lengend Snippet: Fig. 5 Perturbations in the evoked EPSCs in response to changes in the SAPAP3 levels in the different groups. A Representative traces of evoked AMPAR- (−70 mV) and NMDAR-mediated (+40 mV) EPSCs; NMDA receptor-mediated responses were measured at 50 ms poststimulus. B Summary graph showing that the NMDA/AMPA ratio was lower in the SAPAP3-sh-treated mice than in the controls. SAPAP3 overexpression from the recombinant SAPAP3 LV yielded the opposite results. C The SAPAP3-sh and SAPAP3 overexpression LV affected AMPAR-mediated EPSCs, but the difference was not significant. D The SAPAP3-sh LV significantly decreased the average amplitude of NMDAR-mediated EPSCs compared with that of the control group; SAPAP3 overexpression yielded the opposite results. E No significant difference in the PPRs for AMPAR-mediated EPSCs was found among the groups. Data are presented as means ± SEM, n = 6 per group. *p < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: The target proteins were blotted with primary antibodies against SAPAP3 (1:200, Proteintech), GluN1 (1:1000, Abcam), GluN2A (1:1000, Millipore), GluN2B (1:1000, Millipore), and GAPDH (1:5000, Abcam) overnight at 4 °C.

Techniques: Over Expression, Recombinant, Control

Journal: Cell death & disease

Article Title: SAPAP3 regulates epileptic seizures involving GluN2A in post-synaptic densities.

doi: 10.1038/s41419-022-04876-9

Figure Lengend Snippet: Fig. 6 Expression of GluN1, GluN2A and GluN2B in total homogenates and the PSD after the behavioral evaluation. A–C Representative western blot of the GluN2A levels in total tissue homogenates and subcellular fractions (A). No significant differences in the total GluN2A levels were found among the groups (B). Compared with the control group, the SAPAP3-sh vector significantly decreased the synaptic/total GluN2A ratio. SAPAP3 overexpression significantly increased the synaptic/total GluN2A ratio (C). Representative western blots of the GluN2B (D) and GluN1 levels (G) in total tissue homogenates and subcellular fractions. No significant differences in the total GluN2B and GluN1 levels were found among the groups (E and H). The synaptic/total GluN2B and GluN1 ratios were not significantly different compared with those of the control group (F and I). GAPDH served as a loading control for the total homogenates. Data are presented as means ± SEM, n = 4 per group. *p < 0.05; ***p < 0.001.

Article Snippet: The target proteins were blotted with primary antibodies against SAPAP3 (1:200, Proteintech), GluN1 (1:1000, Abcam), GluN2A (1:1000, Millipore), GluN2B (1:1000, Millipore), and GAPDH (1:5000, Abcam) overnight at 4 °C.

Techniques: Expressing, Western Blot, Control, Plasmid Preparation, Over Expression

Journal: Cell death & disease

Article Title: SAPAP3 regulates epileptic seizures involving GluN2A in post-synaptic densities.

doi: 10.1038/s41419-022-04876-9

Figure Lengend Snippet: Fig. 7 Ultrastructural morphological changes in the PSD. A Representative electron micrographs show the distribution of asymmetric synapses at a magnification of 25,000. The arrow shows an asymmetric synapse. B As shown in the bar graph, no significant difference in the density of asymmetric synapses was found among the groups (/30 μm2) (n = 24 in each group). C The thickness of the PSDs in the SAPAP3-sh group (n = 132) was significantly decreased compared with that of the PSDs in the SAPAP3-sh-con group (n = 110). Representative images of the different groups are shown in D. Synaptic vesicle (sv), postsynaptic density (PSD), presynaptic membrane (prm), synaptic clearance (sc). Data are presented as means ± SEM, *p < 0.05.

Article Snippet: The target proteins were blotted with primary antibodies against SAPAP3 (1:200, Proteintech), GluN1 (1:1000, Abcam), GluN2A (1:1000, Millipore), GluN2B (1:1000, Millipore), and GAPDH (1:5000, Abcam) overnight at 4 °C.

Techniques: Membrane