Journal: Brain Sciences

Article Title: TPPU Pre-Treatment Rescues Dendritic Spine Loss and Alleviates Depressive Behaviours during the Latent Period in the Lithium Chloride-Pilocarpine-Induced Status Epilepticus Rat Model

doi: 10.3390/brainsci11111465

Figure Lengend Snippet: ( A , B ) The ratios of NR1 and p-NR1 subunits of NMDA receptor was significantly decreased in the hippocampus of the SE+PEG400 group compared with the Control group, which was significantly increased in the SE+0.1TPPU group compared with the SE+PEG400 group after TPPU treatment. ( C , D ) The ratios of p-ERK1/2/ERK1/2 and p-CREB/CREB in the hippocampus were significantly increased in the SE+0.1TPPU group compared with the SE+PEG400 group. ( A–D, one-way ANOVA test, n = 4 in every group, * p < 0.05 ** p < 0.01). The figure on the left showed the bands of NR1, p-NR1, CREB, p-CREB, ERK1/2, p-ERK1/2, and β-actin in the Control, SE+PEG400, and the SE+0.1TPPU groups measured by the western-blot method.Hip, hippocampus; NR1, N-methy1-D-aspartate 1 receptor; p-NR1, phospho(S889)-N-methy1-D-aspartate 1 receptor; CREB, cyclic adenosine monophosphate-response element binding protein; p-CREB, phosphor (Ser133)-cyclic adenosine monophosphate-response element binding protein; ERK1/2, extracellular regulated kinase; p-ERK1/2, phospho (Thr202/Tyr204)-p44/42 extracellular regulated kinase.

Article Snippet: The procedures were as follows: (1) protein extracts were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to cellulose acetate membranes; (2) the membranes were blocked using goat serum and incubated with primary antibodies including rabbit anti-CREB (43 kDa, 1:1000, CST, Boston, MA, USA), rabbit anti-CREB-phospho Ser133 (43 kDa, 1:1000, CST, Boston, MA, USA), rabbit anti-ERK1/2 (43 kDa, 1:1000, CST, Boston, MA, USA), and rabbit anti-phospho-p44/42 ERK1/2 (Thr202/Tyr204) (42/44 kDa, 1:1000, CST), rabbit anti-NMDAR1 (120 kDa, 1:1000, CST, Boston, MA, USA), and rabbit anti-NMDAR1-phosphoS889 (120 kDa, 1:1000, CST, Boston, MA, USA) at 4 °C for 24 h; (3) the rabbit anti-β-actin primary antibody (40 kDa, 1:1000, Beyotime, Shanghai, China) was used as an internal reference; (4) after 24 h later, the membrane was incubated with the goat anti-rabbit IgG secondary antibody (1:1000, Beyotime, Shanghai, China) for 2 h at RT; (5) the Tanon Image software (version 4100, Shanghai, China) was used to analyse the bands of target proteins.

Techniques: Western Blot, Binding Assay