rs γ2  (Bioss)

Journal: Translational Psychiatry

Article Title: Caspase-1 affects chronic restraint stress-induced depression-like behaviors by modifying GABAergic dysfunction in the hippocampus

doi: 10.1038/s41398-023-02527-x

Figure Lengend Snippet: A Hippocampal GAD65, GAD67 and VGAT mRNA were determined by qPCR after CRS ( n = 6 mice/group, Student’s test). B Correlation of the levels of GAD67 mRNA with sucrose preference after CRS (Pearson correlation). C Representative immunoblots and quantification of GAD67 levels from Control and CRS mice ( n = 6-7 mice/group, Student’s test). D CRS reduced the levels of GABA in the hippocampus ( n = 9-10 mice/group, Student’s test). E Representative whole-cell voltage-clamp traces of GABA A Rs-mediated mIPSCs in the pyramidal neurons of hippocampus from Control and CRS mice. Scale bars: 25 pA, 1 s. F , G Amplitude and frequency of GABA A Rs-mediated mIPSCs from Control and CRS mice ( n = 8-9 cells from 3-4 mice/group, Student’s test). H Representative immunoblots and quantification of the surface levels of GABA A Rs γ2 subunit (sGABA A Rs γ2) and total levels of GABA A Rs γ2 subunit (tGABA A Rs γ2) in the hippocampus from Control and CRS mice ( n = 5 mice/group, Student’s test). I Representative immunoblots and quantification of parvalbumin (PV) levels from Control and CRS mice ( n = 5 mice/group, Student’s test). J Immunohistochemistry against PV-positive GABAergic interneurons in the hippocampus. Scale bars: 200 μm. All data are shown as means ± SEM. * p < 0.05 and ** p < 0.01.

Article Snippet: After blocking with 5% BSA for 1 h at room temperature, the membranes were incubated overnight at 4 °C with the primary antibodies: β-actin (1:3000, abs137975, absin, China); GAD67 (1:500, MAB5406, Merk Millipore, USA); caspase-1 (1:1000, 06-503-I, Merk Millipore, USA); GABA A Rs γ2 (1:400, bs-4112R, Bioss, China); GAPDH (1:3000, abs132004, absin, China); parvalbumin (PV) (1:200; bs-1299R, Bioss, China).

Techniques: Western Blot, Control, Immunohistochemistry

Journal: Translational Psychiatry

Article Title: Caspase-1 affects chronic restraint stress-induced depression-like behaviors by modifying GABAergic dysfunction in the hippocampus

doi: 10.1038/s41398-023-02527-x

Figure Lengend Snippet: A A schematic diagram depicting the experimental procedure. D, day. B Body weight throughout the CRS treatment ( n = 9-10 mice/group, repeated-measures ANOVA: CRS, F (7,238) = 2.301, p < 0.05; Genotype, F (7,238) = 1.345, p > 0.05; Interaction, F (7,238) = 2.101, p < 0.05). C–E Gene ablation of caspase-1 prevented CRS-induced anhedonia in SPT (two-way ANOVA: CRS, F (1,34) = 15.934, p < 0.001; Genotype, F (1,34) = 7.217, p < 0.05; Interaction, F (1,34) = 5.783, p < 0.05), and increased immobility time in TST (two-way ANOVA: CRS, F (1,34) = 4.154, p < 0.05; Genotype, F (1,34) = 7.372, p < 0.05; Interaction, F (1,34) = 4.441, p < 0.05) and FST (two-way ANOVA: CRS, F (1,34) = 4.995, p < 0.05; Genotype, F (1,34) = 10.452, p < 0.001; Interaction, F (1,34) = 5.490, p < 0.05) ( n = 9-10 mice/group, two-way ANOVA, Bonferroni’s test). F Serum corticosterone (CRS, F (1,33) = 32.745, p < 0.001; Genotype, F (1,33) = 0.904, p > 0.05; Interaction, F (1,33) = 0.02, p > 0.05) and IL-1β (CRS, F (1,33) = 9.160, p < 0.01; Genotype, F (1,33) = 8.120, p < 0.01; Interaction, F (1,33) = 7.058, p < 0.05) from WT, WT-CRS, Caspase-1 −/− and Caspase-1 −/− -CRS groups ( n = 9-10 mice/grou p , two-way ANOVA). G Caspase-1 deletion blocked CRS-induced increase of IL-1β mRNA in the hippocam p us ( n = 4 mice/group, two-way ANOVA: CRS, F (1,12) = 61.957, p < 0.001; Genotype, F (1,12) = 37.686, p < 0.001; Interaction, F (1,12) = 58.990, p < 0.001). All data are shown as means ± SEM. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: After blocking with 5% BSA for 1 h at room temperature, the membranes were incubated overnight at 4 °C with the primary antibodies: β-actin (1:3000, abs137975, absin, China); GAD67 (1:500, MAB5406, Merk Millipore, USA); caspase-1 (1:1000, 06-503-I, Merk Millipore, USA); GABA A Rs γ2 (1:400, bs-4112R, Bioss, China); GAPDH (1:3000, abs132004, absin, China); parvalbumin (PV) (1:200; bs-1299R, Bioss, China).

Techniques:

Journal: Translational Psychiatry

Article Title: Caspase-1 affects chronic restraint stress-induced depression-like behaviors by modifying GABAergic dysfunction in the hippocampus

doi: 10.1038/s41398-023-02527-x

Figure Lengend Snippet: A Hippocampal GAD65 (two-way ANOVA: CRS, F (1,12) = 0.671, p > 0.05; Genotype, F (1,12) = 0.735, p > 0.05; Interaction, F (1,12) = 0.392, p > 0.05), GAD67 (two-way ANOVA: CRS, F (1,12) = 6.604, p < 0.05; Genotype, F (1,12) = 7.439, p < 0.05; Interaction, F (1,12) = 10.025, p < 0.01) and VGAT (two-way ANOVA: CRS, F (1,12) = 0.277, p > 0.05; Genotype, Genotype, F (1,12) = 1.501, p > 0.05; Interaction, F (1,12) = 0.193, p > 0.05) mRNA were determined by qPCR from WT, WT-CRS, Caspase-1 −/− and Caspase-1 −/− -CRS groups ( n = 4 mice/group, two-way ANOVA, Bonferroni’s test). B Representative immunoblots and quantification of GAD67 levels from different groups ( n = 5 mice/group, two-way ANOVA: CRS, F (1,16) = 6.024, p < 0.05; Genotype, F (1,16) = 9.735, p < 0.01; Interaction, F (1,16) = 15.582, p < 0.01). C Caspase-1 deficiency prevented CRS-induced reduction of GABA in the hippocampus ( n = 9-10 mice/group, two-way ANOVA: CRS, F (1,34) = 5.861, p < 0.05; Genotype, F (1,34) = 6.634, p < 0.05; Interaction, F (1,34) = 6.466, p < 0.05). D Representative whole-cell voltage-clamp traces of GABA A Rs-mediated mIPSCs in the pyramidal neurons of hippocampus from different groups. Scale bars: 25 pA, 1 s. E , F Amplitude (two-way ANOVA: CRS, F (1,28) = 16.962, p < 0.001; Genotype, F (1,28) = 13.275, p < 0.01; Interaction, F (1,28) = 15.973, p < 0.001) and frequency (two-way ANOVA: CRS, F (1,28) = 1.887, p > 0.05; Genotype, F (1,28) = 0.482, p > 0.05; Interaction, F (1,28) = 6.481, p < 0.05) of GABA A Rs-mediated mIPSCs from different groups (n = 8 cells from 3-4 mice/group, two-way ANOVA, Bonferroni’s test). G Representative immunoblots and quantification of sGABA A Rs γ2 (CRS, F (1,16) = 7.821, p < 0.05; Genotype, F (1,16) = 6.178, p < 0.05; Interaction, F (1,16) = 5.994, p < 0.05) and tGABA A Rs γ2 (CRS, F (1,16) = 0.124, p > 0.05; Genotype, F (1,16) = 0.140, p > 0.05; Interaction, F (1,16) = 0.143, p > 0.05) levels in the hip p ocampus from different groups ( n = 5 mice/group, two-way ANOVA). All data are shown as means ± SEM. * p < 0.05 and ** p < 0.01.

Article Snippet: After blocking with 5% BSA for 1 h at room temperature, the membranes were incubated overnight at 4 °C with the primary antibodies: β-actin (1:3000, abs137975, absin, China); GAD67 (1:500, MAB5406, Merk Millipore, USA); caspase-1 (1:1000, 06-503-I, Merk Millipore, USA); GABA A Rs γ2 (1:400, bs-4112R, Bioss, China); GAPDH (1:3000, abs132004, absin, China); parvalbumin (PV) (1:200; bs-1299R, Bioss, China).

Techniques: Western Blot

Journal: Translational Psychiatry

Article Title: Caspase-1 affects chronic restraint stress-induced depression-like behaviors by modifying GABAergic dysfunction in the hippocampus

doi: 10.1038/s41398-023-02527-x

Figure Lengend Snippet: A Hippocampal GAD67 was determined by qPCR from AAV-GFP and AAV-Casp1-injected mice after STVS ( n = 6 mice/group, Student’s test). B Correlation of hippocampal GAD67 mRNA levels with sucrose preference after STVS (Pearson correlation). C Representative immunoblots and quantification of GAD67 protein expression in the hippocampus ( n = 6 mice/group, Student’s test). D Correlation of hippocampal GAD67 protein expression with sucrose preference after STVS (Pearson correlation). E GABA levels in the hippocampus after STVS ( n = 9 mice/group, Student’s test). F Representative whole-cell voltage-clamp traces of GABA A Rs-mediated mIPSCs in the pyramidal neurons of hippocampus from AAV-GFP and AAV-Casp1-injected mice after STVS. Scale bars: 25 pA, 1 s. G , H Amplitude and frequency of GABA A Rs-mediated mIPSCs ( n = 8 cells from 3-4 mice/group, Student’s test). I Representative immunoblots and quantification of sGABA A Rs γ2 and tGABA A Rs γ2 levels in the hippocampus ( n = 5 mice/group, Student’s test). All data are shown as means ± SEM. * p < 0.05 and ** p < 0.01.

Article Snippet: After blocking with 5% BSA for 1 h at room temperature, the membranes were incubated overnight at 4 °C with the primary antibodies: β-actin (1:3000, abs137975, absin, China); GAD67 (1:500, MAB5406, Merk Millipore, USA); caspase-1 (1:1000, 06-503-I, Merk Millipore, USA); GABA A Rs γ2 (1:400, bs-4112R, Bioss, China); GAPDH (1:3000, abs132004, absin, China); parvalbumin (PV) (1:200; bs-1299R, Bioss, China).

Techniques: Injection, Western Blot, Expressing