Journal: Frontiers in Neuroscience

Article Title: Long-term aerobic exercise improves learning memory capacity and effects on oxidative stress levels and Keap1/Nrf2/GPX4 pathway in the hippocampus of APP/PS1 mice

doi: 10.3389/fnins.2024.1505650

Figure Lengend Snippet: Aerobic exercise regulates Keap1/Nrf2/GPX4 pathway of the hippocampus in APP/AS1 mice. (A) Representative western blots of Keap1, Nrf2, HO-1, and GPX4 expression. (B–E) Quantification data of Keap1, Nrf2, HO-1, and GPX4 protein expression. n = 4, ∗ p < 0.05, compared with the control group; # p < 0.05, compared with the model group; & p < 0.05, compared with the S-te group. C, control group; M, model group; S-te, short-term exercise group; L-te, long-term exercise group.

Article Snippet: The protein sample was then electrophoresed in 12% SDS-polyacrylamide gels, transferred onto the PVDF membrane, and blocked using 5% skimmed milk for 1 h. The membrane was incubated with diluted (1:1000) primary antibodies against Keap-1 antibody (ab178846; Abcam), Nrf2 antibody 28,205-1-AP; Proteintech, HO-1 antibody (13129-1-AP; Proteintech, Shanghai, China), or GPX4 antibody (14104-1-AP; Proteintech, Shanghai, China) at 4°C overnight.

Techniques: Western Blot, Expressing, Control

Journal: Nature Communications

Article Title: Neuropeptide SP protects against colitis and linked anxiety-like behavior through the putative roles of gut microbiota and metabolite inositol

doi: 10.1038/s41467-025-67904-0

Figure Lengend Snippet: a , b Heat maps revealing differential gene expression in FMT-DSS vs FMT-CON, FMT-DSS + SP vs FMT-DSS groups. n = 4 mice/group. The pathway-related genes were selected (log2 fold change at least > 1, p < 0.05). c , d KEGG analysis of total-, up- and down-regulated genes in the FMT-DSS group compared with the FMT-CON group (Top 20). n = 4 mice/group. e , f KEGG analysis of total-, up- and down-regulated genes in the FMT-DSS + SP group vs the FMT-DSS group (Top 20). n = 4 mice/group. g Quantitative real-time PCR analysis of mRNA expressions of NF-κB signaling pathway genes ( p65 and IKBα ) in the hippocampus. n = 4 mice/group. The whiskers indicate the minimum and maximum values observed within the range of Q1 − 1.5 × IQR to Q3 + 1.5 × IQR. The box reveals the interquartile range (IQR) between the 25th (Q1) and 75th (Q3) percentiles, and the line inside the box represents the median (50th percentile). h The protein expression of p-p65 and p-IKBα in hippocampus tissue, as determined by western blotting. n = 3 independent experiments. i Quantitative real-t i me PCR analysis of mRNA expressions of GABA receptor and Ca 2+ signaling genes ( Gabra1 , Gabra3 , Gabrg2 , Gabrb2 and Camk2d ). n = 4 mice/group. The whiskers indicate the minimum and maximum values observed within the range of Q1 − 1.5 × IQR to Q3 + 1.5 × IQR. The box reveals the interquartile range (IQR) between the 25th (Q1) and 75th (Q3) percentiles, and the line inside the box represents the median (50th percentile). j Protein expressions of GABA A R, GABA B R, GAD65 and CaMKII were measured in hippocampus tissue by western blot. n = 3 independent experiments. k Representative immunofluorescence images of double-labeling for p-IKBα (red)/Iba1 (green) in hippocampus tissues. Scale bar = 50 μm. n = 3 independent experiments. I Representative immunofluorescence images of double-labeling for GABA A R (green)/GFAP (red) in hippocampus tissues. Scale bar = 50 μm. n = 3 independent experiments. m Double immunofluorescence staining for CaMKII (green)/GFAP (red) in hippocampus tissues and statistical analysis. Scale bar = 50 μm. n = 3 independent experiments. n Differential gene expressions were presented by the heatmap in microglia of three groups, and GSEA analysis of microglia from FMT-DSS + SP group vs the FMT-DSS group. n = 4 mice/group. o Heat maps of differential gene expression in astrocytes from three groups and GSEA analysis of astrocytes from FMT-DSS + SP group vs the FMT-DSS group. n = 4 mice/group. Data were presented as means ± SD. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001. Source data are provided as a file.

Article Snippet: Next, primary antibody against GFAP (1:1000, A8335, 1:200, NB100-53809, USA), GABA A Rα1 (1:1000, 12708-1-AP, Proteintech, China), GABA B R2 (1:1000, 27567-1-AP, Proteintech, China), CaMKII (1:1000, 12716, Cell Signaling Technology, USA), GAD65 (1:1000, ab239372, Abcam, USA), CD206 (1:1000, ab64693, Abcam, USA), iNOS (1:1000, 18985-1-AP, Proteintech, China), CD80 (1:1000, 66406-1, Proteintech, China), p-p65 (1:1000, A00284-1, Boster, China), p-IKBα (1:1000, WLH3930, Wanleibio, China), and β-actin (50201, 1:1000, Kemei Borui Science and Technology, China) were incubated with the membranes overnight at 4 °C.

Techniques: Gene Expression, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Immunofluorescence, Labeling, Double Immunofluorescence Staining

Journal: Nature Communications

Article Title: Neuropeptide SP protects against colitis and linked anxiety-like behavior through the putative roles of gut microbiota and metabolite inositol

doi: 10.1038/s41467-025-67904-0

Figure Lengend Snippet: a Diagram illustrating the mouse model of colitis employed in this study, and inositol treatments are indicated (Some schematic elements were created by Figdraw.com). b Daily body weight changes. n = 8 mice/group. c Diseases activity index. n = 6 mice/group. d Macroscopic pictures of colons. e H&E-stained colon sections. Scale bar = 100 μm. n = 5 independent experiments. f Immunofluorescence staining for E-cadherin (red)/DAPI (blue) in colon tissues. Scale bar = 100 μm. n = 4 independent experiments. g The mRNA level of inflammatory cytokines ( IL-1β , IL-6 and TNF-α ) in the colon. n = 4 mice/group. h , i Representative movement tracks in the open field test and related bar graphs (Distance traveled, distance of center region and speed). n = 6 mice/group. j , k Track plot of the elevated plus maze, and statistical analysis including percentage of time spent in the open arms and percentage of times entering the open arms. n = 6 mice/group. l Representative H&E-stained hippocampus sections of three groups. Scale bar = 100 μm. n = 6 independent experiments. m Photomicrographs of Nissl staining in the hippocampus. Scale bar = 100 μm. n = 6 independent experiments. n The mRNA expressions of downstream cytokines ( IL-6 , IL-1β , TNF-α and IL-10 ) in the hippocampus tissue. n = 5 mice/group. o Representative immunohistochemistry images of Iba-1 in hippocampus, Scale bar = 100 μm. n = 3 independent experiments. p The concentration of GABA in hippocampus tissue. n = 6 mice/group. q The mRNA expression of GABA A Rα1 in hippocampus. n = 5 mice/group. r The protein level of GABA B R detected by western blot. n = 3 independent experiments. s Double immunofluorescence staining for GFAP (red)/GAT1 (green) in the DG and CA1 regions of hippocampus tissue. Scale bar = 50 μm. n = 3 independent experiments. Data were presented as means ± SD. For body weight change, two-way repeated-measures ANOVA was performed and the rest of the statistics was analyzed using one-way ANOVA followed by Tukey’s multiple comparisons test. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001. Source data are provided as a file.

Article Snippet: Next, primary antibody against GFAP (1:1000, A8335, 1:200, NB100-53809, USA), GABA A Rα1 (1:1000, 12708-1-AP, Proteintech, China), GABA B R2 (1:1000, 27567-1-AP, Proteintech, China), CaMKII (1:1000, 12716, Cell Signaling Technology, USA), GAD65 (1:1000, ab239372, Abcam, USA), CD206 (1:1000, ab64693, Abcam, USA), iNOS (1:1000, 18985-1-AP, Proteintech, China), CD80 (1:1000, 66406-1, Proteintech, China), p-p65 (1:1000, A00284-1, Boster, China), p-IKBα (1:1000, WLH3930, Wanleibio, China), and β-actin (50201, 1:1000, Kemei Borui Science and Technology, China) were incubated with the membranes overnight at 4 °C.

Techniques: Activity Assay, Staining, Immunofluorescence, Immunohistochemistry, Concentration Assay, Expressing, Western Blot, Double Immunofluorescence Staining

Journal: Nature Communications

Article Title: Neuropeptide SP protects against colitis and linked anxiety-like behavior through the putative roles of gut microbiota and metabolite inositol

doi: 10.1038/s41467-025-67904-0

Figure Lengend Snippet: a Schematic illustration of experimental design (Some schematic elements were created by Figdraw.com). b , c Track plot of the elevated plus maze, and statistical analysis including percentage of time spent in the open arms, percentage of distance traveled in the open arms and percentage of times entering the open arms. n = 6 mice/group. d , e Track plot of the elevated zero maze, and statistical analysis including percentage of time spent in the open zones and percentage of times entering the open zones. n = 6 mice/group. f Statistical analysis, including percentage of distance traveled in the light area, percentage of time spent in the light area, and times of transitions in the light-dark box test. n = 6 mice/group. g Representative images of H&E-staining for the hippocampus sections. Scale bar = 100 μm. n = 6 independent experiments. h Representative images of Nissl staining for the hippocampus sections. Scale bar = 100 μm. n = 4 independent experiments. i The content of inositol in hippocampus tissue tested by ELISA. n = 8 mice/group. j The content of inositol in serum tested by ELISA. n = 8 mice/group. k–n The mRNA expression of TNF-α , IL-1β , IL-6 and iNOS in BV-2 cells. n = 3 mice/group. o, p The mRNA expression of Gabra3 and Camk2d in C8D1A cells. n = 3 mice/group. q The protein level of GABA A R, CaMKII, GABA B R detected by western blot. n = 3 independent experiments. Data were presented as means ± SD. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001. Source data are provided as a file.

Article Snippet: Next, primary antibody against GFAP (1:1000, A8335, 1:200, NB100-53809, USA), GABA A Rα1 (1:1000, 12708-1-AP, Proteintech, China), GABA B R2 (1:1000, 27567-1-AP, Proteintech, China), CaMKII (1:1000, 12716, Cell Signaling Technology, USA), GAD65 (1:1000, ab239372, Abcam, USA), CD206 (1:1000, ab64693, Abcam, USA), iNOS (1:1000, 18985-1-AP, Proteintech, China), CD80 (1:1000, 66406-1, Proteintech, China), p-p65 (1:1000, A00284-1, Boster, China), p-IKBα (1:1000, WLH3930, Wanleibio, China), and β-actin (50201, 1:1000, Kemei Borui Science and Technology, China) were incubated with the membranes overnight at 4 °C.

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot

Journal: International journal of molecular sciences

Article Title: Altered GABA A Receptor Expression in the Primary Somatosensory Cortex of a Mouse Model of Genetic Absence Epilepsy.

doi: 10.3390/ijms232415685

Figure Lengend Snippet: Figure 1. Representative confocal immunofluorescence images demonstrating expression of GABAAR α1 in the stargazer primary SoCx. 10× (Objective: 10× Plan) magnification from control (NE) littermate and epileptic (E) stargazer are shown here. (A) Pseudo-green channel shows diffuse labelling for GABAAR α1 subunit across the cortical layers with higher intensity in layer IV. (B) Pseudo-red channel shows labelling for PV+ neurons. The soma of PV+ neurons are present in all layers except layer I. (C) Pseudo-blue channel shows VGlut2 which assists in identification of the cortical layers. Intense labelling can be seen, predominantly, in layer IV due to the presence of thalamocortical excitatory nerve terminals.

Article Snippet: Membranes were probed using GABAAR α1 (1:500; AGA-001, Alomone, Jerusalem, Israel), GABAAR α3 (1:500; AGA-003, Alomone, Jerusalem, Israel), GABAAR α4 (1:200; AGA-008, Alomone, Jerusalem, Israel), GABAAR α5 (1:1000; AB9678, Sigma-Aldrich, St. Louis, MO, USA), GABAAR β2 (1:1000; ab8340, Abcam, Cambridge, UK), GABAAR β3 (1:1000; ab4046, Abcam, Cambridge, UK), GABAAR γ2 (1:200; AGA-005, Alomone, Jerusalem, Israel), and GABAAR δ (1:200; AGA014, Alomone, Jerusalem, Israel) with β-actin (1:1000; ab8226, Abcam, Cambridge UK) used Int.

Techniques: Confocal, Expressing, Control, Immunofluorescence

Journal: International journal of molecular sciences

Article Title: Altered GABA A Receptor Expression in the Primary Somatosensory Cortex of a Mouse Model of Genetic Absence Epilepsy.

doi: 10.3390/ijms232415685

Figure Lengend Snippet: Figure 2. Merged confocal immunofluorescence images demonstrating the co-labelling for GABAAR α1 and PV in the primary SoCx of control (NE) littermate and epileptic (E) stargazer. (A) 10× magnification of GABAAR α1/PV merged image shows co-localization of GABAAR α1 with PV+ somas and processes. (B) 60× (Objective: 60× PlanApo oil) magnification of GABAAR α1/PV merged image clearly shows GABAAR α1 present throughout the cortex as well on PV somas.

Article Snippet: Membranes were probed using GABAAR α1 (1:500; AGA-001, Alomone, Jerusalem, Israel), GABAAR α3 (1:500; AGA-003, Alomone, Jerusalem, Israel), GABAAR α4 (1:200; AGA-008, Alomone, Jerusalem, Israel), GABAAR α5 (1:1000; AB9678, Sigma-Aldrich, St. Louis, MO, USA), GABAAR β2 (1:1000; ab8340, Abcam, Cambridge, UK), GABAAR β3 (1:1000; ab4046, Abcam, Cambridge, UK), GABAAR γ2 (1:200; AGA-005, Alomone, Jerusalem, Israel), and GABAAR δ (1:200; AGA014, Alomone, Jerusalem, Israel) with β-actin (1:1000; ab8226, Abcam, Cambridge UK) used Int.

Techniques: Confocal, Control, Immunofluorescence

Journal: International journal of molecular sciences

Article Title: Altered GABA A Receptor Expression in the Primary Somatosensory Cortex of a Mouse Model of Genetic Absence Epilepsy.

doi: 10.3390/ijms232415685

Figure Lengend Snippet: Figure 3. WB analysis for GABAAR α1 and α3 subunits in the primary SoCx. Data compare expression between control (NE) littermates and epileptic (E) stargazers. (A) Representative blots display GABAAR α1 in the primary SoCx with β-actin as the loading control. Bar graphs represent the relative expression levels of GABAAR α1 (mean ± SEM). WB analysis revealed a statistically significant reduction in whole-tissue expression levels of GABAAR α1 in the primary SoCx of the stargazers (NE: 0.960 ± 0.066 [n = 18], E: 0.78 ± 0.046 [n = 14], p = 0.041); (B) Representative blots display GABAAR α3 in the primary SoCx with β-actin as the loading control. Bar graphs represent the relative expression levels of GABAAR α3. There was no statistically significant change in whole-tissue expression levels of GABAAR α3 in the primary SoCx of the stargazers (NE: 1.000 ± 0.036 [n = 17], E: 0.935 ± 0.049 [n = 15], p = 0.519). The significance threshold was set at 0.05 with * indicating p < 0.05. ‘ns’ indicates no significant change found from analyses.

Article Snippet: Membranes were probed using GABAAR α1 (1:500; AGA-001, Alomone, Jerusalem, Israel), GABAAR α3 (1:500; AGA-003, Alomone, Jerusalem, Israel), GABAAR α4 (1:200; AGA-008, Alomone, Jerusalem, Israel), GABAAR α5 (1:1000; AB9678, Sigma-Aldrich, St. Louis, MO, USA), GABAAR β2 (1:1000; ab8340, Abcam, Cambridge, UK), GABAAR β3 (1:1000; ab4046, Abcam, Cambridge, UK), GABAAR γ2 (1:200; AGA-005, Alomone, Jerusalem, Israel), and GABAAR δ (1:200; AGA014, Alomone, Jerusalem, Israel) with β-actin (1:1000; ab8226, Abcam, Cambridge UK) used Int.

Techniques: Analysis, Expressing, Control, Tissue, Western Blot

Journal: International journal of molecular sciences

Article Title: Altered GABA A Receptor Expression in the Primary Somatosensory Cortex of a Mouse Model of Genetic Absence Epilepsy.

doi: 10.3390/ijms232415685

Figure Lengend Snippet: Figure 4. WB analysis for GABAAR β2, β3, and γ2 subunits in the primary SoCx. The data compare expression between control (NE) littermates and epileptic (E) stargazers. (A–C) Representative blots display tissue expression of GABAAR β2, β3, and γ2 subunits in the primary SoCx with β-actin as the loading control. Bar graphs represent the relative expression levels of GABAAR β2, β3, and γ2 subunits, respectively. Bar graphs show a lack of statistically significant change in whole-tissue primary SoCx expression levels for β2 (NE: 0.917 ± 0.091 [n = 12], E: 0.678 ± 0.050 [n = 10]; p = 0.093), β3 (NE: 1.000 ± 0.067 [n = 12], E: 1.070 ± 0.109 [n = 10]; p = 0.859), and γ2 (NE: 1.000 ± 0.159 [n = 9], E: 0.946 ± 0.079 [n = 7]; p = 0.900). The significance threshold was set at 0.05. ‘ns’ indicates no significant change found from analyses.

Article Snippet: Membranes were probed using GABAAR α1 (1:500; AGA-001, Alomone, Jerusalem, Israel), GABAAR α3 (1:500; AGA-003, Alomone, Jerusalem, Israel), GABAAR α4 (1:200; AGA-008, Alomone, Jerusalem, Israel), GABAAR α5 (1:1000; AB9678, Sigma-Aldrich, St. Louis, MO, USA), GABAAR β2 (1:1000; ab8340, Abcam, Cambridge, UK), GABAAR β3 (1:1000; ab4046, Abcam, Cambridge, UK), GABAAR γ2 (1:200; AGA-005, Alomone, Jerusalem, Israel), and GABAAR δ (1:200; AGA014, Alomone, Jerusalem, Israel) with β-actin (1:1000; ab8226, Abcam, Cambridge UK) used Int.

Techniques: Analysis, Expressing, Control, Tissue, Western Blot

Journal: International journal of molecular sciences

Article Title: Altered GABA A Receptor Expression in the Primary Somatosensory Cortex of a Mouse Model of Genetic Absence Epilepsy.

doi: 10.3390/ijms232415685

Figure Lengend Snippet: Figure 5. WB analysis for the principal tonic GABAAR α4, α5, and δ subunits. Analysis compares the primary SoCx of control (NE) littermates and the epileptic (E) stargazers. (A–C) Representative blots display tissue expression of GABAAR α4, α5, and δ subunits in the primary SoCx with β-actin as the loading control. Bar graphs represent the relative expression levels of GABAAR α4, α5 and δ subunits, respectively. Bar graphs reveal a lack of statistically significant change in whole-tissue primary SoCx expression levels for GABAAR α4 (NE: 1.000 ± 0.063 [n = 18], E: 1.117 ± 0.095 [n = 15]; p = 0.325), α5 (NE: 1.000 ± 0.048 [n = 23], E: 0.937 ± 0.083 [n = 13]; p = 0.672), and δ (NE: 1.000 ± 0.041 [n = 27], E: 0.899 ± 0.069 [n = 23]; p = 0.215). The significance threshold was set at 0.05. ‘ns’ indicates no significant change found from analyses.

Article Snippet: Membranes were probed using GABAAR α1 (1:500; AGA-001, Alomone, Jerusalem, Israel), GABAAR α3 (1:500; AGA-003, Alomone, Jerusalem, Israel), GABAAR α4 (1:200; AGA-008, Alomone, Jerusalem, Israel), GABAAR α5 (1:1000; AB9678, Sigma-Aldrich, St. Louis, MO, USA), GABAAR β2 (1:1000; ab8340, Abcam, Cambridge, UK), GABAAR β3 (1:1000; ab4046, Abcam, Cambridge, UK), GABAAR γ2 (1:200; AGA-005, Alomone, Jerusalem, Israel), and GABAAR δ (1:200; AGA014, Alomone, Jerusalem, Israel) with β-actin (1:1000; ab8226, Abcam, Cambridge UK) used Int.

Techniques: Analysis, Control, Tissue, Expressing, Western Blot

Journal: International journal of molecular sciences

Article Title: Altered GABA A Receptor Expression in the Primary Somatosensory Cortex of a Mouse Model of Genetic Absence Epilepsy.

doi: 10.3390/ijms232415685

Figure Lengend Snippet: Figure 6. Pilot WB run for biochemically isolated primary SoCx subcellular fractions (total lysate, cytosol, extra-synaptic and synaptic). This run confirmed the successful isolation of the synaptic fractions given the intense labelling for both PSD95 and GABAAR α1. Extra-synaptic fraction, on the other hand, showed no bands for PSD95, but low intensity bands were seen for GABAAR α1 and β-actin. PanC showed good expression in all subcellular fractions.

Article Snippet: Membranes were probed using GABAAR α1 (1:500; AGA-001, Alomone, Jerusalem, Israel), GABAAR α3 (1:500; AGA-003, Alomone, Jerusalem, Israel), GABAAR α4 (1:200; AGA-008, Alomone, Jerusalem, Israel), GABAAR α5 (1:1000; AB9678, Sigma-Aldrich, St. Louis, MO, USA), GABAAR β2 (1:1000; ab8340, Abcam, Cambridge, UK), GABAAR β3 (1:1000; ab4046, Abcam, Cambridge, UK), GABAAR γ2 (1:200; AGA-005, Alomone, Jerusalem, Israel), and GABAAR δ (1:200; AGA014, Alomone, Jerusalem, Israel) with β-actin (1:1000; ab8226, Abcam, Cambridge UK) used Int.

Techniques: Isolation, Expressing, Western Blot, Cell Fractionation

Journal: International journal of molecular sciences

Article Title: Altered GABA A Receptor Expression in the Primary Somatosensory Cortex of a Mouse Model of Genetic Absence Epilepsy.

doi: 10.3390/ijms232415685

Figure Lengend Snippet: Figure 7. WB analyses of biochemically isolated fractions from the primary SoCx for GABAAR α1 and α3 subunits. Isolated subcellular fractions were compared between control (NE) litter- mates and epileptic (E) stargazers. (A–D) Biochemical fractionation analysis revealed a 12.2% reduction in the synaptic expression of phasic GABAAR α1 in the primary SoCx of the stargaz- ers compared to their control littermates (NE: 1.000 ± 0.146 [n = 10], E: 0.878 ± 0.037 [n = 10]; p = 0.002); no significant difference was revealed from the other subcellular components: total lysate (NE: 1.000 ± 0.043 [n = 7], E: 0.961 ± 0.052 [n = 6]; p = 0.531), cytosol (NE: 1.000 ± 0.034 [n = 7], E: 1.100 ± 0.098 [n = 6]; p = 0.443), and extra-synaptic (NE: 1.000 ± 0.074 [n = 9], E: 0.912 ± 0.094 [n = 9]; p = 0.385). (E–H) Biochemical fractionation analysis revealed no significant change in GABAAR α3 in all subcellular components from the primary SoCx of stargazers compared to their control littermates: Total lysate (NE: 1.000 ± 0.026 [n = 12], E: 1.035 ± 0.060 [n = 10]; p = 0.381), cytosol (NE: 1.000 ± 0.037 [n = 11], E: 1.181 ± 0.118 [n = 10]; p = 0.349), extra-synaptic (NE: 1.000 ± 0.031 [n = 11], E: 1.093 ± 0.125 [n = 11]; p = 0.133), and synaptic (NE: 0.967 ± 0.034 [n = 11], E: 1.205 ± 0.153 [n = 9], p = 0.359). The sig- nificance threshold was set at 0.05 with ** indicating p < 0.01. ‘ns’ indicates no significant change found from analyses.

Article Snippet: Membranes were probed using GABAAR α1 (1:500; AGA-001, Alomone, Jerusalem, Israel), GABAAR α3 (1:500; AGA-003, Alomone, Jerusalem, Israel), GABAAR α4 (1:200; AGA-008, Alomone, Jerusalem, Israel), GABAAR α5 (1:1000; AB9678, Sigma-Aldrich, St. Louis, MO, USA), GABAAR β2 (1:1000; ab8340, Abcam, Cambridge, UK), GABAAR β3 (1:1000; ab4046, Abcam, Cambridge, UK), GABAAR γ2 (1:200; AGA-005, Alomone, Jerusalem, Israel), and GABAAR δ (1:200; AGA014, Alomone, Jerusalem, Israel) with β-actin (1:1000; ab8226, Abcam, Cambridge UK) used Int.

Techniques: Isolation, Control, Fractionation, Analysis, Expressing, Western Blot

Journal: International journal of molecular sciences

Article Title: Altered GABA A Receptor Expression in the Primary Somatosensory Cortex of a Mouse Model of Genetic Absence Epilepsy.

doi: 10.3390/ijms232415685

Figure Lengend Snippet: Figure 8. WB analyses of biochemically isolated fractions from primary SoCx for GABAAR α4, α5, and δ subunits. Subcellular fractions were isolated from the control (NE) littermates and epileptic epileptic (E) stargazers. (A–D) Biochemical fractionation analysis revealed no signif- icant change in GABAAR α4 in all subcellular components from the primary SoCx of stargaz- ers compared to their NE littermates: total lysate (NE: 1.000 ± 0.063 [n = 9], E: 1.353 ± 0.227 [n = 9]; p = 0.257), cytosol (NE: 1.000 ± 0.088 [n = 14], E: 1.308 ± 0.223 [n = 10]; p = 0.172), extra-synaptic (NE: 1.000 ± 0.051 [n = 14], E: 0.789 ± 0.183 [n = 9]; p = 0.516), and synaptic (NE: 1.000 ± 0.047 [n = 14], E: 0.955 ± 0.122 [n = 9]; p = 0.369). (E–H) GABAAR α5 anal- ysis revealed no significant change in all subcellular components from the primary SoCx of stargazers compared to their control littermates: total lysate (NE: 1.000 ± 0.094 [n = 10], E: 0.919 ± 0.102 [n = 10]; p = 0.566), cytosol (NE: 1.000 ± 0.113 [n = 14], E: 1.295 ± 0.362 [n = 8]; p = 0.920), extra-synaptic (NE: 1.000 ± 0.068 [n = 13], E: 1.324 ± 0.214 [n = 9]; p = 0.431) and, synaptic (NE: 1.000 ± 0.075 [n = 13], E: 1.190 ± 0.195 [n = 9]; p = 0.744). (I–L) No statistically significant change was also seen for GABAAR δ subunit in all subcellular compo- nents from the primary SoCx of stargazers compared to their control littermates: total lysate (NE:1.000 ± 0.071 [n = 15], E: 1.088 ± 0.201 [n = 9]; p = 0.815), cytosol (NE: 1.000 ± 0.083 [n = 15], E: 0.831 ± 0.138 [n = 9]; p = 0.379), extra-synaptic (NE: 1.000 ± 0.078 [n = 15], E: 1.419 ± 0.241 [n = 10]; p = 0.191), and synaptic (NE: 1.000 ± 0.053 [n = 15], E: 1.092 ± 0.094 [n = 9]; p = 0.263). The significance threshold was set at 0.05. ‘ns’ indicates no significant change found from analyses.

Article Snippet: Membranes were probed using GABAAR α1 (1:500; AGA-001, Alomone, Jerusalem, Israel), GABAAR α3 (1:500; AGA-003, Alomone, Jerusalem, Israel), GABAAR α4 (1:200; AGA-008, Alomone, Jerusalem, Israel), GABAAR α5 (1:1000; AB9678, Sigma-Aldrich, St. Louis, MO, USA), GABAAR β2 (1:1000; ab8340, Abcam, Cambridge, UK), GABAAR β3 (1:1000; ab4046, Abcam, Cambridge, UK), GABAAR γ2 (1:200; AGA-005, Alomone, Jerusalem, Israel), and GABAAR δ (1:200; AGA014, Alomone, Jerusalem, Israel) with β-actin (1:1000; ab8226, Abcam, Cambridge UK) used Int.

Techniques: Isolation, Control, Fractionation, Analysis, Western Blot

Journal: International journal of molecular sciences

Article Title: Altered GABA A Receptor Expression in the Primary Somatosensory Cortex of a Mouse Model of Genetic Absence Epilepsy.

doi: 10.3390/ijms232415685

Figure Lengend Snippet: Figure 9. ICC-EM analysis for GABAAR α1 at PV+ and non-PV inhibitory synapses in the primary SoCx compared and analyzed between control (NE) littermates and epileptic (E) stargazers. From top to bottom: Representative EM micrographs demonstrate PV labelling in the presynaptic terminals (pre), which appear darker stained, in stargazer and NE littermate primary SoCx. Arrow represents 20 nm immunogold particle labelling PV while arrowhead represents 10 nm immunogold labelling GABAAR α1. (A) For analysis for GABAAR α1 at inhibitory synapses from non-PV terminals onto non-PV profiles, as indicated by lack of 20 nm gold particles, 490 synapses in stargazers and 490 in control littermates primary SoCx were analyzed from seven pairs distributed equally. No significant difference was seen in the seven pairs (NE: 10.65 ± 0.918 [n = 7], E: 10.72 ± 0.751 [n = 7]; p = 0.804). (B) Analysis for GABAAR α1 at inhibitory synapses with PV (20 nm gold) in the presynaptic terminals revealed no significant difference between the stargazers (63 synapses analyzed) and control littermates (63 synapses analyzed) (NE: 11.82 ± 1.325 [n = 7], E: 11.15 ± 1.007 [n = 7], p > 0.999). The significance threshold was set at 0.05. ‘ns’ indicates no significant change found from analyses.

Article Snippet: Membranes were probed using GABAAR α1 (1:500; AGA-001, Alomone, Jerusalem, Israel), GABAAR α3 (1:500; AGA-003, Alomone, Jerusalem, Israel), GABAAR α4 (1:200; AGA-008, Alomone, Jerusalem, Israel), GABAAR α5 (1:1000; AB9678, Sigma-Aldrich, St. Louis, MO, USA), GABAAR β2 (1:1000; ab8340, Abcam, Cambridge, UK), GABAAR β3 (1:1000; ab4046, Abcam, Cambridge, UK), GABAAR γ2 (1:200; AGA-005, Alomone, Jerusalem, Israel), and GABAAR δ (1:200; AGA014, Alomone, Jerusalem, Israel) with β-actin (1:1000; ab8226, Abcam, Cambridge UK) used Int.

Techniques: Analysis, Control, Staining, Electron Microscopy