Journal: International Journal of Molecular Sciences

Article Title: GABAB-Dependent Negative Feedback Contributes to Earlier Epileptiform Discharge Termination Within Malformed Cortex

doi: 10.3390/ijms27177896

Figure Lengend Snippet: GABA B receptor modulation affects epileptiform discharge duration in control and malformed cortex. ( A ) Local application of the GABA B receptor agonist SKF-97541 (5 µM, 3 s) induces a slow hyperpolarization in current-clamp (left, pipette solution 1) and an outward current in voltage-clamp (right, pipette solution 2; n = 5 cells from N = 2 animals). ( B ) Bath application of CGP-55845 (3 µM) transforms brief discharges into prolonged multi-peak events. ( C ) Expanded view from ( B ) (gray boxes). CGP-55845 prolongs discharge duration and alters the I PD . ( D ) Expanded view from ( B ) (orange lines) showing comparable initiation but divergent termination. Panels ( B – D ) are representative of n = 15 recordings from N = 5 control animals. ( E ) CGP-55845 transforms spontaneous activity in the FFL slice (recording configuration as in B). Blue lines mark regions expanded in ( F ). ( F ) Expanded view illustrating transformation to prolonged ictal-like events; n = 12 recordings from N = 4 FFL animals.

Article Snippet: Pharmacological agents were applied as follows: the GABA B receptor antagonist CGP-55845 (3 μM; Alomone Labs, Jerusalem, Israel) was bath-applied from a DMSO stock solution (final DMSO concentration ≤ 0.1%), with a minimum wash-in period of 6 min.

Techniques: Control, Transferring, Activity Assay, Transformation Assay

Journal: International Journal of Molecular Sciences

Article Title: GABAB-Dependent Negative Feedback Contributes to Earlier Epileptiform Discharge Termination Within Malformed Cortex

doi: 10.3390/ijms27177896

Figure Lengend Snippet: The effect of GABA B receptor blockade on evoked epileptiform discharges. ( A ) Voltage-clamp recordings in control slices before and after CGP-55845 application (Vh = −20 mV). ( B ) Discharge duration before and after CGP-55845 application in control slices ( n = 11 paired recordings, N = 5; paired t -test). ( C ) Schematic showing the stimulating electrode within the microgyrus and recording pipettes positioned in the microgyrus and PMZ. ( D ) Superimposed recordings from both locations before and after CGP-55845 (normalized to peak). ( E ) Statistical comparison of evoked discharge duration (two-way repeated-measures ANOVA, n = 7 pairs, N = 4). ( F ) Left: averaged recordings (normalized, Vh = −20 mV); gray area represents SD. Right: normalized current decay slopes (Slopenorm; two-way repeated-measures ANOVA, n = 7 pairs, N = 4). * p < 0.05, ** p < 0.01; ns, not significant; Sidak’s post hoc test.

Article Snippet: Pharmacological agents were applied as follows: the GABA B receptor antagonist CGP-55845 (3 μM; Alomone Labs, Jerusalem, Israel) was bath-applied from a DMSO stock solution (final DMSO concentration ≤ 0.1%), with a minimum wash-in period of 6 min.

Techniques: Control, Comparison

Journal: Signal Transduction and Targeted Therapy

Article Title: Endothelial GABA protects against aortic dissection by inhibiting endothelial and mitochondrial dysfunction and maintaining vascular homeostasis

doi: 10.1038/s41392-026-02619-2

Figure Lengend Snippet: GABA maintains homeostatic mitochondrial levels in the context of oxidative stress. a , b Representative images and quantification of western blots of ICAM-1, P-p42, total-p42, and c-FOS expression in HAECs treated with GABA or GABA + CGP52432 (one-way ANOVA, mean ± S.D.; n = 4). c Representative morphology of the mitochondrial network structure as assayed by MitoTracker Red staining in HAECs treated with GABA alone or in combination with CGP52432 . d , e Mitochondrial ROS production in HAECs treated with GABA alone or in combination with CGP52432 (one-way ANOVA, mean ± S.D.; n = 3). Scale bar = 10 μm. f Intercellular ATP synthesis in H 2 O 2 -induced HAECs treated with GABA alone or in combination with CGP52432 (one-way ANOVA, mean ± S.D.; n = 3). g The impact of GABA on oxidative phosphorylation and glycolytic rates in H 2 O 2 -induced HAECs. h Western blotting of GABBR2 accumulation in mitochondria with GABA regulation according to different time spans. i Western blotting of GABBR2 accumulation in mitochondria in the GABA- or CGP52432 -treated groups. j , k Representative images and quantification of the correlations of GABBR2 and mitochondria with GABA alone or in combination with CGP52432 (one-way ANOVA, mean ± S.D.). Scale bar = 5 μm

Article Snippet: The GABA B receptor antagonist CGP52432 (MCE, USA; catalog: HY-103531; intravenous injection once daily, 10 mg/kg/dose) was administered.

Techniques: Western Blot, Expressing, Staining, Phospho-proteomics

Journal: Signal Transduction and Targeted Therapy

Article Title: Endothelial GABA protects against aortic dissection by inhibiting endothelial and mitochondrial dysfunction and maintaining vascular homeostasis

doi: 10.1038/s41392-026-02619-2

Figure Lengend Snippet: Endothelial cell-derived GABA protects smooth muscle cell homeostasis from inflammatory stimulation. a Representative images and quantification of western blot showing the protein expression of MMP-2 and MMP-9 in IL-1b-induced SMCs treated with GABA alone or in combination with CGP52432 (one-way ANOVA, mean ± S.D.; n = 3). b qRT‒PCR analysis of the mRNA expression of MMP-2 and MMP-9 in IL-1b-induced SMCs treated with GABA alone or in combination with CGP52432 (one-way ANOVA, mean ± S.D.; n = 3). c The differentially expressed proteins in IL-1b-induced SMCs with and without GABA treatment are shown as volcano plots. d Heatmap analysis indicates that Notch3 participates in this process. e Schematic diagrams of the EC/SMC coculture system. ECs and SMCs were seeded on the upper and lower sides of a membrane, respectively. f , g Western blot showing the expression of Notch3, MMP-2, and CNN1 in IL-1b-induced SMCs cocultured with ECs (one-way ANOVA, mean ± S.D.; n = 3). h , i Representative immunofluorescence images of Notch3 expression in the aortic wall of BAPN-induced AAV EC-OE GAD1 mice and vehicle-treated mice ( t test, mean ± S.D.; n = 8, 2 visual fields per sample). Scale bar = 50 μm

Article Snippet: The GABA B receptor antagonist CGP52432 (MCE, USA; catalog: HY-103531; intravenous injection once daily, 10 mg/kg/dose) was administered.

Techniques: Derivative Assay, Western Blot, Expressing, Membrane, Immunofluorescence

Journal: Signal Transduction and Targeted Therapy

Article Title: Endothelial GABA protects against aortic dissection by inhibiting endothelial and mitochondrial dysfunction and maintaining vascular homeostasis

doi: 10.1038/s41392-026-02619-2

Figure Lengend Snippet: Rewiring GABA metabolism reverses vascular homeostasis disorders. a Representative morphology of aortas from the BAPN, GABA 2-week, and GABA + CGP52432 2-week groups. Scale bar = 2 mm. b Incidence of TAD in the BAPN, GABA 2-week, and GABA + CGP52432 2-week groups ( n = 16). c Probability of survival in the BAPN, GABA 2-week, and GABA + CGP52432 2-week groups (log-rank (Mantel‒Cox) test; n = 16). d , e Permeation of Evans blue dye into the thoracic aortas of the BAPN, GABA 2-week, and GABA + CGP52432 2-week groups (one-way ANOVA, mean ± S.D.; n = 3). f , g Representative EVG staining and quantification of elastin degradation in the BAPN, GABA 2-week, and GABA + CGP52432 2-week groups (one-way ANOVA, mean ± S.D.; n = 16). Scale bar = 20 μm. ( h – i ) Representative en face staining of ICAM-1 and statistical data for the aortic wall (one-way ANOVA, mean ± S.D.; n = 6; 2 visual fields per sample). Scale bar = 10 μm. j , k Representative en face staining of c-FOS and statistical data for the aortic wall from differently treated mice ( n = 6, 2 visual fields per sample). Scale bar = 10 μm. l Representative immunofluorescence staining of infiltrating macrophages (F4/80) in the aortic wall of mice subjected to different treatments. Scale bar = 50 μm. m Representative immunofluorescence images of Notch3 expression in the aortic wall of mice subjected to different treatments. Scale bar = 50 μm

Article Snippet: The GABA B receptor antagonist CGP52432 (MCE, USA; catalog: HY-103531; intravenous injection once daily, 10 mg/kg/dose) was administered.

Techniques: Staining, Immunofluorescence, Expressing

Journal: Nature Communications

Article Title: Circulating extracellular vesicle microRNAs mediate immune modulation of social behavior in male mice

doi: 10.1038/s41467-026-70469-1

Figure Lengend Snippet: a Flow of data analysis to identify candidate genes responsible for GABAergic signaling changes in Rag1 −/− mPFC. b Representative confocal microscope images showing enhanced neuronal PKCε immunoreactivity in the mPFC of Rag1 −/− mice. c Quantification of PKCε + neurons in the mPFC of WT and Rag1 −/− mice. WT mice, n = 7 Rag1 −/− mice, n = 6. d Representative confocal microscope images of Gephyrin and GABA A Rγ2 co-localization in CaMKIIα + neurons of WT and Rag1 −/− mPFC. e Quantification of number of overlapped puncta of Gephyrin and GABA A Rγ2 in CaMKIIα + neurons of WT and Rag1 −/− mPFC. WT mice, n = 7; Rag1 −/− mice, n = 6. f Expression levels of miR-23a-3p and miR-103-3p in the bEVs from WT and Rag1 −/− mice ( n = 8 per group). Data are shown as fold-change relative to WT data. Scale bar, 50 μm. Each bar graph represents mean ± SEM. Each dot represents one mouse. ns, not significant. * p < 0.05, *** p < 0.005. Significance was determined by Student’s t -test. See Supplementary Data for details of the statistical analysis. Source data are provided as a Source Data file.

Article Snippet: The sections were then placed in blocking solution (PBS supplemented with 2% Normal Goat Serum, 1% BSA, 0.1% TritonX, 0.05% Tween-20, and 0.05% sodium azide) for 1 h at room temperature and then incubated at 4 °C overnight with the following primary antibodies: mouse anti-NeuN (1:500, #MAB377, Merck-Millipore, RRID: AB_2298772), rabbit anti-Iba1 (1:400, #019-19741, Wako Chemicals, RRID: AB_839504), goat anti-c-Fos (1:500, #sc-52, Santa Cruz Biotechnology, RRID: AB_2106783), rabbit anti-c-Fos (1:200, #2250, Cell Signaling, RRID: AB_22472111), mouse anti-CaMKIIα (1:200, #688602, Clone 6G9, BioLegend, RRID: AB_2617027), rabbit anti-CaMKIIα (1:100, #20666-1-AP, Proteintech, RRID:AB_2878722), rabbit anti-RFP (1:1000, #600-401-379, Rockland, RRID: AB_2209751), rabbit anti-PKCε (1:100, #20877-1-AP, Proteintech, RRID:AB_10697812), mouse anti-CC1 (1:100, #OP80, Calbiochem, RRID:AB_2057371), rabbit anti-S100β (1:200, #ab868, Abcam, RRID:AB_306716), mouse anti-NSE (1:500, #66150-1-g, Proteintech, RRID:AB_2881546), mouse anti-Gephyrin (1:100, #147021, Synaptic Systems, RRID:AB_2232546), and rabbit anti-GABA A Rγ2 (1:100, #AGA-005, Alomone Labs, RRID: AB_2039870).

Techniques: Analysis, Confocal, Microscopy, Expressing, Immunofluorescence

Journal: Nature Communications

Article Title: Circulating extracellular vesicle microRNAs mediate immune modulation of social behavior in male mice

doi: 10.1038/s41467-026-70469-1

Figure Lengend Snippet: a Experimental outline of bEV collection, miRNA mimics loading, intravenous injection, and subsequent behavioral assay. b Sociability data of Rag1 −/− mice administered with Rag1 - /- bEVs containing negative control miRNA mimics (control mimics) or miRNA mimics for miR-23a-3p and miR-103-3p (miR-23a-3p + miR-103-3p mimics). Control, n = 15; miR-23a-3p + miR-103-3p, n = 11. c Representative confocal microscope images of neuronal c-Fos expression in the mPFC of Rag1 −/− mice administered with Rag1 -/- bEVs containing control or miR-23a-3p + miR-103-3p mimics ( n = 10 mice per group). d Quantification of c-Fos + neurons in the mPFC ( n = 10 per group). e Representative confocal microscope images of neuronal PKCε expression in the mPFC of Rag1 −/− mice administered with Rag1 -/- bEVs containing control or miR-23a-3p + miR-103-3p mimics. f Quantification of PKCε + neurons in the mPFC ( n = 10 per group). g Representative confocal microscope images of neuronal Gephyrin and GABA A Rγ2 (expression in CaMKIIα + neurons in the mPFC of Rag1 −/− mice administered with Rag1 -/- bEVs containing control or miR-23a-3p + miR-103-3p mimics. h Quantification of number of Gephyrin and GABA A Rγ2 puncta in CaMKIIα + neurons in the mPFC. Control, n = 10; miR-23a-3p + miR-103-3p, n = 11. * p < 0.0001. i Representative traces of sIPSCs of the mPFC neurons in brain slices from Rag1 −/− mice administered with Rag1 -/- bEVs containing control or miR-23a-3p + miR-103-3p mimics. j Quantification data of sIPSC amplitude and frequency changes by miR-23a-3p + miR-103-3p mimics in Rag1 −/− mice (aggregated data from n = 3 mice per group). k Graphical summary of mechanism. Circulating EV miRNAs, including miR-23a-3p and miR-103-3p, inhibit the expression of PKCε in mPFC neurons, increasing synaptic GABA A receptor localization and enhancing inhibitory postsynaptic signaling. Scale bars, 50 μm ( c , e ) and 10 μm ( i ). Each bar graph represents mean ± SEM. Each dot represents one mouse ( b , d , f , h ) or cell ( j ). ns, not significant. *p < 0.05, **p < 0.01, and ***p < 0.005. Significance was determined by Student’s t -test. See Supplementary Data for details of the statistical analysis. Source data are provided as a Source Data file.

Article Snippet: The sections were then placed in blocking solution (PBS supplemented with 2% Normal Goat Serum, 1% BSA, 0.1% TritonX, 0.05% Tween-20, and 0.05% sodium azide) for 1 h at room temperature and then incubated at 4 °C overnight with the following primary antibodies: mouse anti-NeuN (1:500, #MAB377, Merck-Millipore, RRID: AB_2298772), rabbit anti-Iba1 (1:400, #019-19741, Wako Chemicals, RRID: AB_839504), goat anti-c-Fos (1:500, #sc-52, Santa Cruz Biotechnology, RRID: AB_2106783), rabbit anti-c-Fos (1:200, #2250, Cell Signaling, RRID: AB_22472111), mouse anti-CaMKIIα (1:200, #688602, Clone 6G9, BioLegend, RRID: AB_2617027), rabbit anti-CaMKIIα (1:100, #20666-1-AP, Proteintech, RRID:AB_2878722), rabbit anti-RFP (1:1000, #600-401-379, Rockland, RRID: AB_2209751), rabbit anti-PKCε (1:100, #20877-1-AP, Proteintech, RRID:AB_10697812), mouse anti-CC1 (1:100, #OP80, Calbiochem, RRID:AB_2057371), rabbit anti-S100β (1:200, #ab868, Abcam, RRID:AB_306716), mouse anti-NSE (1:500, #66150-1-g, Proteintech, RRID:AB_2881546), mouse anti-Gephyrin (1:100, #147021, Synaptic Systems, RRID:AB_2232546), and rabbit anti-GABA A Rγ2 (1:100, #AGA-005, Alomone Labs, RRID: AB_2039870).

Techniques: Injection, Behavioral Assay, Negative Control, Control, Confocal, Microscopy, Expressing, Analysis, Immunofluorescence, Patch Clamp