Journal: eLife

Article Title: Perisomatic GABAergic synapses of basket cells effectively control principal neuron activity in amygdala networks

doi: 10.7554/eLife.20721

Figure Lengend Snippet: ( A ) Schematic drawing of the experiment determining the area affected by local somatic gabazine application. ( B ) Representative recordings of the GABA puff evoked currents at various dendritic locations in control conditions (black) and upon somatic gabazine application (red). ( C ) Summary of recordings from 22 dendritic locations show that the somatic gabazine puff can significantly reduce GABAergic currents (red dots) in an area of 50 µm radius from the site of the application, which covers the perisomatic region. ( D ) Schematic drawing of the experiment measuring the effect of the elimination of perisomatic contacts by local somatic gabazine application. ( E and F ) Representative experiment showing the rapid and substantial drop in IPSC charge transfer upon gabazine application (applied for 75 s in panel F). ( G and H ) Elimination of perisomatic inhibitory inputs resulted in a significant suppression of the IPSC charge transfer and inhibitory efficacy (p=0.01 and p=0.007, respectively, Wilcoxon Signed Rank test). Blue: CCKBC-PN pairs, orange: PVBC-PN pairs. Open circles represent recorded pairs, filled circles represent median values. ( I and J ) Relationship between the inhibitory efficacy and IPSC charge transfer ( I ) or the number of functioning perisomatic contacts ( J ) before (black dots) and after (red dots) the pharmacological elimination of perisomatic inhibition (individual experiments connected with red line). Grey dots show data from previous recordings for comparison (data from and ). Scales: B: 20 pA and 50 ms; 20 µm, E: 8 pA and 20 ms. DOI: http://dx.doi.org/10.7554/eLife.20721.015

Article Snippet: To block GABAergic synaptic transmission locally at the perisomatic region of PNs we applied a GABA A antagonist gabazine (1 μM, Tocris Bioscience) with a standard patch pipette on the cell body of the PN by continuous pressure, during whole cell recording of a monosynaptically connected interneuron-PN pair as described above.

Techniques: Control, Inhibition, Comparison

Journal: The Journal of Physiology

Article Title: GABA_B‐ and GABA_A‐receptor‐mediated regulation of Up and Down states across development

doi: 10.1113/jp282736

Figure Lengend Snippet: Figure 5. Quantification of MUA signal and evaluation of LFP event classification as either Up states or SWDs A, LFP events were detected and their onsets/offsets were marked using our previously published software (LFPAnalyzer; Tsakanikas et al., 2017; top panels); the calculated duration of each event (either Up state (a) or SWD (b)) is indicated by the grey boxes. Subsequently, the MUA of all events was detected (middle panel) and quantified (lower panel), as described in Methods. B, scatter plots of Up states (either in control condition, filled circles, or under gabazine application, filled diamonds) and SWDs (open squares), using peak amplitude and MUA

Article Snippet: After recording spontaneous cortical activity in normal aCSF for 20 min, the perfusate was replaced with aCSF containing the GABAA antagonist gabazine (200 nm; SR95531 hydrobromide; Tocris Bioscience, Bristol, UK) and monitored for another 40–60 min.

Techniques: Software, Control

Journal: The Journal of Physiology

Article Title: GABA_B‐ and GABA_A‐receptor‐mediated regulation of Up and Down states across development

doi: 10.1113/jp282736

Figure Lengend Snippet: Figure 6. Raw traces of simultaneous intra- (IC) and extracellular (LFP) recordings from young (red traces) and adult (blue traces) mice in control and gabazine conditions A, high temporal resolution traces of individual events. Red (young) and blue (adult) continuous lines above the events indicate the Up state duration as measured in the LFP signal. Similarly, red (young) and blue (adult) dotted lines indicate the duration of SWDs recorded under gabazine condition. B, low temporal resolution recording traces, indicative of the Up state rate of occurrence. Continuous-lined and dashed-lined rectangles indicate the presence of either Up states or SWDs, respectively. Gabazine led to the emergence of SWDs in both ages but decreased the occurrence of Up states only in young animals. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: After recording spontaneous cortical activity in normal aCSF for 20 min, the perfusate was replaced with aCSF containing the GABAA antagonist gabazine (200 nm; SR95531 hydrobromide; Tocris Bioscience, Bristol, UK) and monitored for another 40–60 min.

Techniques: Control

Journal: The Journal of Physiology

Article Title: GABA_B‐ and GABA_A‐receptor‐mediated regulation of Up and Down states across development

doi: 10.1113/jp282736

Figure Lengend Snippet: Figure 7. Impact of GABAA-mediated inhibition on spontaneous cortical activity in young (n (N) = 11 (8)) and adult (n (N) = 11 (11)) networks A, split violin plots showing the effect of GABAA blocker (gabazine 200 nM) on Up state duration: reduction in young (control: 1.88 (0.38) s vs. gabazine: 0.49 (0.59) s, P = 0.0003), but no effect in adults (control: 1.21 (0.38) s vs. gabazine: 1.12 (0.30) s, P = 0.3480). Black lines indicate the overall mean Up state duration and each circle indicates the average values from a brain slice. B, box-plots for Up state rate of occurrence. Gabazine decreased dramatically the metric in young (control: 0.039 (0.020) Hz vs. gabazine: 0.002 (0.002) Hz, P = 0.0001) but did not

Article Snippet: After recording spontaneous cortical activity in normal aCSF for 20 min, the perfusate was replaced with aCSF containing the GABAA antagonist gabazine (200 nm; SR95531 hydrobromide; Tocris Bioscience, Bristol, UK) and monitored for another 40–60 min.

Techniques: Inhibition, Activity Assay, Control, Slice Preparation

Journal: The Journal of Physiology

Article Title: GABA_B‐ and GABA_A‐receptor‐mediated regulation of Up and Down states across development

doi: 10.1113/jp282736

Figure Lengend Snippet: Figure 8. Cluster analysis of Up state activity in young and adult animals A Gaussian mixture model clustering analysis was conducted in two dimensions (Up state duration and Up state interval) for the three experimental conditions: control (A), CGP (B) and gabazine (C). In all cases the best fit number of GMM components, calculated according to Calinski–Harabasz and silhouette criteria, revealed two clusters. Red and blue data points represent individual Up state events obtained from young (control: 2016 events from n (N) = 22 (15), CGP: 330 events from n (N) = 11 (7), gabazine: 19 events from n (N) = 11 (8)) and adult (control: 1030 events from n (N) = 22 (22), CGP: 318 events from n (N) = 11 (11), gabazine: 351 events from n (N) = 11 (11)) mice, respectively. The ellipses correspond to multiples of standard deviation (σ) for the 2D Gaussian distribution. Starting from the inner one, the four ellipses represent 0.5σ, 1σ, 1.5σ and 2σ. 1σ ellipse covers ∼39%, whereas 2σ ellipse covers ∼86.5% of the total probability mass for the standard bivariate Gaussian distribution. Mean values of these datasets are also shown in rotating graphs. For a rotating 3D version of B, see Supporting information, Video S2; for a rotating 3D version of C, see Supporting information, Video S3. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: After recording spontaneous cortical activity in normal aCSF for 20 min, the perfusate was replaced with aCSF containing the GABAA antagonist gabazine (200 nm; SR95531 hydrobromide; Tocris Bioscience, Bristol, UK) and monitored for another 40–60 min.

Techniques: Activity Assay, Control, Standard Deviation

Journal: The Journal of Physiology

Article Title: GABA_B‐ and GABA_A‐receptor‐mediated regulation of Up and Down states across development

doi: 10.1113/jp282736

Figure Lengend Snippet: Figure 9. Effect of non-saturating concentrations of gabazine on phasic GABA currents in young and adult animals A–C, features of mIPSCs (peak amplitude, A; decay time 90–10, B; decay τ w, C) are affected by 200 nM gabazine in an age-independent way. As statistical analysis revealed no significant interaction between age and drug, data from young (n (N) = 5 (5)) and adult (n (N) = 4 (4)) animals are pooled to report main effect of gabazine. Individual data points in red and blue indicate mean values for young and adult animals, respectively. (for A, control: −57.09 (17.32) pA vs. gabazine: −16.61 (13.43) pA, P = 0.0013; for B, control: 16.23 (3.44) ms vs. gabazine: 11.40 (4.30) ms, P = 0.0068; for C, control: 8.23 (1.72) ms vs. gabazine: 5.5 (2.69) ms, P = 0.0147). D, representative graph of number of mIPSCs over time (one cell, young animal), according to the method described by Stell & Mody (2002). The frequency of mIPSC remains unaffected under partial GABAA blockade: the identical slope in number of events over time indicates that the frequency of all detected events in 200 nM gabazine (36 mIPSCs in total) is comparable to the 36 largest mIPSCs in control condition. E, gabazine does not affect mIPSC frequency (control: 0.51 (0.14) Hz vs. gabazine: 0.45 (0.18) Hz, P = 0.4342). Median values (inner line) are indicated in the box-plots and individual data points are averages from each cell. F, average mIPSC traces (the same 36 events recorded in control and gabazine conditions as in D) aligned for event rise onset. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: After recording spontaneous cortical activity in normal aCSF for 20 min, the perfusate was replaced with aCSF containing the GABAA antagonist gabazine (200 nm; SR95531 hydrobromide; Tocris Bioscience, Bristol, UK) and monitored for another 40–60 min.

Techniques: Control

Journal: The Journal of Physiology

Article Title: GABA_B‐ and GABA_A‐receptor‐mediated regulation of Up and Down states across development

doi: 10.1113/jp282736

Figure Lengend Snippet: Figure 10. Effect of 200 nM gabazine on phasic but not tonic currents A, representative traces from a pyramidal cell from a young animal. Bath application of 200 nM gabazine diminished mIPSCs, while 100 μM gabazine completely abolished them and evoked an outward shift in Ihold. B, all-points histograms generated for control (green), 200 nM gabazine (blue) and 100 μM gabazine (grey), corresponding to the coloured parts in A. C, after fitting Gaussian curves (see Methods), the mean value from each Gaussian fit is used to define the mean current in each condition. [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: After recording spontaneous cortical activity in normal aCSF for 20 min, the perfusate was replaced with aCSF containing the GABAA antagonist gabazine (200 nm; SR95531 hydrobromide; Tocris Bioscience, Bristol, UK) and monitored for another 40–60 min.

Techniques: Generated, Control

Journal: The Journal of Physiology

Article Title: GABA_B‐ and GABA_A‐receptor‐mediated regulation of Up and Down states across development

doi: 10.1113/jp282736

Figure Lengend Snippet: Figure 11. GABAA-mediated mIPSCs in pyramidal cells exhibit significant developmental modifications A, average traces of mIPSCs from voltage clamp recordings in young (red, average of 20 mIPSCs from one cell) and adult (blue, average of 20 mIPSCs from one cell) slices, respectively; coloured error bands correspond to SEM. B, raw traces at low temporal resolution of voltage-clamp recordings, illustrating the differences in mIPSC frequency. C, mIPSCs become larger, less frequent and with slower decays in adult (n (N) = 6 (6)) compared to young (n (N) = 6 (6)) animals. Only features with statistically significant age-difference are presented (for peak amplitude, young: −25.17 (12.64) pA vs. adult: −47.61 (10.55) pA, P = 0.0081; for frequency, young: 1.78 (0.34) Hz vs. adult: 0.87 (0.33) Hz, P < 0.0001; for decay τ w, young: 7.29 (0.81) ms vs. adult: 9.18 (1.07) ms, P = 0.0137). [Colour figure can be viewed at wileyonlinelibrary.com]

Article Snippet: After recording spontaneous cortical activity in normal aCSF for 20 min, the perfusate was replaced with aCSF containing the GABAA antagonist gabazine (200 nm; SR95531 hydrobromide; Tocris Bioscience, Bristol, UK) and monitored for another 40–60 min.

Techniques:

Journal: The European Journal of Neuroscience

Article Title: GABA B Receptors Mediate Intracellular Calcium Release in Astrocytes of the Prefrontal Cortex

doi: 10.1111/ejn.70187

Figure Lengend Snippet: GABA‐induced Ca 2+ transients in astrocytes are mediated by GABA B receptors. (A) GABA‐induced Ca 2+ transients are slightly enhanced in the presence of the GABA A receptor antagonist, gabazine (5 μM). Note that these experiments were performed in the presence of TTX, D‐APV, and NBQX to isolate direct GABAergic Ca 2+ responses in astrocytes with minimal neuronal influence. (B) Average amplitudes of GABA‐induced Ca 2+ transients normalized to the control application. (C) GABA‐induced Ca 2+ transients are strongly reduced in the presence of the GABA B antagonist, CGP 55845 (10 μM). Average amplitudes of GABA‐induced Ca 2+ transients normalized to the control application, showing a significant difference in the presence of the GABA B receptor inhibitor, CGP 55845. Statistical significance was determined by the Wilcoxon test for paired data sets. The error probability p was ** p < 0.01; *** p < 0.001.

Article Snippet: The reagents D‐2‐amino‐5‐phosphonovaleric acid (D‐APV; antagonist of NMDA receptors; #D‐145; working concentration: 100 μM), 2,3‐dioxo‐6‐nitro‐1,2,3,4‐tetrahydrobenzo[f]quinoxaline‐7‐sulfonamide (NBQX; antagonist of AMPA/kainate receptors; #N‐186; working concentration: 10 μM), tetrodotoxin (TTX; inhibiting voltage‐gated sodium channels; #T‐550; working concentration: 0.5 μM), and gabazine (antagonist of GABA A and Glycine receptors, #G‐215; working concentration: 5 μM) were purchased from Alomone Labs (Jerusalem, Israel).

Techniques: Control