Journal: Frontiers in Molecular Neuroscience

Article Title: Dissection of signaling pathways regulating TrkB-dependent gephyrin clustering

doi: 10.3389/fnmol.2024.1480820

Figure Lengend Snippet: Chemically induced long-term potentiation (chem iLTP) at inhibitory synapses relies on TrkB signaling. (A, B) Representative images of TrkB WT expressing primary neurons after treatment with sham solution (A) or NMDA+CNQX (B) for chem iLTP induction. (A’,B’) Gphn staining. (A”,B”) VGat staining. (A”’,B”’) Gphn/VGat. Scale bars: 5 μm. Overview images and individual datapoints are shown in . (C–G) Analysis of gephyrin clustering in neurons expressing TrkB WT and mutants after chem-iLTP or sham treatment as indicated. Gray dots: sham treatment. Orange dots: chem iLTP. Numerical data are means ± SEM, obtained from three independent experiments.Values are normalized to the corresponding untreated group. Statistical significance was assessed by two-way ANOVA with Tukey’s multiple comparison test (* p < 0.05, ** p < 0.01, *** p < 0.001; non-significant comparisons are not noted). For detailed statistical values refer to . (C) Total somatic gephyrin cluster densities: TrkB WT sham, 1.00 ± 0.05; TrkB WT chem iLTP, 1.14 ± 0.05; TrkB SHC- sham, 1.00 ± 0.05; TrkB SHC- chem iLTP, 1.05 ± 0.05; TrkB PLC- sham, 1.00 ± 0.04; TrkB PLC- sham chem iLTP, 1.12 ± 0.05; TrkB KD, 1.00 ± 0.04; TrkB KD chem iLTP, 1.05 ± 0.05; n = 80–90 somata. (D) Synaptic somatic gephyrin cluster densities. TrkB WT sham, 1.00 ± 0.05; TrkB WT chem iLTP, 1.24 ± 0.07; TrkB SHC- sham, 1.00 ± 0.05; TrkB SHC- chem iLTP, 0.93 ± 0.06; TrkB PLC- sham, 1.00 ± 0.05; TrkB PLC- chem iLTP, 1.03 ± 0.05; TrkB KD, 1.00 ± 0.05; TrkB KD chem iLTP, 0.84 ± 0.05; n = 80–90 somata. (E) Mean gephyrin cluster size. TrkB WT sham, 1.00 ± 0.05; TrkB WT chem iLTP, 1.17 ± 0.05; TrkB SHC- sham, 1.00 ± 0.04; TrkB SHC chem iLTP, 0.94 ± 0.04; TrkB PLC- sham, 1.00 ± 0.04, TrkB PLC- chem iLTP, 1.22 ± 0.05; TrkB KD, 1.00 ± 0.04; TrkB KD chem iLTP, 0.98 ± 0.04; n = 78–90 somata. (F) Mean gephyrin cluster size. CTR sham, 1.00 ± 0.03; CTR chem iLTP, 1.11 ± 0.04; PD98059 sham, 1.00 ± 0.03; PD98059 chem iLTP, 0.90 ± 0.03; KN-62 sham, 1.00 ± 0.03; KN-62 chem iLTP, 1.05 ± 0.03; Cyclotraxin B sham, 1.00 ± 0.03; Cyclotraxin B chem iLTP, 0.90 ± 0.03; n = 82–89 somata. Corresponding microscopic images are shown in .

Article Snippet: At DIV14, the cells were washed and treated with 145 mM NaCl, 2 mM KCl, 2 mM CaCl, 2 mM MgCl, 10 mM Glucose, 10 mM HEPES ( ) supplemented with 20 μM NMDA and 10 μM CNQX (MedChemExpress, Monmouth Junction, NJ) for 2 min, followed by a recovery period in the treatment solution for 18 min. To inhibit MEK1, CaMKII or TrkB, neurons were treated with either 50 μM PD98059, 3 μM KN-62, 1 μM Cyclotraxin B (all from MedChemExpress, Monmouth Junction, NJ) or DMSO (CTR) for 30 min in culture medium before induction of chem iLTP as described.

Techniques: Expressing, Staining, Comparison

Journal: eNeuro

Article Title: Hippocampus-Dependent Goal Localization by Head-Fixed Mice in Virtual Reality

doi: 10.1523/ENEURO.0369-16.2017

Figure Lengend Snippet: Statistical table

Article Snippet: The water-soluble competitive AMPA/kainate-type glutamate receptor antagonist CNQX disodium salt (Tocris) was dissolved in cortex buffer (123 mM NaCl, 5 mM KCl, 10 mM glucose, 2 mM CaCl 2 , 2 mM MgCl 2 , and 10 mM HEPES, pH 7.4) at a concentration of 1 mg/ml (3.45 mM).

Techniques: Control, Mann-Whitney U-Test

Journal: Scientific Reports

Article Title: Light-evoked hyperpolarization and silencing of neurons by conjugated polymers

doi: 10.1038/srep22718

Figure Lengend Snippet: ( a ) Analysis of the distribution of neurons grown on either glass:P3HT or glass and responding with a decreased, increased or stable firing rate during the light pulse (Glass:P3HT vs Glass: decreasing 55.56% vs 0.00%; stable 37.04% vs 100.00%, increasing 7.40% vs 0.00%). The percentage of cell showing a decrease during the pulse on glass:P3HT is statistically significant with respect to those on glass (p < 0.0001, Fisher’s test). For neurons on glass:P3HT showing a decrease in the firing rate during the light pulse, the distribution of the response after the pulse is also described as part of the whole (decreasing 13.33%; stable 20.00%, increasing 66.67%). ( b ) Mean (±sem) hyperpolarization amplitude recorded in neurons grown on glass:P3HT (grey and black bars) or glass (white bar) in the presence of the synaptic blockers AP5/CNQX/PTX (glass:P3HT, −5.74 ± 0.71 mV, n = 27; glass:P3HT with blockers, −5.62 ± 1.55 mV, n = 14; Glass, −1.08 ± 0.10 mV, n = 14; Glass:P3HT vs Glass ****p < 0.0001, Glass:P3HT Blockers vs Glass ****p < 0.0001, Glass:P3HT vs Glass:P3HT Blockers p > 0.9999; Kruskal-Wallis test with Dunn’s multiple comparison test). ( c ) Linear correlation between the hyperpolarization amplitude during illumination and the firing rate after illumination in individual neurons grown on glass:P3HT (Pearson’s correlation coefficient r = 0.69, n = 27; p < 0.001). ( d ) Amplitude of the hyperpolarization response to 15 consecutive stimulations (green bars) performed at 0.1 Hz. Potential traces were averaged over the recorded cells on glass:P3HT ( n = 27). Left panel: full traces for sweeps 1, 2 and 15. Right panel: average hyperpolarization amplitude of cells grown onto glass:P3HT over the repeated stimulations (sweeps 1–15).

Article Snippet: To block excitatory and inhibitory synaptic transmission, recordings were performed in the presence of AP5 (50 μM), CNQX (10 μM) and PTX (100 μM) all from Tocris (USA).

Techniques: Comparison