gaba a receptor blocker sr 95531  (Alomone Labs)

Journal: EMBO Reports

Article Title: Synaptotagmin-11 facilitates assembly of a presynaptic signaling complex in post-Golgi cargo vesicles

doi: 10.1038/s44319-024-00147-0

Figure Lengend Snippet: ( A ) Representative traces of sEPSCs from Syt11 +/+ (black) and Syt11 −/− (blue) cultured primary hippocampal neurons recorded in the presence of gabazine (10 µM) at DIV15-19. ( B ) Cumulative probability distribution of sEPSC inter-event intervals in Syt11 +/+ and Syt11 −/− neurons. The sEPSC frequency (inset) was significantly reduced in Syt11 −/− compared to Syt11 +/+ neurons ( Syt11 +/+ : 9.92 ± 1.25 Hz vs Syt11 −/− : 6.03 ± 0.56 Hz). ( C ) Cumulative probability distribution of sEPSC amplitudes in Syt11 +/+ and Syt11 −/− neurons. The average sEPSC amplitude (inset) was not significantly different between the genotypes ( Syt11 +/+ : 22.06 ± 2.06 pA vs Syt11 −/− : 23.55 ± 1.73 pA). ( D ) Representative traces of sEPSCs recorded from a Syt11 +/+ (top) and Syt11 −/− (bottom) neuron in the presence of gabazine (10 µM) before (control, black/red) and after application of baclofen (100 μM, blue). ( E ) Cumulative probability distributions of sEPSC inter-event intervals from Syt11 +/+ (top) and Syt11 −/− (bottom) neurons. In both genotypes, the sEPSC frequency (insets) was significantly reduced in the presence of baclofen (bac) compared to control (con). Upper inset: Syt11 +/+ neurons (con: 9.92 ± 1.25 Hz vs bac: 2.66 ± 0.62 Hz). Lower inset: Syt11 −/− neurons (con: 6.03 ± 0.56 Hz vs bac: 2.32 ± 0.29 Hz). ( F ) Cumulative probability distributions of sEPSC amplitudes from Syt11 +/+ (top) and Syt11 −/− (bottom) neurons. In both genotypes, the average amplitude (insets) was significantly decreased in the presence of baclofen (bac) compared to control (con). Upper inset: Syt11 +/+ neurons (con: 22.06 ± 2.06 pA vs bac: 15.23 ± 1.18 pA). Lower inset: Syt11 -/ neurons (con: 23.55 ± 1.73 pA vs bac: 14.06 ± 1.10 pA). ( G ) Number of sEPSCs plotted against sEPSC amplitudes in Syt11 +/+ neurons in the presence and absence (con) of baclofen (left). Activation of GBRs significantly inhibits small amplitude (<10 pA) and large amplitude (>10 pA) events (right). ( H ) Number of sEPSCs plotted against sEPSC amplitudes in Syt11 −/− neurons in the presence and absence (con) of baclofen (left). Activation of GBRs significantly inhibits large amplitude (>10 pA) but not small amplitude (<10 pA) events. Data information: Data are presented as mean ± SEM. Statistical significance was determined by Mann–Whitney U test ( B ), unpaired Student’s t -test ( C ), paired Student’s t -test ( E , upper inset and F ) or Wilcoxon matched-pairs signed-rank test ( E , lower inset; G and H ). ns not significant; ** p < 0.01, *** p < 0.001, **** p < 0.0001. Syt11 +/+ , n = 14 neurons; Syt11 −/− , n = 19 neurons from 6 preparations. .

Article Snippet: Spontaneous glutamatergic PSCs were recorded in the presence of the GABA A receptor blocker gabazine (SR 95531 hydrobromide, 10 μM) (Cat# 1262, Tocris; 10 mM stock dissolved in water). (RS)-Baclofen (100 μM) (Cat# 0417, Tocris; 100 mM stock dissolved in 1eq NaOH) was used to activate GABA B receptors.

Techniques: Cell Culture, Control, Activation Assay, MANN-WHITNEY

Journal: EMBO Reports

Article Title: Synaptotagmin-11 facilitates assembly of a presynaptic signaling complex in post-Golgi cargo vesicles

doi: 10.1038/s44319-024-00147-0

Figure Lengend Snippet: ( A ) Representative traces of sEPSCs recorded from a Syt11 +/+ (top) and Syt11 −/− (bottom) neuron in the presence of gabazine (10 µM) before (control, black/red) and after application of CGP54626 (4 μM, yellow). ( B ) Cumulative probability distributions of sEPSC inter-event intervals from Syt11 +/+ (top) and Syt11 −/− (bottom) neurons recorded as in ( A ). In both genotypes, the sEPSC frequency (insets) was not significantly different in the presence of CGP54626 (CGP) compared to control (con). Upper inset: Syt11 +/+ neurons (con: 8.16 ± 2.46 Hz vs CGP: 7.07 ± 1.83 Hz). Lower inset: Syt11 −/− neurons (con: 5.74 ± 1.32 Hz vs CGP: 6.31 ± 1.67 Hz). n = 5 neurons per genotype from 3 preparations. ( C ) Summary bar graph showing the sEPSC frequency of Syt11 +/+ (left) and Syt11 −/− (right) neurons in the presence of gabazine (10 µM) before (control, black/red) and after application of baclofen (100 µM, blue) and baclofen + CGP54626 (4 µM, yellow). Syt11 +/+ , n = 11 neurons; Syt11 −/− , n = 16 neurons from 6 preparations. Data information: Data are presented as mean ± SEM. Statistical significance was determined by Wilcoxon matched-pairs signed-rank test ( B ) or Friedman test and Dunn’s multiple comparisons ( C ). ns not significant; **** p < 0.0001.

Article Snippet: Spontaneous glutamatergic PSCs were recorded in the presence of the GABA A receptor blocker gabazine (SR 95531 hydrobromide, 10 μM) (Cat# 1262, Tocris; 10 mM stock dissolved in water). (RS)-Baclofen (100 μM) (Cat# 0417, Tocris; 100 mM stock dissolved in 1eq NaOH) was used to activate GABA B receptors.

Techniques: Control

Journal: EMBO Reports

Article Title: Synaptotagmin-11 facilitates assembly of a presynaptic signaling complex in post-Golgi cargo vesicles

doi: 10.1038/s44319-024-00147-0

Figure Lengend Snippet: ( A ) Representative traces of sEPSCs from Syt11 +/+ and Syt11 −/− hippocampal neurons in culture recorded at DIV15-19 in the presence of gabazine (10 µM) before (control, black/red) and after application of ω-conotoxin (1 µM, blue), ω-conotoxin + ω-agatoxin (500 nM, yellow) and ω-conotoxin + ω-agatoxin + TTX (1 µM, magenta). ( B ) Cumulative probability distributions of sEPSC inter-event intervals of Syt11 +/+ and Syt11 −/− neurons recorded as in ( A ). ( C ) Summary bar graph depicting the sEPSC frequency of Syt11 +/+ and Syt11 −/− neurons recorded as in ( A ). In both genotypes, the frequency of sEPSCs was significantly reduced by the application of ω-conotoxin, ω-conotoxin + ω-agatoxin, and ω-conotoxin + ω-agatoxin + TTX. ( D ) Summary bar graph depicting the percentage inhibition of sEPSC frequency by ω-conotoxin, ω-agatoxin, and TTX in cultured hippocampal neurons of Syt11 +/+ (black) and Syt11 −/− mice (red). Inhibition by ω-conotoxin (blocking Cav2.2 channels) is significantly reduced in Syt11 −/− compared to Syt11 +/+ neurons ( Syt11 +/+ : 60.55 ± 4.42% vs Syt11 −/− : 31.90 ± 7.80%). The ω-agatoxin-sensitive (blocking Cav2.1 channels) and TTX-sensitive components of inhibition, as well as the TTX-insensitive component (mEPSCs) show no significant difference between genotypes. ( E ) The combined ω-agatoxin- and TTX-sensitive component of inhibition is significantly increased in Syt11 −/− compared to Syt11 +/+ neurons ( Syt11 +/+ : 29.33 ± 4.64% vs Syt11 −/− : 47.78 ± 7.37%). Data information: Data are presented as mean ± SEM. Statistical significance was determined by Wilcoxon matched-pairs signed-rank test with Bonferroni correction for multiple comparison ( C ) or Mann–Whitney U test ( D and E ). ns not significant; * p < 0.05, ** p < 0.01. n = 9–11 neurons per genotype from 4 preparations. .

Article Snippet: Spontaneous glutamatergic PSCs were recorded in the presence of the GABA A receptor blocker gabazine (SR 95531 hydrobromide, 10 μM) (Cat# 1262, Tocris; 10 mM stock dissolved in water). (RS)-Baclofen (100 μM) (Cat# 0417, Tocris; 100 mM stock dissolved in 1eq NaOH) was used to activate GABA B receptors.

Techniques: Control, Inhibition, Cell Culture, Blocking Assay, Comparison, MANN-WHITNEY