gaba b receptor agonist  (Tocris)

Journal: bioRxiv

Article Title: Reduced GABA B receptor activation and post activation depression of proprioceptive afferents after spinal cord injury

doi: 10.64898/2026.01.21.700955

Figure Lengend Snippet: A) Schematic of dorsal root (DR) and ventral root (VR) recordings in response to low-intensity stimulation of Ia afferents in adjacent DR (pink) and optogenetic activation of dorsal (light blue) and ventral (dark blue) projecting GAD2 neurons by blue light in GAD2//ChR2 mice. Dorsal GAD2 neurons innervate GABA A receptors (purple) near Ia nodes to evoke PAD and ventral GAD2 neurons innervate GABA B receptors (green) at Ia terminal to produce presynaptic inhibition, and GABA A receptors on motoneuron to produce an IPSP. Excitatory interneurons between Ia afferent and GAD2 neurons are coloured in lavender. B) DR response to a focused 10 ms light pulse [447nm laser, 0.7 mW/mm 2 , 1.5x light threshold (LT) to evoke PAD or motoneuron responses] applied to dorsal and ventral location of the whole sacral spinal cord (“s” indicates PAD-evoked spike). C) Top: DR response to the same ventral light pulse alone. Middle: DR-evoked EPSP without (pink) and with (blue) prior ventral light pulse, Bottom: DR-evoked EPSP without (blue) and with (green) CGP55845. D) Change in EPSP from prior ventral light pulse without and with CGP55845 (CGP) and in GAD2- mice. E-F) Amplitude of IPSP (E) and PAD (H) from ventral or dorsal light with and without gabazine. G) Dorsal (top) and ventral (bottom) root responses with (blue) and without (pink) PAD evoked from light pulse to dorsal spinal cord with DR-evoked EPSPs triggered during (left) and after (right) PAD. H) Change in EPSP when evoked during and after dorsal light PAD, and during PAD with and without CGP55845 and gabazine (Gbz) (447 nm laser, 1.1xLT). I) Same as in G but with stronger dorsal light pulse to produce PAD-evoked spikes in afferents (green) compared to no-spiking PAD (light blue, 1.2xLT). J) Difference in change in EPSP with PAD-evoked spikes from dorsal light pulse in control and SCI mice and in response to ventral light pulse. In D-J, * indicates p < 0.05 for paired and unpaired comparisons to the control variable; in H-J, + indicates p < 0.001 when the variable is compared to a 0% change or difference (see Supplemental Table 2A for mean (SD) values in D to J and results of statistical comparisons used).

Article Snippet: CGP55845 (abbreviated CGP in figures), a GABA B receptor antagonist (0.3 μM), gabazine, a GABA A receptor antagonist (50 μM) and baclofen, a GABA B receptor agonist (1 μM) (all from Tocris), were added to the nACSF to examine their effects on post-activation depression and Ia hyperpolarization.

Techniques: Activation Assay, Inhibition, Control

Journal: bioRxiv

Article Title: Reduced GABA B receptor activation and post activation depression of proprioceptive afferents after spinal cord injury

doi: 10.64898/2026.01.21.700955

Figure Lengend Snippet: A) Schematic of dorsal root (DR) stimulation and ventral root (VR) recordings to produce post-activation depression of Ia-EPSPs. B) Evoked EPSPs from repetitive DR stimulation every 60ms in GAD2//Arch3 mice without (pink trace) and with (green trace) application of light to silence GAD2 neurons (532nm laser, 5 mW/mm 2 ). Amplitude of 4 th EPSP (M4) is compared to first EPSP (M1). C) Percent change of 4 th EPSP compared to 1 st EPSP in GAD2//Arch3 mice (n = 8 VR recordings from 3 mice) without (white) and with (green) silencing of GAD2 neurons with light. Percentage change of EPSPs in wild type mice (n = 8 VR recordings in 3 mice) without (white) and with (green) GABA B receptor blockade with CGP55845. D-E) Repetitive stimulation of Ia-EPSP from dorsal root stimulation alone (pink traces) and when combined with light pulses to ventral spinal cord (blue traces; 447nm laser, 0.7 mW/mm 2 , 1.5xLT)) in GAD2//ChR2 mice without (D) and during (E) GABA B receptor blockade using CGP55845. F) Repetitive light stimulation of ventral root without (top) and with (bottom) CGP55845 before evoking a test EPSP with DR stimulation. Light blue trace (top) shows small EPSP from PAD-evoked spike. G) Change in Ia-EPSP size in response to repetitive dorsal root, ventral light or combined stimulation with and without CGP845 to indicate GABA B receptor effect on post-activation depression. H) Baseline membrane potential of motoneuron in response to repetitive dorsal root stimulation (pink) or ventral light stimulation in GAD2//ChR2 mice without, and in presence of, CGP55845 and gabazine (GBZ). In C and G, * indicates p < 0.05 for paired Student t-tests and in H, + indicates p < 0.05 from a 0 mV change. Supplemental Table 2B contains mean (SD) values in C, G and H and results of statistical comparisons used.

Article Snippet: CGP55845 (abbreviated CGP in figures), a GABA B receptor antagonist (0.3 μM), gabazine, a GABA A receptor antagonist (50 μM) and baclofen, a GABA B receptor agonist (1 μM) (all from Tocris), were added to the nACSF to examine their effects on post-activation depression and Ia hyperpolarization.

Techniques: Activation Assay, Membrane

Journal: bioRxiv

Article Title: Reduced GABA B receptor activation and post activation depression of proprioceptive afferents after spinal cord injury

doi: 10.64898/2026.01.21.700955

Figure Lengend Snippet: A) Schematic of immunolabelling of Ia afferents and GAD2+ interneuron terminal and associated GABA B receptors and VGLUT1+ afferent terminals. B) Left: Mean intensity of GABA B receptors on the Ia terminal of SCI mice as a % of the intensity in intact mice, with pairs of injured and control spinal cord sections placed on the same slide for labelling. Right: Mean intensity of GABA B receptors on the motoneuron of SCI mice as a % of the intensity on the motoneurons of control mice. C) Mean intensity of GABA B receptors on the Ia terminal as a % of the intensity on the motoneuron for SCI and control mice. D) Mean number of terminals with GABA B receptors as a % of all identified Ia terminals. E-F) Transverse section of spinal cords from intact ( E ) and chronic SCI ( F ) mice whose afferents were labeled with tdTOM (red; AAV9-tdTom), GABA B receptors in green and GAD2 interneurons in blue in GAD2//ChR2-EYFP mice (top row). Lower left panels showing enlarged images of afferents (2nd row), GABA B receptors (3rd row), GAD2 neurons (4th row) and merged picture from all three (5 th row). Right panels: expanded view of Ia afferent terminal for data on left with additional labelling of Ia terminal with VGLUT1 in 4th and 5 th row. GABA B receptors computed to be in the volume of the terminal are rendered yellow. N = 5 mice per group. * p < 0.05 for unpaired Student’s t-tests. Mean (SD) and statistical results in Supplemental Table 2C.

Article Snippet: CGP55845 (abbreviated CGP in figures), a GABA B receptor antagonist (0.3 μM), gabazine, a GABA A receptor antagonist (50 μM) and baclofen, a GABA B receptor agonist (1 μM) (all from Tocris), were added to the nACSF to examine their effects on post-activation depression and Ia hyperpolarization.

Techniques: Control, Labeling

Journal: Oncology Letters

Article Title: In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists

doi: 10.3892/ol.2025.15103

Figure Lengend Snippet: γ-aminobutyric acid B receptor activation affects the numbers of migrated cells. (A) MDA-MB-231 cells (2×10 4 ) were seeded into the upper chambers of the Transwell plates. Following 24 h baclofen treatment at different concentrations (0, 25, 50, 100 and 200 µM), the numbers of cells that had migrated were counted, and images were captured under the microscope at ×20 magnification. (B) Data are shown as the mean ± standard error of the mean from three independent experiments. Statistical analysis was performed using one-way ANOVA with Dunnett's post hoc test. ***P<0.001 and ****P<0001 compared with the untreated control. Bac, baclofen.

Article Snippet: The GABA B receptor agonist baclofen (Tocris Bioscience) was dissolved in water to form a stock solution (concentration, 10 mM) and stored at −20°C until use.

Techniques: Activation Assay, Microscopy, Control

Journal: Oncology Letters

Article Title: In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists

doi: 10.3892/ol.2025.15103

Figure Lengend Snippet: γ-aminobutyric acid B receptor activation affects the numbers of invaded cells. (A) Representative images of invaded MDA-MB-231 cells following baclofen treatment are shown. The upper chambers of the Transwell plates were pre-coated with 70 µl Matrigel for 1 h. MDA-MB-231 cells (5×10 4 ) were seeded in each upper chamber and treated with baclofen at various concentrations (0, 25, 50, 100 or 200 µM) for 48 h. Images were captured at ×20 magnification. (B) Data are presented as the mean ± standard error of the mean from three independent experiments. Statistical analysis was performed using one-way ANOVA with Dunnett's post hoc test. ***P<0.001 and ****P<0001 compared with the untreated control. Bac, baclofen.

Article Snippet: The GABA B receptor agonist baclofen (Tocris Bioscience) was dissolved in water to form a stock solution (concentration, 10 mM) and stored at −20°C until use.

Techniques: Activation Assay, Control

Journal: Oncology Letters

Article Title: In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists

doi: 10.3892/ol.2025.15103

Figure Lengend Snippet: γ-aminobutyric acid B receptor activation inhibits cell survival and proliferation. (A) MDA-MB-231 cells (200 per well) were seeded in 24-well plates and treated with various concentrations of baclofen (0, 25, 50, 100 and 200 µM) for 10 days. Images were captured using a digital camera. (B) Data are shown as the mean ± standard error of the mean from three independent experiments. Statistical analysis was performed using one-way ANOVA with Dunnett's post hoc test. **P<0.01 and ****P<0001 compared with the untreated control. Bac, baclofen; ns, not significant.

Article Snippet: The GABA B receptor agonist baclofen (Tocris Bioscience) was dissolved in water to form a stock solution (concentration, 10 mM) and stored at −20°C until use.

Techniques: Activation Assay, Control

Journal: Oncology Letters

Article Title: In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists

doi: 10.3892/ol.2025.15103

Figure Lengend Snippet: γ-aminobutyric acid B receptor activation exerts no effect on cell viability. MDA-MB-231 cells (2×10 4 ) were seeded into each upper chamber of a 24-well plate, and the cells were treated with baclofen at different concentrations (0, 25, 50, 100 and 200 µM) for 3 days. DMSO at a concentration of 0.01% was used as a control and the numbers of cells that survived were counted. The viability of the cells was found not to be affected following treatment with baclofen. The MTT assays were repeated three times, and each experiment yielded similar results. Data are shown as the mean ± standard error of the mean from three independent experiments. One-way ANOVA with Dunnett's post hoc test was performed to calculate statistical analysis of the mean compared with the untreated control. ns, not significant.

Article Snippet: The GABA B receptor agonist baclofen (Tocris Bioscience) was dissolved in water to form a stock solution (concentration, 10 mM) and stored at −20°C until use.

Techniques: Activation Assay, Concentration Assay, Control

Journal: Oncology Letters

Article Title: In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists

doi: 10.3892/ol.2025.15103

Figure Lengend Snippet: γ-aminobutyric acid B receptor activation modulates epithelial-mesenchymal transition markers in MDA-MB-231 breast cancer cells. (A) Representative immunoblots showing the expression of β-catenin and vimentin in MDA-MB-231 cells treated with various concentrations of baclofen. Following 24 h of serum starvation, cells were exposed to baclofen (0, 25, 50, 100, 200 µM) for 24 h. GAPDH served as a loading control. (B) Quantification of the expression of β-catenin relative to GAPDH is shown. Baclofen treatment induced dose-dependent increases in β-catenin levels, with significant upregulation observed at concentrations of 50, 100 and 200 µM. (C) Quantification of the expression of vimentin relative to GAPDH is shown. A significant decrease in vimentin expression was observed at a baclofen concentration of 100 and 200 µM. Data represent the mean ± standard error of the mean. from at least three independent experiments. Protein band intensities were quantified using ImageJ software. Statistical analysis was performed using one-way ANOVA with Dunnett's post hoc test for comparisons with the control group. *P<0.05, **P<0.01 and ***P<0.001. ns, not significant; C, control, Bac, baclofen.

Article Snippet: The GABA B receptor agonist baclofen (Tocris Bioscience) was dissolved in water to form a stock solution (concentration, 10 mM) and stored at −20°C until use.

Techniques: Activation Assay, Western Blot, Expressing, Control, Concentration Assay, Software

Journal: Oncology Letters

Article Title: In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists

doi: 10.3892/ol.2025.15103

Figure Lengend Snippet: Effects of γ-aminobutyric acid B receptor activation on the P13K/Akt and Ras/Raf/MAPK signaling pathways. (A) After 24 h of treatment with different concentrations of baclofen (25, 50, 100 and 200 µM), the phosphorylation levels of Akt and ERK1/2 were found not to have been significantly altered. (B) Quantification of phospho-Akt/Akt. (C) Quantification of phospho-ERK/ERK. The experiments were repeated three times with similar results. Protein quantification was performed using ImageJ software. Statistical analysis was performed using one-way ANOVA with Dunnett's post hoc test for comparisons with the control group. The data are shown as the mean ± standard error of the mean. ERK, extracellular signal-regulated kinase; PI3K, phosphatidylinositol 3-kinase; Akt, protein kinase B; MAPK, mitogen-activated protein kinase; ns, not significant; Bac, baclofen; phospho, phosphorylated.

Article Snippet: The GABA B receptor agonist baclofen (Tocris Bioscience) was dissolved in water to form a stock solution (concentration, 10 mM) and stored at −20°C until use.

Techniques: Activation Assay, Protein-Protein interactions, Phospho-proteomics, Software, Control

Journal: Oncology Letters

Article Title: In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists In vitro and in silico investigation of the critical role of GABA B receptor activation in human breast cancer and its natural agonists

doi: 10.3892/ol.2025.15103

Figure Lengend Snippet: Chemical structures, TC similarity scores, binding scores and hydrogen bond interactions of baclofen and the three docked compounds with the γ-aminobutyric acid B receptor active site.

Article Snippet: The GABA B receptor agonist baclofen (Tocris Bioscience) was dissolved in water to form a stock solution (concentration, 10 mM) and stored at −20°C until use.

Techniques: Binding Assay