Journal: International Journal of Molecular Sciences

Article Title: Impairment of Skeletal Muscle Contraction by Inhibitors of GABA Transporters

doi: 10.3390/ijms252312510

Figure Lengend Snippet: The absence of an inhibitory effect of β-alanine (1 mM) on contractions caused by indirect stimulation in the presence of a GABA B receptor blocker CGP 55845 (20 µM) in the Ringer–Krebs solution. The upper parts of the panels show native records of contractions obtained in individual experiments in control (black) and 20 min after chemical application (violet). The lower parts of the panels show the relative changes in the contraction force after β-alanine and CGP 55845 application relative to control (%) and time control. Data are presented as box plots with data overlap: diamonds—relative changes in individual experiments (n = 8 mice); horizontal lines—M ± SEM (boxes) and SD (whiskers). * p < 0.05 vs. time control (Mann–Whitney U test).

Article Snippet: The following primary antibodies were used in this study: GAT-1 GABA Transporter 1 (AGT-001, Alamone Labs, Jerusalem, Israel), GAT-2 GABA Transporter (AGT-002, Alamone Labs), and GABA T-3 Antibody (R-19) (sc-7669, Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Control, MANN-WHITNEY

Journal: International Journal of Molecular Sciences

Article Title: Impairment of Skeletal Muscle Contraction by Inhibitors of GABA Transporters

doi: 10.3390/ijms252312510

Figure Lengend Snippet: Micrographs from mouse diaphragm neuromuscular preparations processed for immunohistochemical analysis with specific antibodies against GABA transporters (GAT-1, GAT-2, and GAT-3; green). Labeling of nicotinic acetylcholine receptors (AChR) with tetramethylrhodamine-α-bungarotoxin was used as a marker of postsynaptic membrane of skeletal muscle fibers and synaptic region of neuromuscular preparation (red). Weak immunopositive reaction was revealed with antibodies against GAT-1; more pronounced immunopositive staining was detected when using antibodies against GAT-2. At the same time, immunohistochemical reaction with antibodies against GAT-3 was not detected. Scale bar = 10 μm.

Article Snippet: The following primary antibodies were used in this study: GAT-1 GABA Transporter 1 (AGT-001, Alamone Labs, Jerusalem, Israel), GAT-2 GABA Transporter (AGT-002, Alamone Labs), and GABA T-3 Antibody (R-19) (sc-7669, Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Immunohistochemical staining, Labeling, Marker, Membrane, Staining

Journal: Cell Research

Article Title: A GABAergic system in atrioventricular node pacemaker cells controls electrical conduction between the atria and ventricles

doi: 10.1038/s41422-024-00980-x

Figure Lengend Snippet: a Illustrator and workflow for the single-cell gene expression analysis of AVNPCs from Hcn4 CreERT2(+) ; Rosa26 TomRed+ mice. b Heatmap showing GABAergic system gene expression in single AVNPCs from Hcn4 CreERT2(+) ; Rosa26 TomRed+ mice. Rows indicate the AVNPC samples, and columns indicate the cycle threshold (Ct) values of GABAergic system genes. Ct values were scaled and the relative gene expression was demonstrated using color scales. A score of –1 (red) indicates a high expression level, and a score of 1 (blue) indicates a low level of expression. The data were obtained from 52 AVNPCs from 5 Hcn4 CreERT2(+) ; Rosa26 TomRed+ mice. c Immunofluorescence staining showing the expression and localization of GABA metabolic enzymes (GAD2, GABA-T, and SSADH), GABA A receptors (GABRA3, GABRB2, and GABRG2), GABA transporters (vGAT and GAT-1) in single mouse AVNPCs. Scale bar, 10 μm.

Article Snippet: The following primary antibodies were used in immunofluorescence experiments: anti-GABA antibody (5A9) (Abcam, ab86186, 1:100 for cells); anti-GABRA3 antibody (Alomone, AGA-003, 1:200 for cells, 1:50 for heart slices); anti-GABRB2 antibody (Invitrogen, PA5-64331, 1:100 for cells, 1:20 for heart slices); anti-GABRG2 antibody (Alomone, AGA-005, 1:100 for cells, 1:50 for heart slices); anti-GAT-1 antibody (Synaptic Systems, 274102, 1:200 for cells, 1:50 for heart slices); anti-vGAT antibody (Alomone, AGT-005, 1:100 for cells, 1:50 for heart slices); anti-GAD2 antibody (Santa Cruz Biotechnology, sc-377145, 1:100 for cells, 1:50 for heart slices); anti-GABA-T antibody (Abcam, ab216465, 1:100 for cells, 1:50 for heart slices); anti-SSADH antibody (Santa Cruz Biotechnology, sc-390754, 1:100 for cells, 1:50 for heart slices); anti-CX43 antibody (Cell Signaling Technology, 3512, 1:50 for heart slices); anti-HCN4 antibody produced in mouse (Sigma, SAB5200035, 1:100 for cells, 1:50 for heart slices); anti-HCN4 antibody produced in rat (Sigma, MABN1871, 1:100 for cells, 1:50 for heart slices); and anti-CAST antibody (Invitrogen, PA5-87352, 1:100 for cells, 1:50 for heart slices).

Techniques: Expressing, Immunofluorescence, Staining