Journal: British Journal of Pharmacology
Article Title: Presence of ethanol‐sensitive and ethanol‐insensitive glycine receptors in the ventral tegmental area and prefrontal cortex in mice
Figure Lengend Snippet: Presence of glycine receptors in the mesolimbic circuit. (a) Western blot shows the presence of the α subunits (MW 48 kDa) of glycine receptors (α GlyR) in the VTA and PFC. (b) Graph represents the normalized signal of α GlyRs to Gβ. Data shown are means ± SEM; n = 5 (from duplicates). * P < 0.05, significantly different as indicated; unpaired Student's t ‐test. (c, d) Low magnification image of VTA and PFC (25×), respectively, in coronal brain slices from a GlyT2‐GFP mouse with TH (red) and anti‐GFP (green) signals in the VTA and MAP2 (red) and anti‐GFP (green) signals in the PFC. GlyT2‐positive projections appear in both regions. (e, f) High magnification image of the VTA and PFC (63×), respectively, in brain slices of a GlyT2‐GFP mouse showing GlyT2‐GFP (green) and GlyR α (magenta) signals. Confocal images of the VTA and PFC were replicated in three different mice
Article Snippet: Subsequently, the membranes were incubated with the following primary antibodies: anti‐glycine receptor pan α (1:500; rabbit monoclonal IgG; Cat No. 146008; Synaptic Systems, Germany; RRID:AB_2636914), anti‐glycine receptor β (1:200, rabbit polyclonal IgG, Cat No. AGR‐014, Alomone, RRID:AB_2340973) and anti‐Gβ (1:600, rabbit polyclonal IgG, Cat No. Sc‐378, Santa Cruz Biotechnology, RRID:AB_631542) for 1–2 h. After washes with 1× TBS and 0.1% Tween 20, membranes were incubated for 1 h with anti‐rabbit secondary antibodies conjugated to HRP (1:3000, goat polyclonal anti‐rabbit IgG‐HRP, Cat No. sc‐2004, Santa Cruz Biotechnology, RRID:AB_631746).
Techniques: Western Blot