Article Figures & Data
Figures
- Fig. 1.
Intact acute, but absence of sensitized ethanol (EtOH) locomotor response in AC1KO mice (A–C), n = 6–7/group, and WT mice pretreated with the selective inhibitor NB001 (D–F), 10 mg/kg, n = 8–9/group. Average locomotor activity (A and D) and time course following acute (B and E) or challenge (C and F) EtOH (2.0 g/kg) after repeated home cage EtOH administration (2.5 g/kg, 10 injections). Data reported as percentage change relative to respective saline-only controls. WT EtOH-treated mice displayed an acute locomotor response relative to saline-only WT controls, which was significantly enhanced following sensitization. AC1KO and NB001-treated WT mice demonstrated a comparable acute locomotor response to WT EtOH controls, but failed to display a further potentiation in this response following sensitization. Locomotor activity on the acute and challenge day was unaltered by pretreatment with NB001 alone (NB001 + saline). Significance determined by two-way RM ANOVA and Sidak’s post hoc test, *P ≤ 0.05, compared with respective saline-treated controls; ¥P < 0.05, compared with WT + acute EtOH or saline + acute EtOH group; #P < 0.05, compared with WT + challenge EtOH or saline + acute (E) or challenge (D, F) EtOH group.
- Fig. 2.
Repeated ethanol (EtOH) selectively enhanced GluN2B-NMDAR phosphorylation in the DMS in WT, but not AC1KO (KO) mice or mice pretreated with NB001 (n = 3–5/group). Averaged data (left) and representative immunoblots compiled from groups of images from different parts of the same gel (right) are depicted for DMS (A), DLS (B), or NAc (C) lysates prepared following repeated saline (sal) or EtOH treatment and probed for total and pTyr-1472-GluN2B. (A) A significant increase in pGluN2B levels in the DMS of WT mice was observed following repeated EtOH treatment relative to saline-treated controls, which was absent in AC1KO and NB001 pretreated mice. (B) Repeated ethanol treatment decreased levels of pGluN2B in the DLS equivalently across WT, AC1KO, and NB001-treated mice. (C) No changes in pGluN2B levels in the NAc were observed in response to ethanol or absence of AC1 signaling. Total GluN2B levels were unaffected by AC1 status or ethanol treatment in all three regions. Significance determined by two-way ANOVA and Sidak’s post hoc test, *P < 0.05, compared with saline-treated controls; #P < 0.05, compared with WT EtOH.
- Fig. 3.
Ethanol (EtOH) upregulation of NMDAR-mediated transmission is absent in AC1KO (KO) mice. (A) Representative traces of NMDAR-mediated eEPSCs and (B) average NMDAR-mediated eEPSC amplitude (± S.E.M.) in WT and AC1KO mice repeatedly treated with saline (sal) or EtOH. EtOH produced an increase in NMDAR-mediated eEPSC amplitude only in WT mice (two-way ANOVA and Sidak’s post hoc test, **P < 0.01). (C) Average NMDAR-mediated eEPSC amplitude before (BL) and after (+Ro) bath application of the GluN2B selective antagonist Ro 25-6981 (0.5 µM) in WT mice. EtOH-induced upregulation of NMDAR-mediated transmission involved an increased contribution of GluN2B-containing NMDARs (two-way RM ANOVA and Sidak’s post hoc test, *P < 0.05). (D) Average central location of medium spiny neuron recordings was 1.10 mm from bregma (± 0.5 mm).
