Fig. 2.

EtOH enhances evoked VTA GABA neuron IPSCs with block by α-conotoxins or knockdown of α6*-nAChRs. A Illustration showing the experimental framework wherein AAV-flex-mRuby2-α6miRNA was injected into the VTA 3 weeks prior and VTA GABA neurons were recorded in whole-cell, voltage-clamp mode in VGAT-Cre/GAD67-GFP mice. B Immunohistochemical panel showing that some VTA GABA neurons express α6*-nAChRs. An antibody against tyrosine hydroxylase (TH) was used along with the GAD67-GFP in these mice. Imaged using oil immersion 40 × objective (Olympus, UPlanFLN 1.30 numerical aperture). C Low-dose EtOH (5 mM) enhanced eIPSC amplitudes in VTA GABA neurons in VGAT-Cre/GAD67-GFP mice injected with scrambled α6-miRNA into the VTA, but not mice injected with α6-miRNA. There was no effect on paired-pulse ratio (50 ms), as previously reported [40]. Knockdown (KD) of α6*-nAChRs (D) or superfusion of MII (E) significantly reduced EtOH enhancement of VTA GABA neuron eIPSCs. Values in parentheses are n values. Asterisks ** indicate significance level p < 0.01