Fig. 4.

CeA-delivered pHSVsiLTLR4a selectively inhibits binge alcohol drinking. (A) Binge alcohol responding in P rats on an FR-4 schedule during the presurgery (5 d), and after pHSVsiLTLR4 infusion into the CeA. The asterisk represents significance for pHSVsiLTLR4 (n = 10) compared with pHSVsiNC (n = 8) and the presurgery control using the Tukey and Dunnett's test, respectively, following significant group [F_(1,10) = 15.46, P < 0.003] and session [F_{(10, 10)} = 3.42, P < 0.032] effects. (B) Binge sucrose responding in P rats (n = 5) on an FR-4 schedule during the presurgery (5 d) and after pHSVsiLTLR4 infusion in the CeA. Except for the initial postsurgery day, pHSVsiLTLR4a did not significantly alter binge sucrose responding (P > 0.05). (C) Cohorts of binged P rats were microinfused with PBS, pHSVsiNC, or pHSVsiLTLR4a into the CeA and micropunches collected 72 h or 15 d after infusion. Protein extracts were then immunoblotted with antibodies to α2, TLR4, or GAPDH control and blots stripped between antibodies and expressed as densitometric units ± SEM pHSVsiLTLR4a inhibited TLR4 at day 3, but not day 15 relative to PBS and pHSVsiNC (P > 0.05); pHSVsiLTLR4a also failed to alter the α2 protein (P > 0.05). (D) Group of EGFP⁺ neurons near one of the pHSVsiLTLR4a injection sites in the CeA. (Scale bar, 25 μm.) (E) Binge alcohol responding in P rats on an FR-4 schedule during the presurgery (5 d), and after pHSVsiLTLR4 (n = 7) and pHSVsiNC (n = 8) infused into the VP; pHSVsiLTLR4a did not alter binge alcohol responding in the VP [F_{(1, 10)} = 2.78, P > 0.05]. (See SI Materials and Methods for blood alcohol levels and additional statistical details for A–C and E.)