(A) Representative maximum projection confocal stacks of MB018B > trans-Tangorevealed that β`2mp glutamatergic MBON is postsynaptic to α`2 cholinergic neuron (B) Representative maximum projection confocal stacks of MB058B > trans-Tangosuggests that that α`2 cholinergic MBON is not postsynaptic α2α`2 DAN (C) Thermogenetic inactivation of α`2 cholinergic neurons during acquisition did not affect the expression of alcohol-associated preference F(2,63)=2.18, p=0.12. (D) Inactivation of α`2 cholinergic neurons during retrieval significantly reduced preference F(2,116)=19.46, p=5.17×10^−08. (E) Similarly, thermogenetic inactivation of α2α`2 DANs during acquisition did not affect the expression of alcohol-associated preference F(2,85)=0.202, p=0.817. (F) Inactivation during retrieval significantly reduced preference F(2,54)=5.103, p=0.009. (G) Updated model of circuits responsible for the expression of alcohol-associated preference during retrieval. Scale bar = 50 μm.