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. 2020 Jun 4;9:e48730. doi: 10.7554/eLife.48730

Figure 5. A microcircuit within the vertical lobe is important for alcohol-associated preference.

(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.

Figure 5.

Figure 5—figure supplement 1. Decreasing expression of dopamine receptors in the α`2 MBON did not affect alcohol-associated preference.

Figure 5—figure supplement 1.

RNAi knockdown of either D2Rs (A) or D1Rs (B) within MBON α`2 using the MB018B split-GAL4 driver did not significantly affect alcohol-associated preference. One-way ANOVA was used to compare mean and variance with Tukey Posthoc compared to experimental (A) (F(2,57)=0.113, p=0.894 and (B) F(2,37)=1.997, p=0.15, respectively).
Figure 5—figure supplement 2. Temperature controls for lines that showed decreases in retrieval of alcohol-associated preference at a restrictive temperature.

Figure 5—figure supplement 2.

Flies were trained and tested at permissive temperatures (20°C) and exhibited normal alcohol associated preference for pair odor cues under these conditions.