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

Figure 4. MBON β`2mp glutamatergic neuron is postsynaptic to β`2a PAM DANs and important for memory consolidation.

(A) Representative maximum projection confocal stacks of MB109B > transTango. (B) trans-Tango reveal the α`/β` MB lobe (i) and MBON β`2mp neurons (ii) as postsynaptic to β`2a DANs. (C) Thermogenetic inactivation of MBON β`2mp during consolidation using MB002B significantly increased alcohol reward preference F(2,54) = 9.287, p=0.0003. Thermogenetic inactivation of β`2mp during consolidation using MB074C significantly increased alcohol reward preference relative to UAS controls F(2,71) = 3.51, p=0.04. (D) Knockdown of D2R in MBON β`2mp using MB002B significantly decreased alcohol-associated preference F(2,63)=12.77, p=2.22×10^−05. Knockdown of D2R in MBON β`2mp using MB074C significantly decreased alcohol-associated preference relative to GAL4 controls F(2,71)=3.51, p=0.04. One-way ANOVA with Tukey Posthoc was used to compare mean and variance. Bar graphs illustrate mean +/- standard error of the mean. *p<0.05 **p<0.01 (f) Circuits responsible for encoding alcohol-associated preference during retrieval. Scale bar = 50 μm.

Figure 4.

Figure 4—figure supplement 1. RNAi knockdown of dopamine receptors in β`2 MBONs in alcohol-associated preference.

Figure 4—figure supplement 1.

(A) or D1Rs (B) within MBON β2β`2a using MB399B split-GAL4 driver did not significantly affect alcohol-associated preference (F(2,65)=0.032, p=0.968 and F(2,41)=0.223, p=0.801, respectively). (C) RNAi knockdown of D1Rs within MBON β`2mp using the MB074C split-GAL4 driver did not significantly affect alcohol-associated preference F(2,79)=0.123, p=0.884. One-way ANOVA was used to compare mean and variance with Tukey Posthoc compared to experimental.
Figure 4—figure supplement 2. Temperature controls for lines that showed decreases in retrieval of alcohol-associated preference at a restrictive temperature.

Figure 4—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.