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. 2020 Aug 17;30(16):3200–3211.e8. doi: 10.1016/j.cub.2020.05.077

Figure 4.

Figure 4

Aversive Memory Extinction and Sugar Learning Require Different Subsets of PAM-γ5 DANs

(A) Brain from a VT006202-GAL4; UAS-GFP fly labels all 20 PAM-γ5 DANs and possibly some other PAM DANs (black). Three commissures are visible (asterisk). Brain co-stained with nc82 antibody (MB is outlined). Scale bar, 20 μm.

(B) MB315C-GAL4; UAS-GFP specifically labels 8 PAM-γ5 DANs per hemisphere that cross the midline in the lower commissure.

(C) 0804-GAL4 labels 5 PAM-γ5 DANs per hemisphere, previously “γ5 narrow,” that occupy the lower commissure. Scale bar, 20 μm.

(D) 0104-GAL4; UAS-GFP labels PAM-γ5 DANs, previously named “γ5 broad,” that cross the midline in the upper and middle commissures. 0104 also labels some other PAM DANs [9].

(E) Table summarizing DAN expression in GAL4 lines used for behavior, modified from [9]. R48B04GAL80 refines the 0104-GAL4 expression [12], shown in the Data S1.

(F) Aversive olfactory memory extinction. Schematic: experimental timeline and temperature shift protocol. Blocking neuron output during odor re-exposure impaired memory extinction in R58E02-, VT006202-, and 0804-GAL4; UAS-Shits1, but not in MB315C- or 0104-GAL4 ± GAL80; UAS-Shits1 flies. Bars show mean ± SEM. Asterisks denote p < 0.035 (wild-type) and p < 0.0176 (0104); one-way ANOVA with Tukey’s post hoc test, n = 10–12.

(G) Olfactory learning with sucrose reinforcement. Schematic: experimental timeline and temperature shift protocol. Blocking neuron output during training impaired 30 min appetitive memory in R58E02-, VT006202-, and 0104-GAL4 ± GAL80; UAS-Shits1, but not MB315C- or 0804-GAL4; UAS-Shits1 flies. Bars show mean ± SEM. Asterisks denote p < 0.0003 (R58E02), p < 0.0004 (0104), p < 0.0018 (VT006202), and p < 0.0010 (R48B04GAL80; 0104-GAL4); one-way ANOVA with Dunnett’s post hoc test, n = 10.

(H) Schematic of input pathways to PAM-γ5 and PAM-β′2a DANs and subtype innervation of the γ5 compartment. Colors correspond to clusters defined in Figure 1. Open circles represent 2 γ5 DANs that were identified but not further analyzed.

See also Figure S4, Data S1, and Video S4.