Figure 2.

Behavioral response in the T-maze after targeted cell ablation of Mz317 cells

(A) Temporal control of expression directed by the Mz317-Gal4 driver using the Gal80^ts element for producing viable fly adults with cell ablation. The Gal80^ts element blocks the action of the Gal4 element at low temperatures (19°C), but at high temperatures (30°C), the Gal80^ts element is inactivated, allowing the action of Gal4 to direct the expression of the UAS-gene insert. In these panels, we tested the presence of Mz317 cells in the third antennal segment after cell ablation for 4 days in the adult stage with diphtheria toxin (DTl). At 19°C, the GFP reporter present in the UAS-GFP insert is not expressed. At 30°C, we can see GFP expression in cells that do not express the diphtheria toxin, but Mz317 cells do not appear in the Mz317-Gal4/UAS-DTl; Gal80ts/UAS-mcD8:GFP antenna because the expression of DTl kills these cells. Scale bar, 40 μm

(B) The breeding protocol indicates the applied temperatures during different time periods. Cell ablation is controlled by temperature blocking of the restrictive action of Gal80^ts at 30°C. Below, the dose-response curve of control flies compared with the experimental flies that underwent Mz317 cell ablation for 4 days in the adult stage (for Canton-S/UAS-DTl, N₁₀₋₂ = 22, N_10−1.5 = 22, N₁₀₋₁ = 27; for Mz317-Gal4/UAS-DTI; Gal80^ts, N₁₀−2 = 20, N₁₀−1.5 = 20, N₁₀−1 = 25). Each data point represents the mean +/- SEM.

(C) Above, experimental procedure indicating the breeding treatment applied to both genotypes. Below, olfactory preference response to the odorant ethyl acetate at the 10⁻¹ concentration in the T-maze. Only the flies with MZ317 cell ablation differed significantly from the other 3 groups (for Canton-S/UAS-DTl, N_CONTROL = 15, N_EXP = 14; for Mz317-Gal4/UAS-DTl; Gal80^ts, N_CONTROL = 15, N_EXP = 13). In all cases, each data point represents the mean ± SEM of 20 replicate tests with 20 flies each.

∗∗∗ = p < 0.001, ∗ = p < 0.05.