Figure 4. Memory retention overnight.

(A) Testing and training protocols. Except where indicated, larvae were tested, trained immediately after testing, tested again, then placed on food overnight and tested the following day. For extinction experiments, larvae were trained three times, and then exposed to 18 cycles of alternating CO₂ and air either immediately following training or prior to testing the next day. (B,C,D) Probability of choosing CO₂ containing channel (top) and fraction of larvae in trained group according to double Gaussian model fit (bottom) prior to training, immediately following training, and the next day. When the center bar is missing, larvae were not tested immediately following training but instead removed immediately to food. M Nx = massed training, N repetitions, S 10x = spaced training 10 total pairings, RP = reverse paired (see Figure 1C), No Train = no training. Larvae in (B,C) were DANi1>CsChrimson. Larvae in (D) were DANi1>hs-dCREB2-b;CsChrimson. Larvae were raised on food containing ATR, except for ATR+/CXM-, ATR+/CXM+ larvae who were fed ATR supplemented yeast paste (without/with cycloheximide) for 4 hr prior to initial testing. For reverse-paired (RP) and no training schemes, see Figure 1B. * p<0.05, ** p<0.01, *** p<0.001.