Figure 3
Detection of responsive neurons from spaced and massed trained flies. (A) LTM conditioning consists in a repeated presentation of an odor paired with electric shock. (B) An illustrative sample from the unpaired control group. The volumetric reconstruction on the left shows the mCherry nuclei signal together with GCaMP activity during the OCT response. On the right, the GCaMP signal is shown together with the detected nuclei. The plot shows the normalized signal from the individual neurons. Light gray spheres and lines represent neurons that did not respond, and the dark gray color indicates neurons that responded. Three neurons are highlighted in pink, yellow and blue (with the corresponding signal of the same color) to show that the signal of individual neurons can be monitored in different parts of the mushroom body. (C) An illustrative sample from the paired conditioning group using the same visualization as in panel B. (D) Comparison of the numbers of responsive neurons per fly between the paired 5x spaced conditioning to form LTM (n = 29) and the unpaired control (n = 27) groups (Mann-Whitney two-sided test p-value: 0.001098) as well as between the paired 5x massed conditioning to form LT-ARM (n = 14) and the unpaired control (n = 16) groups (Mann-Whitney two-sided test p-value: 0.97894). Results show that the number of responsive neurons increases after paired conditioning but not after massed paired conditioning. (E) Comparison of the mean intensity of responsive neurons per fly at stimulus time in the same flies and conditions as in panel D. Statistical tests could not reject the null hypothesis of an equal mean distribution of intensity (t-test p-value 5x spaced paired vs unpaired p-value: 0.882074 and t-test p-value 5x massed paired vs unpaired p-value: 0.5291736).
