Extended Data Fig. 2: , related to Fig. 1 Control experiments for ATP imaging experiments.
From: Neuronal fatty acid oxidation fuels memory after intensive learning in Drosophila
Extended Data Fig. 2a. Applying 5 mM of sodium azide (dashed line) resulted in a fast decrease in the FRET ratio of the ATP FRET sensor AT1.03NL expressed in adult MB neurons, whereas the FRET ratio of AT1.03RK (a nonfunctional version of the probe that is ATP insensitive) did not change (mean trace ± s.e.m.). The slope of the FRET ratio was measured between 90% and 30% of the baseline level (plateau reached after sodium azide application corresponding to the lower limit of the FRET sensor detection). b. related to Fig. 1e: Mean traces of ATP consumption of MB neurons of flies subjected to either associative massed training (green) or the unpaired protocol (black). c. XY-plot of individual fly datapoints of baseline ATP level and the corresponding slopes of all control flies (No RNAi and subjected to unpaired massed protocol) showing no correlation between ATP baseline level and the rate of ATP decrease (the slope) triggered by sodium azide application (Pearson correlation: n = 49, R² = 0.06, p = 0.09). d. related to Fig. 1e: Mean traces of ATP consumption of MB neurons KD for MTPα of flies subjected to either massed associative training (blue) or the unpaired protocol (black). e. In the β’2 subdomain of the β’ lobes (Region of recording of MB neurons outline in white dotted line, scale bar: 15 μm), massed associative training (green) triggered an increase of ATP consumption compared to flies subjected to the unpaired protocol (black; n = 8,10, t16 = 2.49, P = 0.0243), a representative trace of recording is presented for each condition f. Mean traces of ATP consumption recorded in the β’2 subdomain of MB neurons of flies subjected to either massed associative training (green) or the unpaired (black). g. In the soma of MB neurons, no difference in ATP consumption was observed after spaced training (purple: associative, black: non-associative protocol, n = 11, t20 = 1.06, P = 0.304), a representative trace of recording is presented for each condition. h. Mean traces of ATP consumption recorded in the soma of MB neurons of flies subjected to either spaced associative training (purple) or the non-associative spaced protocol (black). Data on barplots are expressed as mean ± s.e.m. with dots as individual values, and analyzed by two-sided unpaired t-test (e,g). Genotype sample sizes are listed in the legend in order of bar appearance. Significance level of a two-sided unpaired t-test (e-g): *P < 0.05, **P < 0.01, ***P < 0.001, NS: not significant.
