Extended Data Fig. 6: , related to Fig. 4: Control experiments for mitochondrial network remodeling facilitates ATP production in the soma of MB neurons and improves memory after massed training.
From: Neuronal fatty acid oxidation fuels memory after intensive learning in Drosophila
Extended Data Figure 6a. The mitochondria volume distribution is shown for each ROI of all unpaired control flies used in this study, where each dot represents the volume of one single identified mitochondrion. The means of the quartile limits from all control flies subjected to the massed unpaired protocol (that is tub-Gal80ts; VT30559 > UAS-mtDsRed flies subjected to the unpaired 5x massed protocol in Fig. 4a-b) were used to define the 4 mitochondria categories. b-c. related to Fig. 4c. b: Mean traces of ATP consumption of MB neurons of flies subjected to either massed associative training (green) or the unpaired protocol (black). c: Mean traces of ATP consumption of MB neurons KD for Drp1 of flies subjected to either massed associative training (red) or the unpaired protocol (black). d related to Fig. 4d: Mean traces of ATP consumption of MB neurons of flies subjected to unpaired protocol (red: Drp1 KD flies, blue: Drp1, MTPa KD flies, black: No RNAi flies). e. When Drp1 RNAi expression was not induced, memory after massed training was normal (n = 15, F2,42 = 2.01, P = 0.147). Drp1 KD in adult MB neurons impaired memory after spaced training (n = 17, F2,48 = 3.68, P = 0.0325). When Drp1 RNAi expression was not induced, memory after spaced training was normal (n = 19, F2,54 = 2.25, P = 0.116). f. A second non-overlapping RNAi targeting Drp1 (Drp1 RNAi GD10456) was used to confirm the specific increase in memory after massed training. Inhibition of Drp1 expression in adult MB neurons using this RNAi increased memory performance after massed training (n = 18, F2,51 = 5.53, P = 0.0067), whereas memory after spaced training was impaired (n = 18, F2,51 = 4.38, P = 0.0176). Non-induced flies showed normal memory performance after either massed training (n = 19,18,19, F2,53 = 1.33, P = 0.274) or spaced training (n = 17, F2,48 = 2.01, P = 0.146). g. Tango11 KD in adult MB neurons increased memory performance after massed training (n = 19, F2,54 = 6.93, P = 0.0021), whereas memory after spaced training was impaired (n = 21, F2,60 = 6.05, P = 0.0040). Non-induced flies showed normal memory performance after either massed training (n = 16,17,17, F2,47 = 1.49, P = 0.236) or spaced training (n = 12, F2,33 = 2.28, P = 0.118). h. The increase of memory performance observed after massed training in flies KD for Drp1 in adult MB neurons was preserved when ketone body metabolism was impaired (Drp1, ACAT1 KD) (n = 12, F4,55 = 10.63, P = 0.000002, the p-value of the relevant pairwise comparisons are indicated). The red statistical marker shows the comparison between Drp1 KD and ACAT1, Drp1 KD flies. Data on barplots are expressed as mean ± s.e.m. with dots as individual values, and analyzed by ANOVA with post hoc testing by Newman–Keuls pairwise comparisons test (e-h). Genotype sample sizes are listed in the legend in order of bar appearance. Significance level of the Newman–Keuls pairwise comparison between the genotype of interest and the genotypic controls (e-h) following one-way ANOVA: *P < 0.05, **P < 0.01, ***P < 0.001, NS: not significant.
