Extended Data Fig. 6: Behavioral controls of PFK KD in cortex glia, and pyruvate imaging in PFK KD (related to Fig. 4).
From: Glycolysis-derived alanine from glia fuels neuronal mitochondria for memory in Drosophila
a. PFK knockdown (KD) in cortex glia impaired memory in both females and males after single-cycle (females: n = 16, F2,44 = 6.89, P = 0.003; males: n = 17 (white bar); n = 19 (gray bar); n = 18 (pink bar), F2,51 = 3.22, P = 0.048) and spaced training (females: n = 14, F2,39 = 5.04, P = 0.01; males: n = 14, F2,39 = 4.95, P = 0.01). b. Without induction of PFK RNAi, memory after single-cycle (n = 12, F2,33 = 0.18, P = 0.84) and spaced training (n = 10, F2,27 = 0.24, P = 0.79) were normal. c. Behavioral results were confirmed with a second RNAi against PFK. Although no significant effect on memory after single-cycle training was revealed (total: n = 13, F2,36 = 1.60, P = 0.22; with cold shock: n = 13, F2,36 = 4.56, P = 0.02), subtracting the memory after cold shock from the total memory, which measures only MTM, revealed an MTM impairment (n = 13, F2,36 = 5.23, P = 0.01). LTM was also impaired (n = 20, F2,57 = 5.10, P = 0.009), but not memory after massed training (n = 14, F2,39 = 3.05, P = 0.06). Without induction of PFK RNAi, all memories were normal (single-cycle: n = 12, F2,33 = 0.60, P = 0.55; single-cycle followed by cold shock: n = 12, F2,33 = 0.41, P = 0.67; MTM: n = 12, F2,33 = 0.38, P = 0.69; spaced training: n = 18, F2,51 = 0.35, P = 0.70). d. The increased mitochondrial pyruvate flux elicited by single-cycle training in MB neuron somata (n = 12, t22 = 3.36, P = 0.003) was impaired by PFK knockdown in adult cortex glia (n = 12, t22 = 1.88, P = 0.07). All data are presented as mean ± SEM. Asterisks (*P < 0.05; **P < 0.01; ***P < 0.001; ns: not significant, P > 0.05) illustrate the significance level of two-sided t-test, or of the least significant pairwise comparison following 1-way ANOVA.
