Extended Data Fig. 3: Behavioral controls of ALAT KD in MB neurons (related to Fig. 2).
From: Glycolysis-derived alanine from glia fuels neuronal mitochondria for memory in Drosophila
a. ALAT knockdown (KD) in MB neurons impaired memory after single-cycle training in both females and males (females: n = 12, F2,33 = 5.13, P = 0.01; males: n = 12, F2,33 = 3.50, P = 0.04). ALAT KD in MB neurons impaired memory after spaced training in both females and males (females: n = 16, F2,45 = 13.14, P = 3.10−5; males: n = 14, F2,39 = 5.08, P = 0.01). b. Without induction of ALAT RNAi in MB neurons, memory after single-cycle training (n = 12, F2,33 = 0.92, P = 0.41) and spaced training (n = 12, F2,33 = 0.32, P = 0.73) was normal. c. The behavioral experiments were performed with a second RNAi against ALAT. ALAT knockdown in adult MB neurons impaired memory after single-cycle training (n = 10, F2,27 = 12.61, P = 0.00014) and spaced training (n = 11, F2,30 = 8.76, P = 0.001), but did not affect memory after single-cycle training followed by cold shock (n = 10, F2,27 = 0.09, P = 0.91) or massed training (n = 10, F2,27 = 1.90, P = 0.17). Without induction of ALAT RNAi in MB neurons, memory after single-cycle training (n = 12, F2,33 = 0.19, P = 0.83) and spaced training (n = 12, F2,27 = 0.77, P = 0.47) was normal. d. ALAT enzymatic activity in heads is decreased in flies mutant for ALAT (n = 8, t14 = 4.19, P = 0.0009). e. The ALAT mutation impaired memory after single-cycle training (n = 14 (control); n = 15 (mutant), t27 = 3.86, P = 0.0006) and spaced training (n = 12, t22 = 3.56, P = 0.002), but did not affect memory after single-cycle training followed by cold shock (n = 9, t16 = 0.18, P = 0.86) or massed training (n = 12, t22 = 0.86, P = 0.40). 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.
