Fig. 4: SFSs integrate MBONs-γ2α’1 and MBONs-β’2mp inputs onto vFBs.
From: A neural circuit linking learning and sleep in Drosophila long-term memory
a Neural skeletons derived from the FIB-SEM Drosophila brain volume. b Confocal image of SFSs expression pattern extracted from the Multi Color Flip-Out (MCFO) of the VT033828-GAL4 line. Scale bar = 50 μm. (Details in the Methods). c, d Traces of mean GCaMP6s responses (ΔF/F) (±SEM) in SFSs upon optogenetic activation (red bar) of MBONs-γ2α’1 (c, n = 9, P = 0.048), MBONs-β’2mp (d, n = 5, P = 0.0002). P value is for H0 ΔF/F = 0, two-sided Student T test. e. Traces of mean GCaMP6s responses (ΔF/F) (±SEM) in vFBs upon SFSs focal activation (red bar). n = 6, P = 0.0075 H0 ΔF/F = 0, two-sided Student T test. f. Mean ΔSleep (exp-nai) in the 1–3 h time period after training (TP1-3) upon SFSs silencing. n = 39 flies per group, P1 = 0.0046, P2 = 0.1208, P3 = 1.1e−4, P4 = 0.0445 for H0 ΔSleep = 0, two-sided Wilcoxon Signed Rank test, and P12 = 0.0091, P34 = 0.2711 for H0 ΔSleep20 °C = ΔSleep30 °C, two-sided Wilcoxon Rank Sum test. g LTM, shown as SI [%], after SFSs silencing in TP1-3. n1 = 52 and 52, n2 = 61 and 64, n3 = 62 and 51, n4 = 65 and 62 for naïve and trained group respectively, P1 = 1.0e−5, P2 = 0.4976, P3 = 0.0028, P4 = 2.0e−4 for H0 SI = 0, and P12 = 0.0115, P34 = 0.7954 for H0 SIexp = SIctrl; two-sided Permutation test. h Mean calcium traces (±SEM) in SFSs, measured for 1 min at multiple time points spanning TP1-3. i Mean calcium levels in TP1-3 in h. n0 h = 28, n1 h = 17, n2 h = 18, n4 h = 19, n6 h = 26, P1 h = 0.3431, P2 h = 0.6126, P4 h = 0.0312, P6 h = 0.0131 for H0 Fnai = Fexp, P = 0.0466 for H0 F2 h = F4 h, and P = 0.8225 for H0 F4 h = F6 h, two-sided Student T test. j Model of the circuit mechanism to generate post-learning sleep for LTM consolidation. Solid lines indicate monosynaptic connections. Full genotypes and data analysis details in Supplementary Tables 1–3. Source data are provided as a Source Data file. ns P > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001. n represents independent fly samples with assays repeated at least 3 times (c, d, e, f, g and i). Box plots represent median and IQR, whiskers extend to lower and upper adjacent values and red crosses for outliers (f and i).
