(A) (left) Luminescence of DAN-aSP13 neurons expressing Lola-LUC reporter (MB315B-GAL4>UAS-FLP; Lola>stop>LUC) normalized to luminescence of the genetic control (UAS-FLP; Lola>stop> LUC). Mean …
(A) Expression pattern of MB315B-GAL4 line. (B) Luminescence of DAN-aSP13 neurons expressing Lola-LUC reporter (MB315B-GAL4>UAS-FLP; Lola>stop>-LUC) normalized to luminescence of the genetic control …
(A) Courtship indices (CIs) of the wild-type males trained in single pair assays with a mated female as indicated in Figure 1—figure supplement 1E and Table S1. (B) Courtship indices (CIs) of males …
(A) SIs of the wild-type males tested 24 hr after training for 6 hr with a mated female and sleep deprived at indicated time periods after training (dark purple bars). SI of the wild-type control …
(A) SIs of the wild-type males after 6 hr training with a mated female and sleep deprived during night at indicated time periods (dark purple bars). SI of the control males which were not deprived …
(A) Courtship indices (CIs) of the wild-type males trained in single pair assays with a mated female and sleep deprived as indicated in Figure 2A and Table S4. (B) Courtship indices (CIs) of males …
(A) (left) Expression pattern of 104y-GAL4 > UAS-Chrimson88-tdTomato with depicted dFB (magenta) and vFB (blue) layers for local activation with DMD. (right) Excitatory response of dFB (magenta) or …
(A) Expression pattern of the broad PAM-DANs driver (R58E02-LexA > LexAop-GCaMP6s). Manually defined DAN-γ5 (DAN-aSP13), DAN-γ4 and DAN-γ3 are depicted in white dashed circles. (B) (left) Expression …
(A) (left) Sleep profile of males (R23E10-GAL4 > UAS-TrpA1) upon activation of dFB neurons (red, n = 16) and control males with dFB neurons not activated (black, n = 16). Sleep time was plotted in …
(A) Three single FB neural cell types were manually traced in MCFO data: dFB (magenta), vFB (blue) and mFB (grey). Confocal images registered to standard brain of four FB lines in Figure 4 and …
(A) SIs of males of indicated genotypes tested 24 hr after 1 hr training with a mated female and activation at the specific time interval with CsChrimson (orange bars). SI of control males with …
(A) Courtship indices (CIs) of males trained in single pair assays with a mated female and with FB neurons activated as indicated in Figure 5A and Table S9. (B) Courtship indices (CIs) of males …
(A) The MBγ and DAN-aSP13s are repetitively activated during 6 hr training by the olfactory and behavioral cues presented by a mated female, respectively. (B) Males display an enhanced amount of …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Genetic reagent (D. melanogaster) | UAS-FLP.PEST; lola >> luc | DOI: 10.1073/pnas.1706608114 | Obtained from Michael Rosbash lab | |
Genetic reagent (D. melanogaster) | MB315B-GAL4 | DOI: 10.7554/eLife.04577 | Obtained from Gerry Rubin lab | |
Genetic reagent (D. melanogaster | UAS-myr::smGFP | DOI: 10.1534/genetics.110.119917 | BDSC ID: #32197 | Obtained from Gerry Rubin lab |
Genetic reagent (D. melanogaster) | Dopr1(attP) | DOI: 10.1038/nature11345 | Obtained from Barry Dickson lab | |
Genetic reagent (D. melanogaster) | VT005526-LexA | DOI: 10.1016/j.celrep.2015.05.037 | ||
Genetic reagent (D. melanogaster) | 104y-GAL4 | DOI: 10.1126/science.1202249 | FlyBase ID: FBti0072312 | Bloomington Stock Center |
Genetic reagent (D. melanogaster) | UAS-CsChrimson-mVenus | DOI: 10.1038/nmeth.2836 | BDSC ID: #55134, #55136 | Obtained from Vivek Jayaraman lab |
Genetic reagent (D. melanogaster) | LexAop2-CsChrimson-mVenus | DOI: 10.1038/nmeth.2836 | BDSC ID: #55139 | Obtained from Vivek Jayaraman lab |
Genetic reagent (D. melanogaster) | pBDP-GAL4 | DOI: 10.7554/eLife.04577 | Obtained from Gerry Rubin lab | |
Genetic reagent (D. melanogaster) | UAS-TrpA1 | DOI: 10.1101/gad.1278205 | ||
Genetic reagent (D. melanogaster) | LexAop-Shits | PMID: 12745632 | ||
Genetic reagent (D. melanogaster) | LexAop-myr::smGFP | DOI: 10.1534/genetics.110.119917 | BDSC ID: #32203 | Obtained from Gerry Rubin lab |
Genetic reagent (D. melanogaster) | UAS-GCaMP6s | DOI: 10.1073/pnas.1703090115 | Obtained from David Anderson lab | |
Genetic reagent (D. melanogaster) | R58E02-LexA | DOI: 10.1073/pnas.0803697105 | BDSC ID: #52740 | Obtained from Gerry Rubin lab |
Genetic reagent (D. melanogaster | UAS-Chrimson88-tdTomato | DOI: 10.1073/pnas.1703090115 | Obtained from David Anderson lab | |
Genetic reagent (D. melanogaster) | LexAop2-Syn21-opGCaMP6s | DOI: 10.1073/pnas.1703090115 | Obtained from David Anderson lab | |
Genetic reagent (D. melanogaster) | R23E10-GAL4 | DOI: 10.1073/pnas.0803697105 | BDSC ID: 49032 | Obtained from Gerry Rubin lab |
Genetic reagent (D. melanogaster) | VT036875-GAL4 | DOI: 10.1101/198648 | VDRC ID: 203402 | |
Genetic reagent (D. melanogaster) | R58E02-Gal80 | DOI: 10.1073/pnas.1703090115 | Flybase ID FBtp0097258 | Obtained from Glenn Turner lab |
Genetic reagent (D. melanogaster) | UAS-Shits | PMID: 12745632 | ||
Genetic reagent (D. melanogaster) | SS57264-GAL4 | This paper | generated by combining the VT045282-GAL4p65adz(attP40) and VT020829-ZpGAL4dbd(attP2) lines. | |
Chemical Compund, drug | Luciferin | GOLDBIO | Cat. LUCK | |
Chemical Compund, drug | Tetradotoxin | abcam | Cat. ab120054 | |
Software, algorithm | Permutation Test | PMID: 10355520 | ||
Software, algorithm | GraphPad Prism | GraphPad Prism (https://graphpad.com) | Version 7 | RRID:SCR_015807 |
Software, algorithm | FIJI | FIJI (https://imagej.net/Fij) | RRID:SCR_002285 |
Supplementary Tables.
Table S1. LTM does not depend on the circadian time of training Suppression indices (SIs) of naïve (train-) and experienced (train+) males of the indicated genotypes according to Figure 1—figure supplement 1E, tested in single-pair assays with mated females as trainers and testers. Courtship indices (CIs) are shown as median of n males and dispersion of the data as interquartile range (IQR). P values determined by permutation test for the null hypothesis that learning equals 0 (H0: SI = 0) or for the null hypothesis that experimental and control males learn equally well (H0: SI = SIc). Table S2. dopR1 mutant court mated females more than wild-type males Courtship indices (CIs) of naïve males of the indicated genotypes according to Figure 1—figure supplement 1G during 6 hr training with mated female are shown as median of n males and dispersion of the data as interquartile range (IQR). P values determined by permutation test for the null hypothesis that CIs of both groups are equal (H0: CIwt = CIDopR1). Table S3. dopR1 mutant males do not learn Suppression indices (SIs) of naïve (train-) and experienced (train+) males of the indicated genotypes according to Figure 1—figure supplement 1H, tested in single-pair assays with mated females as trainers and testers. Courtship indices (CIs) are shown as median of n males and dispersion of the data as interquartile range (IQR). P values determined by permutation test for the null hypothesis that learning equals 0 (H0: SI = 0) or for the null hypothesis that experimental and control males learn equally well (H0: SI = SIc). Table S4. Day-time sleep deprivation between 7–9 hr impairs LTM Suppression indices (SIs) of naïve (train-) and experienced (train+) males of the indicated genotypes, sleep deprived as denoted in Figure 2A, tested in single-pair assays with mated females as trainers and testers. Courtship indices (CIs) are shown as median of n males and dispersion of the data as interquartile range (IQR). P values determined by permutation test for the null hypothesis that learning equals 0 (H0: SI = 0) or for the null hypothesis that experimental and control males learn equally well (H0: SI = SIc). Table S5. Night-time sleep deprivation does not impair LTM Suppression indices (SIs) of naïve (train-) and experienced (train+) males of the indicated genotypes, sleep deprived as denoted in Figure 2-figure supplement A, tested in single-pair assays with mated females as trainers and testers. Courtship indices (CIs) are shown as median of n males and dispersion of the data as interquartile range (IQR). P values determined by permutation test for the null hypothesis that learning equals 0 (H0: SI = 0) or for the null hypothesis that experimental and control males learn equally well (H0: SI = SIc). Table S6. Silencing of DAN-aSP13 between 7–9 hr impairs LTM Suppression indices (SIs) of naïve (train-) and experienced (train+) males of the indicated genotypes with DAN-aSP13 silenced as denoted in Figure 2B, tested in single-pair assays with mated females as trainers and testers. Courtship indices (CIs) are shown as median of n males and dispersion of the data as interquartile range (IQR). P values determined by permutation test for the null hypothesis that learning equals 0 (H0: SI = 0) or for the null hypothesis that experimental and control males learn equally well (H0: SI = SIc). Table S7. Sleep induction between 5–7 hr consolidates STM to LTM Suppression indices (SIs) of naïve (train-) and experienced (train+) males of the indicated genotypes and sleep induced as denoted in Figure 2C, tested in single-pair assays with mated females as trainers and testers. Courtship indices (CIs) are shown as median of n males and dispersion of the data as interquartile range (IQR). P values determined by permutation test for the null hypothesis that learning equals 0 (H0: SI = 0) or for the null hypothesis that experimental and control males learn equally well (H0: SI = SIc). Table S8. DAN-aSP13 activation between 5–7 hr consolidates LTM Suppression indices (SIs) of naïve (train-) and experienced (train+) males of the indicated genotypes with DAN-aSP13 activated as denoted in Figure 2D, tested in single-pair assays with mated females as trainers and testers. Courtship indices (CIs) are shown as median of n males and dispersion of the data as interquartile range (IQR). P values determined by permutation test for the null hypothesis that learning equals 0 (H0: SI = 0) or for the null hypothesis that experimental and control males learn equally well (H0: SI = SIc). Table S9. Activation of vFB neurons between 5–7 hr consolidates LTM Suppression indices (SIs) of naïve (train-) and experienced (train+) males of the indicated genotypes with vFB activated as denoted in Figure 5A, tested in single-pair assays with mated females as trainers and testers. Courtship indices (CIs) are shown as median of n males and dispersion of the data as interquartile range (IQR). P values determined by permutation test for the null hypothesis that learning equals 0 (H0: SI = 0) or for the null hypothesis that experimental and control males learn equally well (H0: SI = SIc). Table S10. Silencing of vFB neurons between 7–10 hr impairs LTM Suppression indices (SIs) of naïve (train-) and experienced (train+) males of the indicated genotypes with vFB silenced as denoted in Figure 5B, tested in single-pair assays with mated females as trainers and testers. Courtship indices (CIs) are shown as median of n males and dispersion of the data as interquartile range (IQR). P values determined by permutation test for the null hypothesis that learning equals 0 (H0: SI = 0) or for the null hypothesis that experimental and control males learn equally well (H0: SI = SIc).
Fly genotypes.
Specific fly genotypes used in all main and supplementary figures.
Source code for analysis of calcium traces.