Neuronal reactivation during post-learning sleep consolidates long-term memory in Drosophila

  1. Ugur Dag
  2. Zhengchang Lei
  3. Jasmine Q Le
  4. Allan Wong
  5. Daniel Bushey
  6. Krystyna Keleman  Is a corresponding author
  1. Howard Hughes Medical Institute, United States
6 figures, 1 table and 4 additional files

Figures

Figure 1 with 2 supplements
DAN-aSP13 neurons are activated during sleep.

(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 …

https://doi.org/10.7554/eLife.42786.003
Figure 1—figure supplement 1
Learning sufficient to induce LTM leads to the enhencement of the post-learning sleep and activation of DAN-aSP13 in the specific time window.

(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 …

https://doi.org/10.7554/eLife.42786.004
Figure 1—figure supplement 2
Courtship Indices (CIs) of males that had undrgone treatment according to Figure 1—figure supplement 1E and H.

(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 …

https://doi.org/10.7554/eLife.42786.005
Figure 2 with 2 supplements
Sleep after learning is necessary and sufficient for LTM consolidation.

(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 …

https://doi.org/10.7554/eLife.42786.006
Figure 2—figure supplement 1
Activation of FB neurons induces sleep.

(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 …

https://doi.org/10.7554/eLife.42786.007
Figure 2—figure supplement 2
Courtship Indices (CIs) of males that had undergone treatment according to Figure 2 and Figure 2—figure supplement 1A.

(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 …

https://doi.org/10.7554/eLife.42786.008
Figure 3 with 1 supplement
FB neurons provide an excitatory input to DAN-aSP13.

(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 …

https://doi.org/10.7554/eLife.42786.009
Figure 3—figure supplement 3
Activation of the mFB has no effect on DAN-aSP13 activity.

(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 …

https://doi.org/10.7554/eLife.42786.010
Figure 4 with 1 supplement
Sleep promoting vFB neurons activate DAN-aSP13.

(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 …

https://doi.org/10.7554/eLife.42786.011
Figure 4—figure supplement 1
Specific class of vFB neurons acutely induces sleep.

(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 …

https://doi.org/10.7554/eLife.42786.012
Figure 5 with 1 supplement
Sleep promoting vFB neurons are sufficient and necessary for LTM consolidation.

(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 …

https://doi.org/10.7554/eLife.42786.013
Figure 5—figure supplement 5
Courtship Indices (CIs) of males that had undergone treatment according to Figure 5A and B.

(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 …

https://doi.org/10.7554/eLife.42786.014
Post-learning activation of DAN-aSP13 neurons mediates LTM consolidation.

(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 …

https://doi.org/10.7554/eLife.42786.015

Tables

Key resources table
Reagent type
(species) or
resource
DesignationSource or
reference
IdentifiersAdditional
information
Genetic reagent
(D. melanogaster)
UAS-FLP.PEST; lola >> lucDOI: 10.1073/pnas.1706608114Obtained from Michael
Rosbash lab
Genetic reagent
(D. melanogaster)
MB315B-GAL4DOI: 10.7554/eLife.04577Obtained from
Gerry Rubin lab
Genetic reagent
(D. melanogaster
UAS-myr::smGFPDOI: 10.1534/genetics.110.119917BDSC ID:
#32197
Obtained from
Gerry Rubin lab
Genetic reagent
(D. melanogaster)
Dopr1(attP)DOI: 10.1038/nature11345Obtained from
Barry Dickson lab
Genetic reagent
(D. melanogaster)
VT005526-LexADOI: 10.1016/j.celrep.2015.05.037
Genetic reagent
(D. melanogaster)
104y-GAL4DOI: 10.1126/science.1202249FlyBase ID:
FBti0072312
Bloomington
Stock Center
Genetic reagent
(D. melanogaster)
UAS-CsChrimson-mVenusDOI: 10.1038/nmeth.2836BDSC ID:
#55134, #55136
Obtained from
Vivek Jayaraman lab
Genetic reagent
(D. melanogaster)
LexAop2-CsChrimson-mVenusDOI: 10.1038/nmeth.2836BDSC ID:
#55139
Obtained from Vivek
Jayaraman lab
Genetic reagent
(D. melanogaster)
pBDP-GAL4DOI: 10.7554/eLife.04577Obtained from
Gerry Rubin lab
Genetic reagent
(D. melanogaster)
UAS-TrpA1DOI: 10.1101/gad.1278205
Genetic reagent
(D. melanogaster)
LexAop-ShitsPMID: 12745632
Genetic reagent
(D. melanogaster)
LexAop-myr::smGFPDOI: 10.1534/genetics.110.119917BDSC ID:
#32203
Obtained from
Gerry Rubin lab
Genetic reagent
(D. melanogaster)
UAS-GCaMP6sDOI: 10.1073/pnas.1703090115Obtained from
David Anderson lab
Genetic reagent
(D. melanogaster)
R58E02-LexADOI: 10.1073/pnas.0803697105BDSC ID:
#52740
Obtained from
Gerry Rubin lab
Genetic reagent
(D. melanogaster
UAS-Chrimson88-tdTomatoDOI: 10.1073/pnas.1703090115Obtained from David
Anderson lab
Genetic reagent
(D. melanogaster)
LexAop2-Syn21-opGCaMP6sDOI: 10.1073/pnas.1703090115Obtained from David
Anderson lab

Genetic reagent
(D. melanogaster)
R23E10-GAL4DOI: 10.1073/pnas.0803697105BDSC ID:
49032
Obtained from
Gerry Rubin lab
Genetic reagent
(D. melanogaster)
VT036875-GAL4DOI: 10.1101/198648VDRC ID:
203402
Genetic reagent
(D. melanogaster)
R58E02-Gal80DOI: 10.1073/pnas.1703090115Flybase ID
FBtp0097258
Obtained from Glenn
Turner lab
Genetic reagent
(D. melanogaster)
UAS-ShitsPMID: 12745632
Genetic reagent
(D. melanogaster)
SS57264-GAL4This papergenerated by combining
the VT045282-GAL4p65adz(attP40)
and VT020829-ZpGAL4dbd(attP2) lines.
Chemical
Compund, drug
LuciferinGOLDBIOCat. LUCK
Chemical
Compund, drug
TetradotoxinabcamCat. ab120054
Software, algorithmPermutation TestPMID: 10355520
Software, algorithmGraphPad PrismGraphPad Prism
(https://graphpad.com)
Version 7RRID:SCR_015807
Software, algorithmFIJIFIJI
(https://imagej.net/Fij)
RRID:SCR_002285

Additional files

Supplementary file 1

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).

https://doi.org/10.7554/eLife.42786.016
Supplementary file 2

Fly genotypes.

Specific fly genotypes used in all main and supplementary figures.

https://doi.org/10.7554/eLife.42786.017
Source code 1

Source code for analysis of calcium traces.

https://doi.org/10.7554/eLife.42786.018
Transparent reporting form
https://doi.org/10.7554/eLife.42786.019

Download links