Presynaptic inhibition of dopamine neurons controls optimistic bias

  1. Nobuhiro Yamagata  Is a corresponding author
  2. Takahiro Ezaki
  3. Takahiro Takahashi
  4. Hongyang Wu
  5. Hiromu Tanimoto  Is a corresponding author
  1. Graduate School of Life Sciences, Tohoku University, Japan
  2. Research Center for Advanced Science and Technology, The University of Tokyo, Japan
8 figures, 1 table and 2 additional files

Figures

Figure 1 with 1 supplement
Gamma-aminobutyric acid (GABA)-B-R3 suppresses sugar reward.

(A) Cell type-specific GABA receptor silencing in the protocerebral anterior medial (PAM) neurons directed by R58E02-GAL4, showing GABA-B-R3-specific memory increment. *q < 0.008 (Benjamini and …

Figure 1—source data 1

GABA receptor silencing in the PAM neurons and the effects on appetitive memory.

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Figure 1—source data 2

Acute suppression of GABA-B-R3.

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Figure 1—source data 3

Memory acquisition curve in control and GABA-B-R3 knock-down flies.

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Figure 1—source data 4

Sugar response in the PAM neurons in control and GABA-B-R3 knock-down flies.

https://cdn.elifesciences.org/articles/64907/elife-64907-fig1-data4-v1.xlsx
Figure 1—figure supplement 1
Gamma-aminobutyric acid (GABA)-B-R3 knock-down in the protocerebral anterior medial (PAM) neurons increases memory strength.

(A) Appetitive memory score in R58E02-GAL4/UAS-GABA-B-R3-RNAi flies compared with parental controls. Mean ± SEM hereafter. *p<0.05 (Sidak’s test, N = 12). (B) Appetitive memory score in R58E02-GAL4/U…

Figure 1—figure supplement 1—source data 1

Appetitive memory score in control and GABA-B-R3 knock-down flies.

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Figure 1—figure supplement 1—source data 2

Appetitive memory score in control and GABA-B-R3 knock-down flies with a second RNAi line.

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Figure 1—figure supplement 1—source data 3

Memory acquisition curve in wild-type flies.

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Figure 2 with 1 supplement
Presynaptic GABAergic inhibition of the protocerebral anterior medial (PAM) neurons.

(A) A schematic showing brain regions visualized in subsequent figures. (B) A substack projection image of double labeling of GFP-tagged GABA-B-R3 protein and PAM neurons using GABA-B-R3-sfGFP-TVPTBF…

Figure 2—source data 1

Inhibition of GABA neurotransmission in APL neurons.

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Figure 2—source data 2

Sugar response in the axon terminals and dendrites of the PAM neurons.

https://cdn.elifesciences.org/articles/64907/elife-64907-fig2-data2-v1.xlsx
Figure 2—figure supplement 1
Presynaptic gamma-aminobutyric acid (GABA)-B-R3 localization and volumetric Ca2+ imaging from a defined protocerebral anterior medial (PAM) cell class.

(A) A substack projection image of double labeling of GFP-tagged GABA-B-R3 protein and PAM neurons using GABA-B-R3-sfGFP-TVPTBF, R58E02-GAL4/UAS-mCD8::RFP fly. Presynaptic localization of GABA-B-R3 …

Figure 3 with 1 supplement
Gamma-aminobutyric acid (GABA)-B-R3 controls presynaptic activity patterns in the protocerebral anterior medial (PAM) neurons.

(A) A single optical slice showing punctate sensor localization in the mushroom body (MB) lobe of R15A04-GAL4/UAS-Brp::GCaMP6s::mCherry flies. Scale bar, 10 μm. (B) The histograms of GFP/RFP signals …

Figure 3—source data 1

Temporal variance of spatially averaged intensities in control and GABA-B-R3 knock-down flies.

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Figure 3—source data 2

Probability distribution of active-zone calcium intensity before and during odor stimulation.

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Figure 3—source data 3

Average cluster size in control and GABA-B-R3 knock-down flies.

https://cdn.elifesciences.org/articles/64907/elife-64907-fig3-data3-v1.xlsx
Figure 3—figure supplement 1
Spatial and temporal characteristics of active-zone calcium signal in the presynapses of protocerebral anterior medial (PAM) neurons.

(A) Brp::GCaMP6s::mCherry (green) expressed by R15A04-GAL4 co-localizes with native Brp detected by nc82 (magenta). Scale bar, 1 μm. (B) Pseudo-color-coded distribution of active-zone calcium …

Figure 3—figure supplement 1—source data 1

Standard deviation of the active-zone calcium intensity.

https://cdn.elifesciences.org/articles/64907/elife-64907-fig3-figsupp1-data1-v1.xlsx
Figure 3—figure supplement 1—source data 2

obability distribution of active-zone calcium intensity before and during sugar stimulation.

https://cdn.elifesciences.org/articles/64907/elife-64907-fig3-figsupp1-data2-v1.xlsx
Figure 3—figure supplement 1—source data 3

Normalized peak size in control and GABA-B-R3 knock-down flies.

https://cdn.elifesciences.org/articles/64907/elife-64907-fig3-figsupp1-data3-v1.xlsx
Compromised odor specificity in the rewarded memory by attenuating local gamma-aminobutyric acid (GABA) signaling in the mushroom body.

(A–D) Olfactory generalization of appetitive memory depends on the blend ratio of a contaminant to a trained odor 4-methylcyclohexanol (4MCH) (A, C) or octan-3-ol (3OCT) (B, D). Significantly …

Figure 4—source data 1

Olfactory generalization of appetitive memory in control and GABA-B-R3 knock-down flies (4MCH).

https://cdn.elifesciences.org/articles/64907/elife-64907-fig4-data1-v1.xlsx
Figure 4—source data 2

Olfactory generalization of appetitive memory in control and GABA-B-R3 knock-down flies (3OCT).

https://cdn.elifesciences.org/articles/64907/elife-64907-fig4-data2-v1.xlsx
Figure 4—source data 3

Olfactory generalization of appetitive memory in control and Gad1 knock-down flies (4MCH).

https://cdn.elifesciences.org/articles/64907/elife-64907-fig4-data3-v1.xlsx
Figure 4—source data 4

Olfactory generalization of appetitive memory in control and Gad1 knock-down flies (3OCT).

https://cdn.elifesciences.org/articles/64907/elife-64907-fig4-data4-v1.xlsx
Author response image 1
Scatter plots showing dependency between the size (abscissa) and intensity (ordinate) of each peak calculated by the method we described.

Each circle represents a single cluster in the 9 frames before stimulation of a single representative fly.

Author response image 2
The cluster value map.

Each cluster is color-coded by the averaged intensity of associated pixels. Left: control, right: GABA-B-R3-KD.

Author response image 3
Time courses of normalized cluster size (left) and the comparison between genotypes using all frames from pre- and during-stimulation phases (frames 1-12, right).

The averaged cluster size was consistently higher in the GABA-B-R3 knock-down flies compared to wild-type flies. * P < 0.05, Mann Whitney test. N=15, 11.

Author response image 4
Normalized generalization curves using 4MCH:2MCH (left) and 3OCT:1OCT (right) mixtures.

Statistics suggest experimental group exhibit altered memory specificity to learned odors when compared with its parental control groups. (4MCH:2MCH) R58E02-GAL4/+, p = 0.030; UAS-GBR3-RNAi/+, p = …

Tables

Key resources table
Reagent type (species) or resourceDesignationSource or referenceIdentifiersAdditional information
Genetic reagent (Drosophila melanogaster)GMR58E02-GAL4Bloomington Drosophila Stock CenterBDSC: 41347; FLYB: FBal0253714; RRID:BDSC_41347
Genetic reagent (Drosophila melanogaster)GMR15A04-GAL4Bloomington Drosophila Stock CenterBDSC: 48671; FLYB: FBtp0057752; RRID:BDSC_48671
Genetic reagent (Drosophila melanogaster)VT043924-GAL4Vienna Drosophila Resource CenterVDRC: v201194; FLYB: FBtp0105273;
Genetic reagent (Drosophila melanogaster)UAS-GABA-B-R3-RNAiBloomington Drosophila Stock CenterBDSC: 50622; FLYB: FBti0157477; RRID:BDSC_50622FlyBase symbol:P{TRiP.HMC02989}attP40
Genetic reagent (Drosophila melanogaster)UAS-mCD8::GFPBloomington Drosophila Stock CenterBDSC: 32194; FLYB: FBti0131936; RRID:BDSC_32194FlyBase symbol: P{20XUAS-IVS-mCD8::GFP}
Genetic reagent (Drosophila melanogaster)UAS-GCaMP6sBloomington Drosophila Stock CenterBDSC: 42746; FLYB: FBti0151344; RRID:BDSC_42746FlyBase symbol: P{20XUAS-IVS-GCaMP6s}
Genetic reagent (Drosophila melanogaster)GABA-B-R3-sfGFP-TVPTBFBloomington Drosophila Stock CenterVDRC: v318614; FLYB: FBst0491638;FlyBase symbol: PBac{fTRG00613.sfGFP-TVPTBF}
Genetic reagent (Drosophila melanogaster)UAS-GCaMP6s.brpS.mCherryBloomington Drosophila Stock CenterBDSC: 77131; FLYB: FBtp0125966;
RRID:BDSC_77131
Antibody(Rabbit polyclonal) anti-GFPInvitrogenCat# A11122; RRID:AB_221569(1:1000)
Antibody(Mouse monoclonal) anti-THImmunoStarCat# 22941; RRID:AB_1267100(1:100)
Antibody(Rabbit polyclonal) anti-DsRedClontechCat# 632496; RRID:AB_10013483(1:200)
Antibody(Mouse monoclonal) anti-BrpDevelopmental Studies Hybridoma BankCat# nc82; RRID:AB_2314866(1:20)
Software, algorithmGraphPad Prism 5GraphPad Software, San Diego, CA, 2007https://www.graphpad.com/
Software, algorithmFijiMPI-CBGhttps://fiji.sc/

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