(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 …
GABA receptor silencing in the PAM neurons and the effects on appetitive memory.
Acute suppression of GABA-B-R3.
Memory acquisition curve in control and GABA-B-R3 knock-down flies.
Sugar response in the PAM neurons in control and GABA-B-R3 knock-down flies.
(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…
Appetitive memory score in control and GABA-B-R3 knock-down flies.
Appetitive memory score in control and GABA-B-R3 knock-down flies with a second RNAi line.
Memory acquisition curve in wild-type flies.
(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…
Inhibition of GABA neurotransmission in APL neurons.
Sugar response in the axon terminals and dendrites of the PAM neurons.
(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 …
(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 …
Temporal variance of spatially averaged intensities in control and GABA-B-R3 knock-down flies.
Probability distribution of active-zone calcium intensity before and during odor stimulation.
Average cluster size in control and GABA-B-R3 knock-down flies.
(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 …
Standard deviation of the active-zone calcium intensity.
obability distribution of active-zone calcium intensity before and during sugar stimulation.
Normalized peak size in control and GABA-B-R3 knock-down flies.
(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 …
Olfactory generalization of appetitive memory in control and GABA-B-R3 knock-down flies (4MCH).
Olfactory generalization of appetitive memory in control and GABA-B-R3 knock-down flies (3OCT).
Olfactory generalization of appetitive memory in control and Gad1 knock-down flies (4MCH).
Olfactory generalization of appetitive memory in control and Gad1 knock-down flies (3OCT).
Each circle represents a single cluster in the 9 frames before stimulation of a single representative fly.
Each cluster is color-coded by the averaged intensity of associated pixels. Left: control, right: GABA-B-R3-KD.
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.
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 = …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Genetic reagent (Drosophila melanogaster) | GMR58E02-GAL4 | Bloomington Drosophila Stock Center | BDSC: 41347; FLYB: FBal0253714; RRID:BDSC_41347 | |
Genetic reagent (Drosophila melanogaster) | GMR15A04-GAL4 | Bloomington Drosophila Stock Center | BDSC: 48671; FLYB: FBtp0057752; RRID:BDSC_48671 | |
Genetic reagent (Drosophila melanogaster) | VT043924-GAL4 | Vienna Drosophila Resource Center | VDRC: v201194; FLYB: FBtp0105273; | |
Genetic reagent (Drosophila melanogaster) | UAS-GABA-B-R3-RNAi | Bloomington Drosophila Stock Center | BDSC: 50622; FLYB: FBti0157477; RRID:BDSC_50622 | FlyBase symbol:P{TRiP.HMC02989}attP40 |
Genetic reagent (Drosophila melanogaster) | UAS-mCD8::GFP | Bloomington Drosophila Stock Center | BDSC: 32194; FLYB: FBti0131936; RRID:BDSC_32194 | FlyBase symbol: P{20XUAS-IVS-mCD8::GFP} |
Genetic reagent (Drosophila melanogaster) | UAS-GCaMP6s | Bloomington Drosophila Stock Center | BDSC: 42746; FLYB: FBti0151344; RRID:BDSC_42746 | FlyBase symbol: P{20XUAS-IVS-GCaMP6s} |
Genetic reagent (Drosophila melanogaster) | GABA-B-R3-sfGFP-TVPTBF | Bloomington Drosophila Stock Center | VDRC: v318614; FLYB: FBst0491638; | FlyBase symbol: PBac{fTRG00613.sfGFP-TVPTBF} |
Genetic reagent (Drosophila melanogaster) | UAS-GCaMP6s.brpS.mCherry | Bloomington Drosophila Stock Center | BDSC: 77131; FLYB: FBtp0125966; RRID:BDSC_77131 | |
Antibody | (Rabbit polyclonal) anti-GFP | Invitrogen | Cat# A11122; RRID:AB_221569 | (1:1000) |
Antibody | (Mouse monoclonal) anti-TH | ImmunoStar | Cat# 22941; RRID:AB_1267100 | (1:100) |
Antibody | (Rabbit polyclonal) anti-DsRed | Clontech | Cat# 632496; RRID:AB_10013483 | (1:200) |
Antibody | (Mouse monoclonal) anti-Brp | Developmental Studies Hybridoma Bank | Cat# nc82; RRID:AB_2314866 | (1:20) |
Software, algorithm | GraphPad Prism 5 | GraphPad Software, San Diego, CA, 2007 | https://www.graphpad.com/ | |
Software, algorithm | Fiji | MPI-CBG | https://fiji.sc/ |
List of fly strains and crosses for experiments, and the statistical results.