Selective lifelong suppression of an odor processing channel in response to critical period experience
- Department of Neurosciences, Case Western Reserve University School of Medicine, United States
- Department of Biology, Case Western Reserve University, United States
Figures
Figure 1
Lifelong suppression of Or42a OSNs after critical-period EB exposure.
(A) Schematic of the Drosophila olfactory circuit at the level of the AL. OSN, olfactory sensory neuron, PN, projection neuron, LN, local interneuron, EG, ensheathing glia. (B) Overview of odorant-exposure experiments. Flies are exposed to 15% (v/v) ethyl butyrate (EB) or mineral oil vehicle control during the Or42a OSN critical period from 0 to 2 days post-eclosion (DPE). (C–F) Live imaging of Or42a OSN odor-evoked activity at 2 (C, D) or 25 (E, F) DPE, following exposure to 15% EB or mineral oil from 0 to 2 DPE. Or42a-GAL4>UAS-GCaMP8f flies were stimulated with three 1 s puffs (yellow bars) of 5% EB (red traces and data points) or mineral oil vehicle control (gray traces and data points) spaced 10 s apart. Traces (C, E) show the mean ΔF/F0 values of all trials. Data points (D, F) represent the mean peak ΔF/F0 values for each fly. (G–H) Live imaging of ASAP5 responses in Or42a OSNs in 2 DPE flies following exposure to 15% EB or mineral oil from 0 to 2 DPE. Flies were stimulated with odor puffs as above. Traces (G) represent the mean ΔF/F0 values of all trials. Data points (H) represent the mean peak ΔF/F0 values for each animal. Data in (D, F), and (H) are mean± SD. *p<0.05, **p<0.01, unpaired t-test (D) or Mann–Whitney U-test (F, H). Genotypes, raw values, and detailed statistics are provided in Figure 1—source data 1.
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Figure 1—source data 1
Numerical data used to generate Figure 1D and F.
- https://cdn.elifesciences.org/articles/108236/elife-108236-fig1-data1-v1.xlsx
Figure 2
Experience-dependent pruning in VM7 is restricted to the presynaptic compartment.
(A–B) Live imaging of VM7 projection neuron (PN) odor-evoked activity at 2 days post-eclosion (DPE), following exposure to 15% ethyl butyrate (EB) or mineral oil from 0 to 2 DPE. VM7 PN split-GAL4 >UAS-GCaMP8f flies were stimulated with three 1 s puffs (yellow bars) of 5% EB (red traces and data points) or mineral oil vehicle control (gray traces and data points) spaced 10 s apart. Traces (A) show the mean ΔF/F0 values of all trials. Data points (B) represent the mean peak ΔF/F0 values for each fly. (C) Representative dendritic arbors of VM7 PNs endogenously expressing the V5-tagged postsynaptic marker Discs large 1 (Dlg1), from flies exposed to mineral oil or 15% EB from 0 to 2 DPE. (D) Normalized counts of Dlg1 puncta. (E) Representative images of VM7 PNs after exposure to mineral oil or 15% EB from 0 to 2 DPE. Neuropil is visualized with antibody staining for the presynaptic active zone protein Bruchpilot (Brp). PN dendritic arbors occupying VM7 are enclosed in white boxes. (F) Volumetric measurements of VM7 PN dendritic arbors shown in (E). (G) Representative images of VM7 PN axonal arbors after exposure to mineral oil or 15% EB from 0 to 2 DPE. (H) Volumetric measurements of VM7 PN axonal arbors shown in (G). (I) Models of VM7 PN presynapses from (G) after exposure to mineral oil or 15% EB from 0 to 2 DPE, defined as Brp puncta masked by mCD8::GFP-labeled VM7 PNs. (J) Normalized counts of VM7 PN presynapses from (I). Data are mean± SD. ns, p≥0.05, *p<0.05, Mann–Whitney U-test. Genotypes, raw values, and detailed statistics are provided in Figure 2—source data 1.
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Figure 2—source data 1
Numerical data used to generate Figure 2B, D, F, H and J.
- https://cdn.elifesciences.org/articles/108236/elife-108236-fig2-data1-v1.xlsx
Figure 3
Experience-dependent pruning and inhibition are decoupled in Or43b olfactory sensory neurons (OSNs).
(A) DoOR 2.0 profile of ethyl butyrate (EB) responses across all Drosophila odorant receptors (ORs). (B–C) Representative images (bottom) and quantification of presynaptic content (B) and volume (C) of VM2 (Or43b OSNs) and VM7 (Or42a OSNs) in flies exposed to mineral oil or 15% EB from 0 to 2 days post-eclosion (DPE). Presynapses were visualized with Bruchpilot (Brp) staining. Data are mean± SD. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, unpaired t-test. (D–E) Two-photon imaging of GCaMP8f responses in Or43b OSNs at 2 DPE following exposure to 15% EB or mineral oil from 0 to 2 DPE. Awake flies were stimulated with three 1 s odor puffs (yellow bars) of 5% EB (red traces and data points) or mineral oil (gray traces and data points), spaced 10 s apart. Traces (D) represent the mean ΔF/F0 values of all trials. Data points (E) represent the mean peak ΔF/F0 values for each animal. Genotypes, raw values, and detailed statistics are provided in Figure 3—source data 1.
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Figure 3—source data 1
Numerical data used to generate Figure 3A, B, C and E.
- https://cdn.elifesciences.org/articles/108236/elife-108236-fig3-data1-v1.xlsx
Figure 4
Differential patterns of local interneuron (LN) innervation may contribute to interglomerular differences in experience-dependent glial pruning.
(A) Numbers of neurons within the cell types included in (B) and (C). (B–C) Sankey diagrams of the synaptic inputs and outputs of Or42a (B) and Or43b (C) olfactory sensory neurons (OSNs). Inputs were thresholded at 99% of all synapses and outputs were thresholded at 95% of all synapses. Brackets contain the number of synapses and percentage of total synapses for that category. All connectomic analyses are derived from the FlyWire FAFB dataset. Synapse counts are provided in Figure 4—source data 1.
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Figure 4—source data 1
Numerical data used to generate Figure 4B and C.
- https://cdn.elifesciences.org/articles/108236/elife-108236-fig4-data1-v1.xlsx
Figure 5
GABAergic inputs restrain glial pruning.
(A–B) Representative images (bottom) and quantification of VM7 (A) and VM2 (B) volume in flies exposed to mineral oil or 15% ethyl butyrate (EB) from 0 to 2 days post-eclosion (DPE). Data are mean ± SD. *p<0.05, **p<0.01, ****p<0.0001, one-way ANOVA. Genotypes, raw values, and detailed statistics are provided in Figure 5—source data 1.
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Figure 5—source data 1
Numerical data used to generate Figure 5A and B.
- https://cdn.elifesciences.org/articles/108236/elife-108236-fig5-data1-v1.xlsx
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (D. melanogaster) | UAS-GCaMP8f | BDSC | BDSC Stock #92587; RRID:BDSC_92587; FlyBase ID: FBst0092587 | Genotype: w[1118]; PBac{y[+mDint2] w[+mC]=20XUAS-IVS-jGCaMP8f}VK00005 |
| Genetic reagent (D. melanogaster) | UAS-ASAP5 | BDSC | BDSC Stock #605337; RRID:BDSC_605337; FlyBase ID: FBst0605337 | Genotype: w[*]; S (1)/CyO; PBac{y[+mDint2] w[+mC]=20XUAS-ASAP5}VK00005/TM6B, Tb(1) |
| Genetic reagent (D. melanogaster) | Or42a-mCD8::GFP | Golovin et al., 2019 | Broadie lab | Genotype: w; Or42a-mCD8::GFP |
| Genetic reagent (D. melanogaster) | Or43b-GAL4 | BDSC | BDSC Stock #23894; RRID:BDSC_23894; FlyBase ID: FBst0023894 | Genotype: w[*]; P{w[+mC]=Or43b-GAL4.C}110t8.1 |
| Genetic reagent (D. melanogaster) | Or42a-GAL4 | BDSC | BDSC Stock #9970; RRID:BDSC_9970; FlyBase ID: FBst0009970 | Genotype: w[*]; P{w[+mC]=Or42 a-GAL4.F}48.3B |
| Genetic reagent (D. melanogaster) | dlg1[4 K] | Parisi et al., 2023 | Mosca lab | Genotype: dlg1[4 K], UAS-Flp[8208] |
| Genetic reagent (D. melanogaster) | UAS-mCD8::GFP | BDSC | BDSC Stock #32186; RRID:BDSC_32186; FlyBase ID: FBst0032186 | Genotype: w*; P{10XUAS-IVS-mCD8::GFP}attP40 |
| Genetic reagent (D. melanogaster) | VM7 PN split-GAL4 | Xie et al., 2021 | Janelia FlyLight Split-GAL4 Driver Collection, line SS01265 | Genotype: w; VT033006-p65ADZp in attP40; R19G08-ZpGdbd in attP2/TM6B |
| Genetic reagent (D. melanogaster) | UAS-LexA RNAi | BDSC | BDSC Stock # 67947; RRID:BDSC_67947; FlyBase ID: FBst0067947 | Genotype: y(1) sc[*] v(1) sev[21]; P{y[+t7.7] v[+t1.8]=TRiP.HMS05772}attP40 |
| Genetic reagent (D. melanogaster) | UAS-rdl RNAi | BDSC | BDSC Stock # 31286; BDSC_31286; FlyBase ID: FBst0031286 | Genotype: y(1) v(1); P{y[+t7.7] v[+t1.8]=TRiP.JF01227}attP2 |
| Antibody | Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 647 | Thermo Fisher Scientific | Cat #A32728; RRID:AB_2866490 | Used 1:400 (IHC) |
| Antibody | anti-Bruchpilot (Mouse monoclonal) | Developmental Studies Hybridoma Bank | Cat #nc82; RRID:AB_2314866 | Used 1:50 (IHC) |
| Antibody | anti-GFP, Alexa Fluor 488 (Rabbit polyclonal) | Thermo Fisher Scientific | Cat #A21311; RRID:AB_221477 | Used 1:200 (IHC) |
| Antibody | anti-V5 tag, Alexa Fluor 488 (Mouse monoclonal) | Thermo Fisher Scientific | Cat #37–7500 A488; RRID:AB_2610630 | Used 1:100 (IHC) |
| Software, algorithm | Imaris | Bitplane | RRID:SCR_007370 | |
| Software, algorithm | FlyWire | https://flywire.ai/ | RRID:SCR_019205 | |
| Software, algorithm | fafbseg | https://github.com/navis-org/fafbseg-py; Schlegel, 2026; Dorkenwald et al., 2024; Schlegel et al., 2024 | RRID:SCR_008394 | Python package |
| Software, algorithm | DoOR.data | https://github.com/ropensci/DoOR.data; Münch, 2026a | RRID:SCR_000432 | R package for DoOR data |
| Software, algorithm | DoOR.functions | https://github.com/ropensci/DoOR.functions; Münch, 2026b | RRID:SCR_000432 | R package for DoOR functions |
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Selective lifelong suppression of an odor processing channel in response to critical period experience
eLife 14:RP108236.
https://doi.org/10.7554/eLife.108236.3
