Sexual failure decreases sweet taste perception in male Drosophila via dopaminergic signaling
- Institute of Molecular Physiology, Shenzhen Bay Laboratory, China
- Department of Biology, Stanford University, United States
- Guangyang Bay Laboratory, Chongqing Institute for Brain and Intelligence, China
- School of Public Health, Capital Medical University, China
- Chinese Institutes for Medical Research, China
Figures
Figure 1 with 1 supplement
Sexual failure decreased sweet sensitivity.
(A) Schematic illustrating courtship conditioning strategy. (B) Percentage of males that performed feeding to 400 mM sucrose (n=40). (C) Fraction of Naïve, Failed, and Satisfied male flies showing proboscis extension reflex (PER) responses to different concentrations of sucrose. The PER experiment was performed immediately after conditioning unless otherwise indicated. Left: average PER responses of Naïve (blue), Failed (black), and Satisfied (red) flies (n=40–48). Right: MAT (mean acceptance threshold; the sucrose concentration where 50% of the flies show PER), plotted as a function of sexual state. (D) Fraction of male flies showing PER responses to different concentrations of sucrose. The PER experiment was performed immediately, 24 hr, 48 hr, and 72 hr after conditioning (n=46–135). Naïve and Satisfied showed pooled data from 0 to 72 hr post-treatment (E) Fraction of male flies showing PER responses to different concentrations of sucrose (n=40–48). Failed male flies were generated by two methods: one group was subjected to mated females (with cis-vaccenyl acetate, cVA), and the other was subjected to decapitated virgin females (without cVA). (F) Fraction of male flies showing PER responses to different concentrations of sucrose (n=55–60). Failed-re-copulated males were generated by placing individual failed males (within 30 min post-treatment) with 25 virgin females in a food vial for 4.5 hr. Data are shown as means ± SEM. Error bars represent SEM. n.s., p>0.05; *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001. One-way ANOVA (B–F MAT data) and two-way ANOVA (C–F PER data) were applied for statistical analysis.
-
Figure 1—source data 1
Source data for Figure 1.
- https://cdn.elifesciences.org/articles/105094/elife-105094-fig1-data1-v1.xlsx
Figure 1—figure supplement 1
Sexual failure decreased sweet sensitivity.
(A) Volume of 400 mM sucrose (left) and 800 mM (right) consumed by males fed ad libitum (n=31–58). (B) Starvation resistance of males was assayed in the presence of 5% sucrose (Fed) or 2% agar (Starved) (n=32–64). Yellow bars represent 12 hr light-on period. (C) Fraction of starved Naïve, Failed, and Satisfied male flies showing proboscis extension reflex (PER) responses to different concentrations of sucrose (n=40–60). (D) Fraction of Naïve, Failed, and Satisfied male flies showing PER responses to different concentrations of D-Fructose (n=48–60). (E) Fraction of male flies showing PER responses to different concentrations of D-Glucose (n=48–60). (F) Fraction of male flies showing PER responses to different concentrations of sucrose (n=40–70). Satisfied-Failed males were generated by placing individual Satisfied males (72 hr post-treatment) with 25 mated females in a food vial for 4.5 hr. Data are shown as means ± SEM. Error bars represent SEM. n.s., p>0.05; *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001. One-way ANOVA (A, C, F MAT data) and two-way ANOVA (B–F PER data) were applied for statistical analysis.
-
Figure 1—figure supplement 1—source data 1
Source data for Figure 1—figure supplement 1.
- https://cdn.elifesciences.org/articles/105094/elife-105094-fig1-figsupp1-data1-v1.xlsx
Figure 2 with 1 supplement
Dopamine signaling modulated the effect of sexual experience on sweet sensation.
(A) Proboscis extension reflex (PER) responses in male flies fed with 3IY, compared to male flies fed with vehicle (n=40–48). (B) PER responses in male flies with dopamine injection, compared to male flies injected with vehicle (n=40). (C, D) PER responses in male flies with indicated genotypes at 30°C (n=48–60). Data are shown as means ± SEM. Error bars represent SEM. n.s., p>0.05; *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. Two-way ANOVA (A–D PER data) and one-way ANOVA (A–D MAT data) were applied for statistical analysis.
-
Figure 2—source data 1
Source data for Figure 2.
- https://cdn.elifesciences.org/articles/105094/elife-105094-fig2-data1-v1.xlsx
Figure 2—figure supplement 1
Dopamine but not serotonin signaling modulated sexual experience-dependent sweet sensitivity.
(A) Proboscis extension reflex P(PER) responses in Th mutant male flies (n=40–42). (B) PER responses in male flies injected with various doses of dopamine (n=40). (C) PER responses in male flies fed with DL-p-chlorophenylalanine (pCPA) (n=48–60). (D) PER responses in male flies fed with methysergide (n=40–50). (E, F) PER responses in male flies with indicated genotypes at 20°C (n=40–80). Data are shown as means ± SEM. Error bars represent SEM. n.s., p>0.05; *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001. Two-way ANOVA (A–F PER data) and one-way ANOVA (A–F MAT data) were applied for statistical analysis.
-
Figure 2—figure supplement 1—source data 1
Source data for Figure 2—figure supplement 1.
- https://cdn.elifesciences.org/articles/105094/elife-105094-fig2-figsupp1-data1-v1.xlsx
Figure 3 with 1 supplement
Dopamine neurons modulated the activity of Gr5a+ neurons upon sexual failure.
(A) Schematic diagram of in vivo calcium imaging paradigm. (B) Quantification of the in vivo calcium responses of Gr5a+ neurons to the indicated concentrations of sucrose (n=5-18). (C) Averaged traces of the calcium responses to 200 mM sucrose. The horizontal black bar represents sucrose application. The solid lines represent the average trace, and shaded regions represent ± SEM. (D) Representative pseudocolor images of the calcium responses to 200 mM sucrose. The dashed area was quantified to measure the response. (E) Top: Representative images from the GRASP experiment between dopaminergic neurons and sweet Gustatory receptor neurons (GRNs). Bottom: Enlarged images of the subesophageal zone (SEZ) region seen on the top. Scale bars, 20 μm. (F) Average fluorescence reported by TH >nsyb-spGFP1-10, Gr5a>CD4-spGFP11 (normalized to Naive) in the SEZ. Regions of interest (ROIs) are circled by dashed lines in (E). Lines represent mean ± SEM, and dots indicate values for individual flies. (G) Top: Representative images of CaLexA-induced GFP reporter in dopaminergic neurons. Bottom: Enlarged images of the SEZ region were shown on top. Scale bars, 20 μm. (H, I) Average fluorescence reported by TH-GAL4/UAS-mLexA-NFAT, LexAop-CD2-GFP (normalized to Naive) in the SEZ (H), in the AL (I). Regions of interest (ROIs) are circled by dashed lines in (G). Lines represent mean ± SEM, and dots indicate values for individual flies. Data are shown as means ± SEM. Error bars represent SEM. n.s., p>0.05; *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001. Two-way ANOVA (B) and one-way ANOVA (F, H, I) were applied for statistical analysis.
-
Figure 3—source data 1
Source data for Figure 3.
- https://cdn.elifesciences.org/articles/105094/elife-105094-fig3-data1-v1.xlsx
Figure 3—figure supplement 1
Dopaminergic neurons had functional connections with sweet gustatory receptor neurons (GRNs).
(A) The expression of membrane-bound GFP (mCD8GFP, Green) in dopaminergic neurons driven by TH-Gal4 and membrane-bound RFP (rCD2RFP, Red) in Gr5a+ neurons driven by Gr5a-LexA. Scale bars, 20 μm. (B) Representative image of the GRASP experiment to show connections between dopaminergic neurons and sweet GRNs in TH >nsyb-spGFP1-10, Gr5a>CD4-spGFP11 flies (left). Genetic controls of GRASP experiments were shown on the center and right. Scale bars, 20 μm. (C, D) Averaged traces of the calcium responses to 200 mM sucrose in male flies with indicated genotypes are shown on the left. Quantification of the calcium responses was shown on the right (n=6–13). Top: Naïve. Bottom: Failed. The horizontal black bars represent sucrose application. The solid lines represent the average trace, and shaded regions represent ± SEM. One-way ANOVA (C, D) was applied for statistical analysis.
-
Figure 3—figure supplement 1—source data 1
Source data for Figure 3—figure supplement 1.
- https://cdn.elifesciences.org/articles/105094/elife-105094-fig3-figsupp1-data1-v1.xlsx
Figure 4 with 1 supplement
The effect of sexual failure was modulated through two dopamine receptors, Dop1R1 and Dop2R.
(A–E) Proboscis extension reflex (PER) responses in male flies with indicated genotypes (n=40–60). (F) Mean acceptance Threshold threshold (MAT) calculation for the PER data from (A–E), plotted as a function of sexual state and type of flies. Data are shown as means ± SEM. Error bars represent SEM. n.s., p>0.05; *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001. Two-way ANOVA (A–E) and one-way ANOVA (F) were applied for statistical analysis.
-
Figure 4—source data 1
Source data for Figure 4.
- https://cdn.elifesciences.org/articles/105094/elife-105094-fig4-data1-v1.xlsx
Figure 4—figure supplement 1
Dop1R1 and Dop2R were necessary for sexual failure-induced change in sweet sensitivity.
(A–D) Proboscis extension reflex (PER) responses in male flies with indicated genotypes (n=40–60). Data are shown as means ± SEM. Error bars represent SEM. n.s., p>0.05; *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. Two-way ANOVA (A–D PER data) and one-way ANOVA (A–D MAT data) were applied for statistical analysis.
-
Figure 4—figure supplement 1—source data 1
Source data for Figure 4—figure supplement 1.
- https://cdn.elifesciences.org/articles/105094/elife-105094-fig4-figsupp1-data1-v1.xlsx
Figure 5 with 1 supplement
Dop1R1+ and Dop2R+ neurons regulated sexual experience-dependent sweet sensitivity.
(A–D) Proboscis extension reflex (PER) responses in male flies with indicated genotypes (n=40). Data are shown as means ± SEM. Error bars represent SEM. n.s., p>0.05; *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. Two-way ANOVA (A–D PER data) and one-way ANOVA (A–D MAT data) were applied for statistical analysis.
-
Figure 5—source data 1
Source data for Figure 5.
- https://cdn.elifesciences.org/articles/105094/elife-105094-fig5-data1-v1.xlsx
Figure 5—figure supplement 1
Dop1R2+ or DopEcR+ neurons did not affect sweet sensitivity following sexual failure.
(A, B) Proboscis extension reflex (PER) responses in male flies with indicated genotypes (n=40). (C) Mean acceptance threshold (MAT), plotted as a function of sexual state and type of flies. Data are shown as means ± SEM. Error bars represent SEM. n.s., p>0.05; *p<0.05; **p<0.01; ***p<0.001, ****p<0.0001. Two-way ANOVA (A, B) and one-way ANOVA (C) were applied for statistical analysis.
-
Figure 5—figure supplement 1—source data 1
Source data for Figure 5—figure supplement 1.
- https://cdn.elifesciences.org/articles/105094/elife-105094-fig5-figsupp1-data1-v1.xlsx
Figure 6 with 1 supplement
Gr5a+ neurons received dopaminergic modulation through Dop1R1 and Dop2R.
(A, B) The expression of membrane-bound GFP (mCD8GFP, Green) in Gr5a+ neurons driven by Gr5a-LexA and membrane-bound RFP (mCD8RFP, Red) in dopamine receptor neurons driven by Dop1R1-Gal4 (A), and Dop2R-Gal4 (B). Scale bars, 20 μm. (C, D) PER responses in male flies with indicated genotypes (n=40). (E) Averaged traces of the calcium responses to 200 mM sucrose in male flies with indicated genotypes are shown on top. Quantification of the calcium responses was shown on the bottom (n=6–7). The horizontal black bars represent sucrose application. The solid lines represent the average trace, and shaded regions represent ± SEM. Data are shown as means ± SEM. Error bars represent SEM. n.s., p>0.05; *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. Two-way ANOVA (C, D PER data) and one-way ANOVA (C, D MAT data, E) were applied for statistical analysis.
-
Figure 6—source data 1
Source data for Figure 6.
- https://cdn.elifesciences.org/articles/105094/elife-105094-fig6-data1-v1.xlsx
Figure 6—figure supplement 1
Dopaminergic modulation of Gr5a+ neurons following sexual failure did not require DopEcR.
(A) The expression of membrane-bound GFP (mCD8GFP) in dopamine receptor neurons driven by Dop1R1-Gal4 (left), Dop2R-Gal4 (center), and DopEcR-Gal4 (right). Scale bars, 20 μm. (B) The expression of membrane-bound GFP (mCD8GFP, Green) in Gr5a+ neurons driven by Gr5a-LexA (left) and membrane-bound RFP (mCD8RFP, Red) in dopamine receptor neurons driven by DopEcR-Gal4. Scale bars, 20 μm. (C) Proboscis extension reflex (PER) responses in male flies with indicated genotypes (n=40). Data are shown as means ± SEM. Error bars represent SEM. n.s., p>0.05; *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001. Two-way ANOVA (C PER data) and one-way ANOVA (C MAT data) were applied for statistical analysis.
-
Figure 6—figure supplement 1—source data 1
Source data for Figure 6—figure supplement 1.
- https://cdn.elifesciences.org/articles/105094/elife-105094-fig6-figsupp1-data1-v1.xlsx
Figure 7
Working model for the sexual failure-dependent modulation of sweet perception.
In the subesophageal zone (SEZ) of the fly brain, dopaminergic neurons form synaptic connections with sweet-sensing Gr5a+ neurons, which express the dopamine receptors Dop1R1 and Dop2R. This connectivity enables Gr5a+ neurons to receive dopaminergic modulation, enhancing the flies' sweet sensitivity. However, after mating failure, the activity of these SEZ dopaminergic neurons is selectively inhibited, and their synaptic connections with Gr5a+ neurons are weakened. This reduction in dopaminergic signaling ultimately decreases the flies' sweet taste perception. This mechanism highlights a specific pathway by which unsuccessful mating experiences influence sensory processing through neural plasticity in dopamine-associated circuits.
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (D. melanogaster) | TH-GAL4 | PMID:30799021 | Gift from Yi Rao | |
| Genetic reagent (D. melanogaster) | UAS-dTRPA1 | other | Gift from David Anderson | |
| Genetic reagent (D. melanogaster) | UAS-Shits | other | Gift from David Anderson | |
| Genetic reagent (D. melanogaster) | Gr5a-GAL4 | Bloomington Drosophila Stock Center | Cat: #57592 | |
| Genetic reagent (D. melanogaster) | UAS-GCaMP6m | Bloomington Drosophila Stock Center | Cat: #42748 | |
| Genetic reagent (D. melanogaster) | Gr5a-LexA | Bloomington Drosophila Stock Center | Cat: #93014 | |
| Genetic reagent (D. melanogaster) | UAS-nSyb-spGFP1-10, lexAop-CD4-spGFP11 | Bloomington Drosophila Stock Center | Cat: #64314 | |
| Genetic reagent (D. melanogaster) | UAS-CalexA | Bloomington Drosophila Stock Center | Cat: #66542 | |
| Genetic reagent (D. melanogaster) | Dop1R1-/- mutant | PMID:30799021 | Gift from Yi Rao | |
| Genetic reagent (D. melanogaster) | Dop2R-/- mutant | PMID:30799021 | Gift from Yi Rao | |
| Genetic reagent (D. melanogaster) | Dop1R2-/- mutant | PMID:30799021 | Gift from Yi Rao | |
| Genetic reagent (D. melanogaster) | DopEcR-/- mutant | PMID:30799021 | Gift from Yi Rao | |
| Genetic reagent (D. melanogaster) | Dop1R1-GAL4 | PMID:30799021 | Gift from Yi Rao | |
| Genetic reagent (D. melanogaster) | Dop2R-GAL4 | PMID:30799021 | Gift from Yi Rao | |
| Genetic reagent (D. melanogaster) | UAS-NaChBac | Bloomington Drosophila Stock Center | Cat: #9469 | |
| Genetic reagent (D. melanogaster) | UAS-Kir2.1 | Bloomington Drosophila Stock Center | Cat: #6595 | |
| Genetic reagent (D. melanogaster) | 10×UAS-IVS-mCD8::RFP, 13×LexAop2-mCD8::GFP | Bloomington Drosophila Stock Center | Cat: #32229 | |
| Genetic reagent (D. melanogaster) | UAS-GCaMP6m | Bloomington Drosophila Stock Center | Cat: #42750 | |
| Genetic reagent (D. melanogaster) | UAS-Dop1R1 RNAi | Tsinghua Fly Center | Cat: #4278 | |
| Genetic reagent (D. melanogaster) | UAS-Dop2R RNAi | Tsinghua Fly Center | Cat: #2141 | |
| Genetic reagent (D. melanogaster) | Th-/- mutant | PMID:30799021 | Gift from Yi Rao | |
| Genetic reagent (D. melanogaster) | LexAop-rCD2RFP, UAS-mCD8GFP | Bloomington Drosophila Stock Center | Cat: #67093 | |
| Genetic reagent (D. melanogaster) | LexAop-GCaMP6m | other | Gift from Yufeng Pan | |
| Genetic reagent (D. melanogaster) | UAS-CsChrimson | other | Gift from Yufeng Pan | |
| Genetic reagent (D. melanogaster) | elav-GAL4 | Bloomington Drosophila Stock Center | Cat: #25750 | |
| Genetic reagent (D. melanogaster) | UAS-Dop1R2 RNAi | Tsinghua Fly Center | Cat: #5285 | |
| Genetic reagent (D. melanogaster) | UAS-DopEcR RNAi | Tsinghua Fly Center | Cat: #3275 | |
| Genetic reagent (D. melanogaster) | Dop1R2-GAL4 | PMID:30799021 | Gift from Yi Rao | |
| Genetic reagent (D. melanogaster) | DopEcR-GAL4 | PMID:30799021 | Gift from Yi Rao | |
| Genetic reagent (D. melanogaster) | UAS-mCD8::GFP | Bloomington Drosophila Stock Center | Cat: #5137 | |
| Antibody | anti-GFP (mouse) | Sigma-Aldrich | Cat: #G6539 | |
| Antibody | anti-GFP (mouse) | Abcam | Cat: #ab1218 | |
| Antibody | anti-DsRed (rabbit) | Takara Bio | Cat: #632496 | |
| Antibody | anti-GFP (rabbit) | Thermo Fisher Scientific | Cat: #A11122 | |
| Antibody | Goat anti-mouse Alexa Fluor 488 | Thermo Fisher Scientific | Cat: #A11029 | |
| Antibody | Goat anti-rabbit Alexa Fluor 546 | Thermo Fisher Scientific | Cat: #A11010 | |
| Antibody | Goat anti-rabbit Alexa Fluor 488 | Thermo Fisher Scientific | Cat: #A11034 | |
| Chemical compound | Sucrose | Sigma-Aldrich | Cat: #S0389 | |
| Chemical compound | 3-Iodo-L-tyrosine | Sigma-Aldrich | Cat: #I8250 | |
| Chemical compound | Dopamine | Aladdin | Cat: #A303863 | |
| Chemical compound | NaCl | Sigma-Aldrich | Cat: #S3014 | |
| Chemical compound | MgCl2 | Sigma-Aldrich | Cat: #M4880 | |
| Chemical compound | NaHCO3 | Sigma-Aldrich | Cat: #S5761 | |
| Chemical compound | NaH2PO4 | Sigma-Aldrich | Cat: #S3139 | |
| Chemical compound | KCl | Sigma-Aldrich | Cat: #P9541 | |
| Chemical compound | CaCl2 | Sigma-Aldrich | Cat: #C5670 | |
| Chemical compound | HEPES | Sigma-Aldrich | Cat: #54457 | |
| Chemical compound | Paraformaldehyde | Electron Microscopy | Cat: #15713 | |
| Chemical compound | Calf serum | Thermo Fisher Scientific | Cat: #16010159 | |
| Chemical compound | Triton X-100 | Sigma-Aldrich | Cat: #T8787 | |
| Chemical compound | D-Fructose | Tokyo Chemical Industry | Cat: #F0060 | |
| Chemical compound | D-Glucose | Tokyo Chemical Industry | Cat: #G0048 | |
| Chemical compound | DL-p-chlorophenylalanine | Sigma-Aldrich | Cat: #c6506 | |
| Chemical compound | Methysergide maleate | Abcam | Cat: #ab120530 | |
| Chemical compound | All trans-retinal | Sigma-Aldrich | Cat: #R2500 | |
| Commercial assay or kit | Graduated glass capillary | VWR | Cat: #53432–604 | |
| Software, algorithm | Fiji/ImageJ | NIH | RRID:SCR_003070 | |
| Software, algorithm | GraphPad Prism 9 | GraphPad Software | RRID:SCR_002798 |
Additional files
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Sexual failure decreases sweet taste perception in male Drosophila via dopaminergic signaling
eLife 14:RP105094.
https://doi.org/10.7554/eLife.105094.3
