A neuropeptide regulates fighting behavior in Drosophila melanogaster
- State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, China
- University of Chinese Academy of Sciences, China
- Chinese Institute for Brain Research, Peking-Tsinghua Center for Life Sciences, Zhongguangchun Life Sciences Park, China
- Shenzhen Bay Laboratory, China
- State Key Laboratory of Membrane Biology, College of Life Sciences, IDG/McGovern Institute for Brain Research, Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Center for Quantitative Biology, Academy for Advanced Interdisciplinary Studies, Peking University, China
- School of Life Sciences, University of Science and Technology of China, China
- Peking-Tsinghua Center for Life Sciences, PKU-IDG/McGovern Institute for Brain Research, Advanced Innovation Center for Genomics, Peking University School of Life Sciences, China
Figures
Figure 1 with 6 supplements
The Dsk Gene Is Essential for Modulating Male-Male Aggression.
(A) Generation of ΔDsk (A1) and ΔDskGAL4 (A2). Dashed boxes indicate the region replaced by 3P3-RFP (A1) or GAL4 (A2) cassette. (B) Male adult brains of the indicated genotypes were stained with …
Figure 1—figure supplement 1
Molecular validation of mutants for Dsk.
(A) Quantification of Dsk mRNA by qRT-PCR in the homozygotes, heterozygotes, and wildtypes. The Dsk transcripts were not detectable in ΔDsk /ΔDsk and ΔDskGAL4/ΔDskGAL4 mutants. Triplicate assays per …
Figure 1—figure supplement 2
ΔDsk suppresses wing threat frequency in males.
(A) ΔDsk mutants show reduced number of wing threats compared to wildtype and heterozygous controls. ***p<0.001 (Kruskal-Wallis and post-hoc Mann-Whitney U tests).
Figure 1—figure supplement 3
Aggressive behavior was promoted by Dsk overexpression.
(A–B) RU486 induced overexpression of Dsk increased number of lunges per minute after initiation (A) and shortened fighting latency (B) in males. **p<0.01, ***p<0.001, n.s. indicates no significant …
Figure 1—figure supplement 4
DSK signal modulates female-female aggression.
(A) Head butts frequency of female-female aggression. Homozygotes of ΔDsk showed a significant decrease in head butts compared to heterozygotes or wildtypes. (B) Fighting latency of female-female …
Figure 1—figure supplement 5
Courtship, locomotion and feeding behavior of ΔDsk mutants.
(A) Courtship indices for male-male courtship. (B) Courtship indices for male-female courtship. (C) The locomotive speed of ΔDsk mutants. (D) The feeding behavior of ΔDsk mutants. ΔDsk mutants show …
Figure 1—figure supplement 6
The design of aggression chamber.
The aggression chamber consists of four acrylic plates. The bottom plate (Plate 4) contains 12 wells for food patches (diameter: 8 mm; depth: 3 mm). The lower (Plate 3) and upper plates (Plate 2) …
Figure 2 with 4 supplements
DskGAL4 Neurons Modulate Male-Male Aggressive Behavior.
(A) Generation of DskGAL4 Knock-in line. GAL4 was fused to the end of open reading frame (ORF) of Dsk with a T2A peptide linker. (B–C) Anatomical features of DSK neurons revealed by DskGAL4 driven UA…
Figure 2—figure supplement 1
Projection patterns of DskGAL4 neurons in female.
(A–D) Characterization of individual DskGAL4 neurons using MultiColor FlpOut (MCFO) method in females (A). Eight DSK-expressing neurons were classified into three neuronal cell types (B–D). Scale …
Figure 2—figure supplement 2
Locomotion behavior of TNT and trpA1 experiments.
(A) The locomotive speed of TNT experiment. (B) The locomotive speed of trpA1 experiment. Inactivation or activation of DskGAL4 neurons could not affect locomotion behavior. ns. indicates no …
Figure 2—figure supplement 3
The type I and type II neurons are necessary for aggression.
(A) A negative correlation between the number of type I plus type II neurons inhibited and the lunge numbers of individual flies. (B) A negative correlation between the number of type I neurons …
Figure 2—figure supplement 4
The design of aggression chamber.
The aggression chamber consists of four acrylic plates. The bottom plate (Plate 4) contains 12 wells for food patches (diameter: 8 mm; depth: 3 mm). The lower (Plate 3) and upper plates (Plate 2) …
Figure 3 with 5 supplements
Dsk Receptor CCKLR-17D1 Is Necessary for Male-Male Aggression.
(A) Schematic view of CCKLR-17D1 gene locus and generation of ΔCCKLR-17D1 and CCKLR-17D1GAL4 Knock-in lines. The last two exons of CCKLR-17D1 gene were replaced by 3P3-RFP to generate ΔCCKLR-17D1. …
Figure 3—figure supplement 1
Molecular validation of mutants for Dsk receptors.
(A) Quantification of CCKLR-17D1 and CCKLR-17D3 mRNA by qRT-PCR in the CCKLR mutants and wildtypes. CCKLR-17D1 and CCKLR-17D3 transcripts were not detected in ΔCCKLR-17D1/y and ΔCCKLR-17D3/y …
Figure 3—figure supplement 2
Expression patterns of CCKLR-17D3GAL4.
(A) Generation of CCKLR-17D3GAL4 Knock-in line. GAL4 was fused to the end of open reading frame (ORF) of CCKLR-17D3 with a T2A peptide linker. (B–C) Projection patterns of CCKLR-17D3GAL4 neurons in …
Figure 3—figure supplement 3
ΔCCKLR-17D1 does not affect locomotion behavior.
(A) The locomotive speed of ΔCCKLR-17D1 and ΔCCKLR-17D3 mutants. ns. indicates no significant difference (Kruskal-Wallis and post-hoc Mann-Whitney U tests).
Figure 3—figure supplement 4
Aggression, locomotion behavior of CCKLR-17D1 RNAi in male flies.
(A–B) RNAi knock-down of CCKLR-17D1 decreases inter-male aggression. (A) Number of lunges per minute after initiation. (B) Fighting latency. elav-GAL4; UAS-dcr2 or CCKLR-17D1GAL4 drives CCKLR-17D1 …
Figure 3—figure supplement 5
The design of aggression chamber.
The aggression chamber consists of four acrylic plates. The bottom plate (Plate 4) contains 12 wells for food patches (diameter: 8 mm; depth: 3 mm). The lower (Plate 3) and upper plates (Plate 2) …
Figure 4 with 3 supplements
DSK Functions Downstream of a subset of P1 Neurons to Modulate Fighting Behavior.
(A) Transsynaptic labeling by trans-Tango method identifies DSK neurons as postsynaptic partners of a subset of P1 neurons. Expression of the trans-Tango ligand in a subset of P1 neurons (red) (A1) …
Figure 4—figure supplement 1
Controls for the trans-Tango and GRASP experiments.
(A) Negative control demonstrates lack of postsynaptic signals (anti-HA, magenta) (A2) for the trans-Tango experiment. Red, anti-DSK antibody staining (A1). (B–C) Negative controls for the GRASP …
Figure 4—figure supplement 2
Activation of a subset of P1 neurons could promote courtship in the ΔDsk mutant background.
(A–B) Raster plots of unilateral wing extensions during 10 min. (C–D) The number (C) and total duration (D) of unilateral wing extensions in (A–B) during thermogenetic activation of a subset of P1 …
Figure 4—figure supplement 3
The design of aggression chamber.
The aggression chamber consists of four acrylic plates. The bottom plate (Plate 4) contains 12 wells for food patches (diameter: 8 mm; depth: 3 mm). The lower (Plate 3) and upper plates (Plate 2) …
Figure 5 with 1 supplement
The Functional Connectivity Between DSK Neurons and R71G01-LexA labeled P1 Neurons.
(A) Illustration of patch-clamp recording on DSK neurons (magenta). ATP-gated ion channel P2X2 was expressed in R71G01-LexA labeled P1 neurons for chemogenetic activation of P1 neurons. Genotype: +/y…
Figure 5—figure supplement 1
R71G01-LexA labeled P1 neurons do not respond to activation of DSK neurons.
(A–B) Calcium imaging of GCaMP6m in the cell bodies of R71G01-LexA labeled P1 neurons with P2X2-expressing DSK neurons activated by ATP. ATP: 2.5 mM. White dashed lines indicate ROI. Scale bars …
Figure 6 with 1 supplement
Conditional overexpression of DSK promotes winner effect and Winner Show Increased Calcium Activity of DSK-M Neurons.
(A–C) Conditionally overexpression of Dsk increases number of lunges compared to controls. Pairs of flies of the indicated genotypes were introduced into one chamber, one of which overexpressed DSK …
Figure 6—figure supplement 1
The design of aggression chamber.
The aggression chamber consists of four acrylic plates. The bottom plate (Plate 4) contains 12 wells for food patches (diameter: 8 mm; depth: 3 mm). The lower (Plate 3) and upper plates (Plate 2) …
Videos
Video 1
Left, female aggression of ΔDsk mutants.
Right, female aggression of wildtypes. Female ΔDsk mutants show fewer aggressive encounters than wildtypes.
Video 2
Thermogenetic activation of DskGAL4 neurons with UAS-dTRPA1 promotes inter-male aggression.
Video 3
Chemogenetic activation of P1 neurons by ATP elicits calcium responses from DskGAL4 neurons.
Additional files
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Supplementary file 1
Key resources table.
- https://cdn.elifesciences.org/articles/54229/elife-54229-supp1-v2.docx
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Supplementary file 2
The number of labeled cells of mosaic flies.
- https://cdn.elifesciences.org/articles/54229/elife-54229-supp2-v2.xls
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Transparent reporting form
- https://cdn.elifesciences.org/articles/54229/elife-54229-transrepform-v2.docx
