Drosophila seminal sex peptide associates with rival as well as own sperm, providing SP function in polyandrous females
- Department of Molecular Biology and Genetics, Cornell University, United States
Figures
Figure 1
SP from a second male can bind to SP-deficient sperm of previous male stored within a mated female.
Cartoon: Pictorial representation of the crossing scheme (fly images from Biorender). Wild type (CS) females were first mated to an SP-null male and then, at the indicated time, to a spermless (SOT) …
Figure 2 with 1 supplement
Sperm from a second male are not bound to SP from a prior spermless male.
(Cartoon): Pictorial representation of the cross (fly images from Biorender); it is reciprocal of that in Figure 1. Females mated first with spermless (SOT) males and then a day later with SP-null …
Figure 2—figure supplement 1
Cartoon: Pictorial representation of cross (fly images from Biorender).
Females mated first with spermless (sot) male and then 3–6 hr ASFM with SP-null male that provided sperm. Panel: Sperm from SR of females mated to spermless males and then remated to SP-null males, …
Figure 3 with 1 supplement
Remating with spermless males restores fertility, delays receptivity and optimizes efficient sperm release in females that previously mated to SP-null males.
(A) Graphical representation of numbers of progeny produced by each female over the span of 10 days, following mating to control (TM3 siblings of SP-null males: SP+; red), SP-null males (SP-null; …
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Figure 3—source data 1
Source data for Figure 3D, E.
- https://cdn.elifesciences.org/articles/58322/elife-58322-fig3-data1-v2.xlsx
Figure 3—figure supplement 1
Western blot probed for SP.
Lanes/samples are 1: Fv, reproductive tract (RT) of three virgin females (negative control); 2: M, one pair of male accessory glands (positive control); 3: SP+, RT of three females mated to control …
Figure 4 with 1 supplement
SP from a male who also provides sperm can bind to SP-deficient sperm as well as to the donor’s sperm.
Cartoon (I): Pictorial representation of the experimental cross (fly images from Biorender). Females mated to SP-null males were remated to control (ProtB-dsRed) males at 1d ASFM. (A) Sperm from …
Figure 4—figure supplement 1
Little to no SP is transferred by multiply-mated males.
(A) Western blot probed for SP. Lanes/samples are 1: Fv, reproductive tract (RT) of two virgin females (negative control); 2: M, one pair of male accessory glands from a 3-day-old unmated virgin …
Figure 5
Sperm do not bind detectable LTR-SFPs from a second male.
Females mated to wild type (CS) males at 2 hr ASM show LTR-SFPs bound to sperm, CG1656 (A), CG1652 (E), CG9997 (I). Females mated to SP-null males show the same (B,F,J) but by 1d postmating …
Figure 6 with 1 supplement
Sperm received from SP-null males do not require CG17575 or seminase from a second male to bind SP from that male.
Cartoon: Pictorial representation of the experimental cross (fly images from Biorender). Females mated first with SP-null; ProtB-eGFP males [cyan sperm-head; DAPI(blue)+eGFP(green)] and then a day …
Figure 6—figure supplement 1
Western blot probed for seminase and CG17575.
Lanes/samples are 1: Fv, reproductive tract (RT) of three virgin females (negative control); 2: M, one pair of male accessory glands (positive control); 3–4: RT of five females mated to wild type …
Tables
Key resources table
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Genetic reagent (D. melanogaster) | Tudor | R. Boswell; similar stock now available from BloomingtonDrosophila Stock Center | BDSC:1735; FBst0001735;RRID:BDSC_1735 | FlyBase Genotype:tud1 bw1 sp1/CyO |
| Genetic reagent (D. melanogaster) | Δ325/TM3; Sb ry (SP-knockout line) | Gift from Eric Kubli | ||
| Genetic reagent (D. melanogaster) | Δ130/TM3; Sb ry (deficiency line) | Gift from Eric Kubli | ||
| Genetic reagent (D. melanogaster) | ProtB-eGFP(X); TM3/TM6 | Gift from Scott Pitnick | ||
| Genetic reagent (D. melanogaster) | ProtB-DsRed | Gift from Scott Pitnick | ||
| Antibody | anti-SP (rabbit polyclonal) | Wolfner lab | IF (1:200), WB (1:2000) | |
| Antibody | anti-CG1656 (rabbit polyclonal) | Wolfner lab | IF (1:100), WB (1:1000) | |
| Antibody | anti-CG1652 (rabbit polyclonal) | Wolfner lab | IF (1:50), WB (1:500) | |
| Antibody | anti-CG9997 (rabbit polyclonal) | Wolfner lab | IF (1:50), WB (1:1000) | |
| Antibody | IgG (H+L) Goat anti-Rabbit, Alexa Fluor 488 (goat anti-rabbit polyclonal) | Invitrogen | Cat. # A11008 RRID:AB_143165 | IF (1:300) |
| Antibody | IgG (H+L) Goat anti-Rabbit, Alexa Fluor 594 (goat anti-rabbit polyclonal) | Invitrogen | Cat. # A11012 RRID:AB_2534079 | IF (1:300) |
| Antibody | anti-Antares (rabbit polyclonal) | Wolfner lab | WB (1:500) | |
| Antibody | anti-seminase (rabbit polyclonal) | Wolfner lab | WB (1:1000) | |
| Antibody | anti-CG17575 (rabbit polyclonal) | Wolfner lab | WB (1:1000) | |
| Antibody | Anti-actin (mouse monoclonal) | Millipore Corp | Cat# MAB1501 RRID:AB_2223041 | WB (1:3000) |
| Antibody | Peroxidase AffiniPure Goat Anti-Rabbit IgG (goat anti-rabbit polyclonal) | Jackson Research | Code#111-035-003 RRID:AB_2313567 | WB (1:2000) |
| Antibody | Peroxidase AffiniPure Goat Anti-Mouse IgG (goat anti-mouse polyclonal) | Jackson Research | Code#115-035-003 RRID:AB_10015289 | WB (1:2000) |
| Other | DAPI stain | Invitrogen | Cat. # PI62247 | (1 µg/mL) |
| Other | Poly-L-Lysine (0.1 % w/v in H2O) | Sigma | P8920-100ML | 0.01% w/v in H2O |
| Other | Albumin from Bovine Serum (BSA) | Sigma | A9418-50G | 5% in 1X PBS |
| Other | CitiFluor Mountant Solution | Electron Microscopy Sciences | Cat. #17970–100 | |
| Software, Algorithm | Graph Pad Prism | RRID:SCR_002798 | Version 6.01 |
