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BMP-regulated exosomes from Drosophila male reproductive glands reprogram female behavior.

Corrigan L, Redhai S, Leiblich A, Fan SJ, Perera SM, Patel R, Gandy C, Wainwright SM, Morris JF, Hamdy F, Goberdhan DC, Wilson C - J. Cell Biol. (2014)

Bottom Line: Male reproductive glands secrete signals into seminal fluid to facilitate reproductive success.Exosome release was required to inhibit female remating behavior, suggesting that exosomes are downstream effectors of BMP signaling.These results demonstrate a new function for the MVB-exosome pathway in the reproductive tract that appears to be conserved across evolution.

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Affiliation: Department of Physiology, Anatomy and Genetics and Nuffield Department of Surgical Sciences, University of Oxford, Oxford OX1 3QX, England, UK.

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SC exosomes interact with sperm in females. (A) Schematic of female reproductive tract in D showing mating plug (mp), uterus (ut), oviduct (ovi), and sperm storage organs (spermathecae [spt] and seminal receptacle [sr]) as well as the autofluorescent hindgut (HG). (B and C) w1118 female reproductive tract dissected 20 min after mating to either a control w1118 (B) or SC>CD63-GFP (C) male, imaged under identical confocal settings. Images show lumen of the female uterus (asterisks) containing sperm whose heads are visible with DAPI staining (arrowheads). (D and E) Female reproductive tracts 20 min after mating to a protamineB-RFP; SC>CD63-GFP male. (D) ProtamineB-RFP (prot-RFP), which marks sperm heads, accumulates in the anterior uterus (red arrowhead) and oviduct (red arrow). Most CD63-GFP–positive exosomes localize to the posterior uterus (green asterisk), but they can also be seen in the anterior uterus (green arrowhead) and oviduct (green arrow). The autofluorescent mating plug (closed asterisk) is shown. (E) CD63-GFP–positive exosomes colocalize with sperm heads marked by protamineB-RFP (open arrows). (F) Orthogonal view of a z stack through near-complete ring of CD63-GFP fused to ProtamineB-RFP sperm head (arrows). The confocal image below the blue line shows part of the female reproductive tract lumen. The two images above the blue line, separated by a white line, are green (top) and red/green (bottom) z stacks captured in the z plane marked in the bottom image. Note multiple pairs of parallel GFP-positive lines (arrowheads), potentially produced by sperm tail fusion. (G) These (arrowheads) and irregular extended lines of fluorescence (arrow) are also seen in G, which shows a maximum 3D projection image of a z stack from a female anterior uterus and oviduct where the epithelium is marked by actin>CD8-RFP (also shown in Video 3). (H) Female reproductive tracts were dissected and fixed at specific times after the start of mating to SC>CD63-GFP males. The frequency of exosome–sperm interaction events within the female reproductive tract was analyzed (n = 8 for each time point after the start of mating [ASM]). Only fusions to the sperm heads are included. Bars: (B and C) 50 µm; (D) 200 µm; (E and F) 5 µm; (G) 20 µm.
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fig6: SC exosomes interact with sperm in females. (A) Schematic of female reproductive tract in D showing mating plug (mp), uterus (ut), oviduct (ovi), and sperm storage organs (spermathecae [spt] and seminal receptacle [sr]) as well as the autofluorescent hindgut (HG). (B and C) w1118 female reproductive tract dissected 20 min after mating to either a control w1118 (B) or SC>CD63-GFP (C) male, imaged under identical confocal settings. Images show lumen of the female uterus (asterisks) containing sperm whose heads are visible with DAPI staining (arrowheads). (D and E) Female reproductive tracts 20 min after mating to a protamineB-RFP; SC>CD63-GFP male. (D) ProtamineB-RFP (prot-RFP), which marks sperm heads, accumulates in the anterior uterus (red arrowhead) and oviduct (red arrow). Most CD63-GFP–positive exosomes localize to the posterior uterus (green asterisk), but they can also be seen in the anterior uterus (green arrowhead) and oviduct (green arrow). The autofluorescent mating plug (closed asterisk) is shown. (E) CD63-GFP–positive exosomes colocalize with sperm heads marked by protamineB-RFP (open arrows). (F) Orthogonal view of a z stack through near-complete ring of CD63-GFP fused to ProtamineB-RFP sperm head (arrows). The confocal image below the blue line shows part of the female reproductive tract lumen. The two images above the blue line, separated by a white line, are green (top) and red/green (bottom) z stacks captured in the z plane marked in the bottom image. Note multiple pairs of parallel GFP-positive lines (arrowheads), potentially produced by sperm tail fusion. (G) These (arrowheads) and irregular extended lines of fluorescence (arrow) are also seen in G, which shows a maximum 3D projection image of a z stack from a female anterior uterus and oviduct where the epithelium is marked by actin>CD8-RFP (also shown in Video 3). (H) Female reproductive tracts were dissected and fixed at specific times after the start of mating to SC>CD63-GFP males. The frequency of exosome–sperm interaction events within the female reproductive tract was analyzed (n = 8 for each time point after the start of mating [ASM]). Only fusions to the sperm heads are included. Bars: (B and C) 50 µm; (D) 200 µm; (E and F) 5 µm; (G) 20 µm.

Mentions: We investigated whether the interaction between SC exosomes and target cells primarily takes place after mating. SC>CD63-GFP males transferred fluorescent puncta into the lumen of the female reproductive tract during mating (Fig. 6, A–D). Multiply mated males of 11 d and older can sporadically transfer SCs to females (Leiblich et al., 2012). Importantly, all mating experiments performed in our current study were with 4-d-old males; we never observed transfer of fluorescent SCs in >50 mated females that we imaged, indicating that SC transfer is not involved in the phenotypes observed.


BMP-regulated exosomes from Drosophila male reproductive glands reprogram female behavior.

Corrigan L, Redhai S, Leiblich A, Fan SJ, Perera SM, Patel R, Gandy C, Wainwright SM, Morris JF, Hamdy F, Goberdhan DC, Wilson C - J. Cell Biol. (2014)

SC exosomes interact with sperm in females. (A) Schematic of female reproductive tract in D showing mating plug (mp), uterus (ut), oviduct (ovi), and sperm storage organs (spermathecae [spt] and seminal receptacle [sr]) as well as the autofluorescent hindgut (HG). (B and C) w1118 female reproductive tract dissected 20 min after mating to either a control w1118 (B) or SC>CD63-GFP (C) male, imaged under identical confocal settings. Images show lumen of the female uterus (asterisks) containing sperm whose heads are visible with DAPI staining (arrowheads). (D and E) Female reproductive tracts 20 min after mating to a protamineB-RFP; SC>CD63-GFP male. (D) ProtamineB-RFP (prot-RFP), which marks sperm heads, accumulates in the anterior uterus (red arrowhead) and oviduct (red arrow). Most CD63-GFP–positive exosomes localize to the posterior uterus (green asterisk), but they can also be seen in the anterior uterus (green arrowhead) and oviduct (green arrow). The autofluorescent mating plug (closed asterisk) is shown. (E) CD63-GFP–positive exosomes colocalize with sperm heads marked by protamineB-RFP (open arrows). (F) Orthogonal view of a z stack through near-complete ring of CD63-GFP fused to ProtamineB-RFP sperm head (arrows). The confocal image below the blue line shows part of the female reproductive tract lumen. The two images above the blue line, separated by a white line, are green (top) and red/green (bottom) z stacks captured in the z plane marked in the bottom image. Note multiple pairs of parallel GFP-positive lines (arrowheads), potentially produced by sperm tail fusion. (G) These (arrowheads) and irregular extended lines of fluorescence (arrow) are also seen in G, which shows a maximum 3D projection image of a z stack from a female anterior uterus and oviduct where the epithelium is marked by actin>CD8-RFP (also shown in Video 3). (H) Female reproductive tracts were dissected and fixed at specific times after the start of mating to SC>CD63-GFP males. The frequency of exosome–sperm interaction events within the female reproductive tract was analyzed (n = 8 for each time point after the start of mating [ASM]). Only fusions to the sperm heads are included. Bars: (B and C) 50 µm; (D) 200 µm; (E and F) 5 µm; (G) 20 µm.
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fig6: SC exosomes interact with sperm in females. (A) Schematic of female reproductive tract in D showing mating plug (mp), uterus (ut), oviduct (ovi), and sperm storage organs (spermathecae [spt] and seminal receptacle [sr]) as well as the autofluorescent hindgut (HG). (B and C) w1118 female reproductive tract dissected 20 min after mating to either a control w1118 (B) or SC>CD63-GFP (C) male, imaged under identical confocal settings. Images show lumen of the female uterus (asterisks) containing sperm whose heads are visible with DAPI staining (arrowheads). (D and E) Female reproductive tracts 20 min after mating to a protamineB-RFP; SC>CD63-GFP male. (D) ProtamineB-RFP (prot-RFP), which marks sperm heads, accumulates in the anterior uterus (red arrowhead) and oviduct (red arrow). Most CD63-GFP–positive exosomes localize to the posterior uterus (green asterisk), but they can also be seen in the anterior uterus (green arrowhead) and oviduct (green arrow). The autofluorescent mating plug (closed asterisk) is shown. (E) CD63-GFP–positive exosomes colocalize with sperm heads marked by protamineB-RFP (open arrows). (F) Orthogonal view of a z stack through near-complete ring of CD63-GFP fused to ProtamineB-RFP sperm head (arrows). The confocal image below the blue line shows part of the female reproductive tract lumen. The two images above the blue line, separated by a white line, are green (top) and red/green (bottom) z stacks captured in the z plane marked in the bottom image. Note multiple pairs of parallel GFP-positive lines (arrowheads), potentially produced by sperm tail fusion. (G) These (arrowheads) and irregular extended lines of fluorescence (arrow) are also seen in G, which shows a maximum 3D projection image of a z stack from a female anterior uterus and oviduct where the epithelium is marked by actin>CD8-RFP (also shown in Video 3). (H) Female reproductive tracts were dissected and fixed at specific times after the start of mating to SC>CD63-GFP males. The frequency of exosome–sperm interaction events within the female reproductive tract was analyzed (n = 8 for each time point after the start of mating [ASM]). Only fusions to the sperm heads are included. Bars: (B and C) 50 µm; (D) 200 µm; (E and F) 5 µm; (G) 20 µm.
Mentions: We investigated whether the interaction between SC exosomes and target cells primarily takes place after mating. SC>CD63-GFP males transferred fluorescent puncta into the lumen of the female reproductive tract during mating (Fig. 6, A–D). Multiply mated males of 11 d and older can sporadically transfer SCs to females (Leiblich et al., 2012). Importantly, all mating experiments performed in our current study were with 4-d-old males; we never observed transfer of fluorescent SCs in >50 mated females that we imaged, indicating that SC transfer is not involved in the phenotypes observed.

Bottom Line: Male reproductive glands secrete signals into seminal fluid to facilitate reproductive success.Exosome release was required to inhibit female remating behavior, suggesting that exosomes are downstream effectors of BMP signaling.These results demonstrate a new function for the MVB-exosome pathway in the reproductive tract that appears to be conserved across evolution.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Physiology, Anatomy and Genetics and Nuffield Department of Surgical Sciences, University of Oxford, Oxford OX1 3QX, England, UK.

Show MeSH
Related in: MedlinePlus