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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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Rab GTPase signatures define different SC subcellular compartments. (A–D) SCs from 3-d-old males expressing different Rab-YFP constructs and stained with LysoTracker red. SCs have 5–10 small acidic Rab7-YFP–positive iLEs (A, arrowheads; most of these are usually located more apically than this confocal section; see Fig. 3) and 2.4 ± 0.9 (n = 11) Rab7-YFP–positive large vacuoles, 85% of which are acidic mMVBLs (A, asterisks; similar numbers [2.9 ± 0.5, n = 9] of large acidic compartments are seen in CD63-GFP–expressing SCs at this stage). Other Rab-YFP lines reveal Rab11-YFP–positive, nonacidic SVs (B, +; and D), many Rab5-YFP–positive small nonacidic compartments (C, arrowheads), and more rarely (4/16 SCs), a Rab5-YFP–positive small acidic compartment (C, arrows). In some fixed tub>Rab11-YFP SCs stained for Rab7, Rab7 colocalizes with Rab11-YFP in parts of the limiting membrane (D, arrow) of a single SV (D, #). DAPI stains nuclei (blue). Approximate outline of SC is marked in all images. Bars, 5 µm.
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fig2: Rab GTPase signatures define different SC subcellular compartments. (A–D) SCs from 3-d-old males expressing different Rab-YFP constructs and stained with LysoTracker red. SCs have 5–10 small acidic Rab7-YFP–positive iLEs (A, arrowheads; most of these are usually located more apically than this confocal section; see Fig. 3) and 2.4 ± 0.9 (n = 11) Rab7-YFP–positive large vacuoles, 85% of which are acidic mMVBLs (A, asterisks; similar numbers [2.9 ± 0.5, n = 9] of large acidic compartments are seen in CD63-GFP–expressing SCs at this stage). Other Rab-YFP lines reveal Rab11-YFP–positive, nonacidic SVs (B, +; and D), many Rab5-YFP–positive small nonacidic compartments (C, arrowheads), and more rarely (4/16 SCs), a Rab5-YFP–positive small acidic compartment (C, arrows). In some fixed tub>Rab11-YFP SCs stained for Rab7, Rab7 colocalizes with Rab11-YFP in parts of the limiting membrane (D, arrow) of a single SV (D, #). DAPI stains nuclei (blue). Approximate outline of SC is marked in all images. Bars, 5 µm.

Mentions: We analyzed the large SC compartments in more detail using LysoTracker red–stained living glands. Counting vacuoles by scanning through multiple focal planes revealed remarkably consistent numbers of large (>2 µm) and small (<2 µm) CD63-positive acidic compartments and large nonacidic CD63-positive compartments in each SC of 6-d-old adults (Fig. 1 I). Both classes of acidic compartments were also visible in non–CD63-GFP–expressing SCs (Figs. S1, G versus H; and 2, A–C) and therefore were not induced by CD63-GFP expression. We hypothesized that the smaller apical acidic compartments were immature late endosomes (iLEs; CD63-GFP+/LysoTracker+ and <2 µm; Fig. 1 I), and the large acidic compartments, at least one of which contained small CD63-GFP–positive puncta, were unusually large (≤10 µm in 6-d-old SCs) maturing MVBs or lysosomes (mMVBLs; CD63-GFP+/LysoTracker+ and >2 µm; Fig. 1 I). In addition, some, but not all, nonacidic SV compartments (CD63-GFP+/Lysotracker− and >2 µm; Fig. 1 I) contained larger CD63-GFP–positive intraluminal structures.


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)

Rab GTPase signatures define different SC subcellular compartments. (A–D) SCs from 3-d-old males expressing different Rab-YFP constructs and stained with LysoTracker red. SCs have 5–10 small acidic Rab7-YFP–positive iLEs (A, arrowheads; most of these are usually located more apically than this confocal section; see Fig. 3) and 2.4 ± 0.9 (n = 11) Rab7-YFP–positive large vacuoles, 85% of which are acidic mMVBLs (A, asterisks; similar numbers [2.9 ± 0.5, n = 9] of large acidic compartments are seen in CD63-GFP–expressing SCs at this stage). Other Rab-YFP lines reveal Rab11-YFP–positive, nonacidic SVs (B, +; and D), many Rab5-YFP–positive small nonacidic compartments (C, arrowheads), and more rarely (4/16 SCs), a Rab5-YFP–positive small acidic compartment (C, arrows). In some fixed tub>Rab11-YFP SCs stained for Rab7, Rab7 colocalizes with Rab11-YFP in parts of the limiting membrane (D, arrow) of a single SV (D, #). DAPI stains nuclei (blue). Approximate outline of SC is marked in all images. Bars, 5 µm.
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Related In: Results  -  Collection

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fig2: Rab GTPase signatures define different SC subcellular compartments. (A–D) SCs from 3-d-old males expressing different Rab-YFP constructs and stained with LysoTracker red. SCs have 5–10 small acidic Rab7-YFP–positive iLEs (A, arrowheads; most of these are usually located more apically than this confocal section; see Fig. 3) and 2.4 ± 0.9 (n = 11) Rab7-YFP–positive large vacuoles, 85% of which are acidic mMVBLs (A, asterisks; similar numbers [2.9 ± 0.5, n = 9] of large acidic compartments are seen in CD63-GFP–expressing SCs at this stage). Other Rab-YFP lines reveal Rab11-YFP–positive, nonacidic SVs (B, +; and D), many Rab5-YFP–positive small nonacidic compartments (C, arrowheads), and more rarely (4/16 SCs), a Rab5-YFP–positive small acidic compartment (C, arrows). In some fixed tub>Rab11-YFP SCs stained for Rab7, Rab7 colocalizes with Rab11-YFP in parts of the limiting membrane (D, arrow) of a single SV (D, #). DAPI stains nuclei (blue). Approximate outline of SC is marked in all images. Bars, 5 µm.
Mentions: We analyzed the large SC compartments in more detail using LysoTracker red–stained living glands. Counting vacuoles by scanning through multiple focal planes revealed remarkably consistent numbers of large (>2 µm) and small (<2 µm) CD63-positive acidic compartments and large nonacidic CD63-positive compartments in each SC of 6-d-old adults (Fig. 1 I). Both classes of acidic compartments were also visible in non–CD63-GFP–expressing SCs (Figs. S1, G versus H; and 2, A–C) and therefore were not induced by CD63-GFP expression. We hypothesized that the smaller apical acidic compartments were immature late endosomes (iLEs; CD63-GFP+/LysoTracker+ and <2 µm; Fig. 1 I), and the large acidic compartments, at least one of which contained small CD63-GFP–positive puncta, were unusually large (≤10 µm in 6-d-old SCs) maturing MVBs or lysosomes (mMVBLs; CD63-GFP+/LysoTracker+ and >2 µm; Fig. 1 I). In addition, some, but not all, nonacidic SV compartments (CD63-GFP+/Lysotracker− and >2 µm; Fig. 1 I) contained larger CD63-GFP–positive intraluminal structures.

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