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      Rab‐mediated trafficking in the secondary cells of Drosophila male accessory glands and its role in fecundity

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          Abstract

          The male seminal fluid contains factors that affect female post‐mating behavior and physiology. In Drosophila, most of these factors are secreted by the two epithelial cell types that make up the male accessory gland: the main and secondary cells. Although secondary cells represent only ~4% of the cells of the accessory gland, their contribution to the male seminal fluid is essential for sustaining the female post‐mating response. To better understand the function of the secondary cells, we investigated their molecular organization, particularly with respect to the intracellular membrane transport machinery. We determined that large vacuole‐like structures found in the secondary cells are trafficking hubs labeled by Rab6, 7, 11 and 19. Furthermore, these organelles require Rab6 for their formation and many are essential in the process of creating the long‐term postmating behavior of females. In order to better serve the intracellular membrane and protein trafficking communities, we have created a searchable, online, open‐access imaging resource to display our complete findings regarding Rab localization in the accessory gland.

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          Most cited references74

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          Rab7: a key to lysosome biogenesis.

          The molecular machinery behind lysosome biogenesis and the maintenance of the perinuclear aggregate of late endocytic structures is not well understood. A likely candidate for being part of this machinery is the small GTPase Rab7, but it is unclear whether this protein is associated with lysosomes or plays any role in the regulation of the perinuclear lysosome compartment. Previously, Rab7 has mainly been implicated in transport from early to late endosomes. We have now used a new approach to analyze the role of Rab7: transient expression of Enhanced Green Fluorescent Protein (EGFP)-tagged Rab7 wt and mutant proteins in HeLa cells. EGFP-Rab7 wt was associated with late endocytic structures, mainly lysosomes, which aggregated and fused in the perinuclear region. The size of the individual lysosomes as well as the degree of perinuclear aggregation increased with the expression levels of EGFP-Rab7 wt and, more dramatically, the active EGFP-Rab7Q67L mutant. In contrast, upon expression of the dominant-negative mutants EGFP-Rab7T22N and EGFP-Rab7N125I, which localized mainly to the cytosol, the perinuclear lysosome aggregate disappeared and lysosomes, identified by colocalization of cathepsin D and lysosome-associated membrane protein-1, became dispersed throughout the cytoplasm, they were inaccessible to endocytosed molecules such as low-density lipoprotein, and their acidity was strongly reduced, as determined by decreased accumulation of the acidotropic probe LysoTracker Red. In contrast, early endosomes associated with Rab5 and the transferrin receptor, late endosomes enriched in the cation-independent mannose 6-phosphate receptor, and the trans-Golgi network, identified by its enrichment in TGN-38, were unchanged. These data demonstrate for the first time that Rab7, controlling aggregation and fusion of late endocytic structures/lysosomes, is essential for maintenance of the perinuclear lysosome compartment.
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            Sex peptide causes mating costs in female Drosophila melanogaster.

            Conflicts between females and males over reproductive decisions are common . In Drosophila, as in many other organisms, there is often a conflict over how often to mate. The mating frequency that maximizes male reproductive success is higher than that which maximizes female reproductive success . In addition, frequent mating reduces female lifespan and reproductive success , a cost that is mediated by male ejaculate accessory gland proteins (Acps) . We demonstrate here that a single Acp, the sex peptide (SP or Acp70A), which decreases female receptivity and stimulates egg production in the first matings of virgin females , is a major contributor to Acp-mediated mating costs in females. Females continuously exposed to SP-deficient males (which produce no detectable SP ) had significantly higher fitness and higher lifetime reproductive success than control females. Hence, rather than benefiting both sexes, receipt of SP decreases female fitness, making SP the first identified gene that is likely to play a central role in sexual conflict.
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              Organization of vesicular trafficking in epithelia.

              Experiments using mammalian epithelial cell lines have elucidated biosynthetic and recycling pathways for apical and basolateral plasma-membrane proteins, and have identified components that guide apical and basolateral proteins along these pathways. These components include apical and basolateral sorting signals, adaptors for basolateral signals, and docking and fusion proteins for vesicular trafficking. Recent live-cell-imaging studies provide a real-time view of sorting processes in epithelial cells, including key roles for actin, microtubules and motors in the organization of post-Golgi trafficking.
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                Author and article information

                Contributors
                francois.karch@unige.ch
                marko.brankatschk@tu-dresden.de
                robert.maeda@unige.ch
                Journal
                Traffic
                Traffic
                10.1111/(ISSN)1600-0854
                TRA
                Traffic (Copenhagen, Denmark)
                John Wiley & Sons A/S (Former Munksgaard )
                1398-9219
                1600-0854
                26 December 2018
                February 2019
                : 20
                : 2 ( doiID: 10.1111/tra.2019.20.issue-2 )
                : 137-151
                Affiliations
                [ 1 ] Department of Genetics and Evolution, Section of Biology Sciences Faculty, University of Geneva Geneva Switzerland
                [ 2 ] Department of Physiology, Anatomy and Genetics University of Oxford Oxford UK
                [ 3 ] Biotechnology Center of the TU Dresden Dresden Germany
                [ 4 ] Max Planck Institute of Molecular Cell Biology and Genetics Dresden Germany
                Author notes
                [*] [* ] Correspondence

                Robert K. Maeda, Department of Genetics and Evolution, Section of Biology, Sciences Faculty, University of Geneva, Geneva, Switzerland.

                Email: robert.maeda@ 123456unige.ch

                Marko Brankatschk, Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.

                Email: marko.brankatschk@ 123456tu-dresden.de

                François Karch, Department of Genetics and Evolution, Section of Biology, Sciences Faculty, University of Geneva, Geneva, Switzerland.

                Email: francois.karch@ 123456unige.ch

                Author information
                https://orcid.org/0000-0001-6555-1215
                Article
                TRA12622
                10.1111/tra.12622
                6492190
                30426623
                0324739b-8f77-4932-93bd-38e1669175f9
                © 2018 The Authors. Traffic published by John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 29 March 2018
                : 01 November 2018
                : 06 November 2018
                Page count
                Figures: 7, Tables: 0, Pages: 16, Words: 10553
                Funding
                Funded by: Biotechnology and Biological Sciences Research Council
                Award ID: BB/K017462/1
                Funded by: Deutsche Forschungsgemeinschaft
                Award ID: DFG: BR5490/2
                Funded by: Donation Claraz
                Funded by: Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung
                Award ID: 31003A_170179
                Funded by: State of Geneva
                Funded by: Welcome Trust (Strategic Award)
                Award ID: 102347/Z/13/Z
                Award ID: #091911
                Categories
                Original Article
                Original Articles
                Custom metadata
                2.0
                tra12622
                February 2019
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.6.2.1 mode:remove_FC converted:01.05.2019

                Sociology
                confocal microscopy,drosophila melanogaster,male accessory glands,postmating response,rab19,rab6,vacuole‐like compartments

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