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Male-female communication enhances release of extracellular vesicles leading to high fertility in Drosophila

by NANOSENSORS
28 October 202215 March 2023
Science & Technology

Fig. 5 from “Male-female communication enhances release of extracellular vesicles leading to high fertility in Drosophila” by Javier Arturo Sanchez-Lopez et al.: The spermatheca releases EVs with specific characteristics. a–e The spermathecae (SSC > CD63-GFP) were cultured ex vivo to characterize the particles that were released to the spent media: a size distribution of CD63-GFP-positive particles (n = 25 spermathecae from 15 flies) using single-particle tracking of the Nanoimager. b Characterization of CD63-GFP particles by ExoView (100 spermathecae from 50-60 flies) using anti-GFP or anti-CD63. c Representative dSTORM image of EVs found in the spent media of SSC expressing CD63-GFP (n = 25 spermathecae from 15 flies), stained with AlexaFluor555 conjugated anti-CD63 primary antibody. The image shows CD63-GFP-positive EVs and single CD63 AlexaFluor555 molecules the EV’s surface. Scale bars = 1 µm and insets = 20 nm. d–e The morphology of EVs in the spent media was observed by e negative staining in STEM; scale bar = 200 nm; and by d Cryo-TEM; Scale bar = 100 nm (see also Supplementary Fig. 5a-c). f–i The spent media of ex vivo cultured spermathecae were analyzed for the presence of EVs: f Representative AFM scans of SSC-EVs isolated by acoustic sorting (n = 100 spermathecae from 70 flies), showing from left to right: height (nm), adhesion (pN) and Young’s modulus images (MPa). Scale bars = 100 nm (see Supplementary Fig. 5d–g). g–i Single-particle tracking in the nanoimager. g CellMaskTM-positive particle size distribution and concentration profiles of EVs in the spent media of spermathecae incubated in media alone (No ecdy) or with ecdysone (Ecdy) (see also Supplementary Movie 5 for a time lapse of spermathecae end apparatus and endosomal activity post-ecdysone stimulation and methods, ex vivo spermatheca culture section); media with only CellMaskTM stain served as a control for the formation of dye aggregates. h Particle concentration and i mean particle diameter (nm) from g; Box plots are the measurements of particles from four frames of spent media from 25 spermathecae; boxes represent maximum, median and minimum values with outliers; one-way ANOVA, with nonparametric Wilcoxon multiple comparison post-hoc test; *p

The female reproductive tract (female-RT) must decipher the repertoire of molecular cues received from the male during copulation in order to activate and coordinate tract… Read More »Male-female communication enhances release of extracellular vesicles leading to high fertility in Drosophila