Rey-Cadilhac F. auteur aut IRD Rachenne F. auteur aut IRD Marquant A. auteur aut IRD Him J. L. K. auteur aut IRD Ancelin A. auteur aut IRD Foisor V. auteur aut IRD Morille M. auteur aut IRD Lyonnais S. auteur aut IRD Cazevieille C. auteur aut IRD Missé Dorothée auteur aut IRD Pompon Julien auteur aut IRD text journalArticle eng text/pdf reformatted digital access Extracellular vesicles (EVs) are non-replicative, cell-derived membranous structures secreted by potentially all eukaryotic cells, playing a crucial role in intercellular communication. The study of EVs requires approaches and tools, which have predominantly been developed for mammalian models. Here, we undertook a multimodal characterization of mosquito EVs to provide a technical and knowledge foundation for their study. First, using a cell line model from Aedes aegypti and applying multiple analytical technologies (i.e., NTA, TEM, cryo-EM, and AFM), we observed that mosquito EVs range from 20 to 500 nm in diameter and that a majority are smaller than 100 nm. Second, we showed that smaller EVs are secreted in mosquito saliva. Third, we evaluated the capacity of differential centrifugation and size exclusion chromatography to separate mosquito EVs, revealing the strengths and weaknesses of each technology. Finally, we identified a mosquito homolog of CD63 as an extravesicular marker and the mosquito syntenin as a putative luminal marker. Overall, our results promote the development of tools and approaches for the study of mosquito EVs. specialized extracellular vesicles mosquito microscopy protein markers syntenin tetraspanin characterization analytical methods 020 052 13 1497795 [14 p.] 2025 2296-634X https://www.documentation.ird.fr/hor/fdi:010093447 10.3389/fcell.2025.1497795 2296-634X [F B010093447] https://www.documentation.ird.fr/hor/fdi:010093447 https://horizon.documentation.ird.fr/exl-doc/pleins_textes/2025-06/010093447.pdf IRD - Base Horizon / Pleins textes 2025-06-11 2025-11-04 fdi:010093447 fre