@article{1edb4a37e8354c50952fd5d8e450b3e4,
title = "Characterization of extracellular vesicles produced by Aspergillus fumigatus protoplasts",
abstract = "Extracellular vesicles (EVs) are membranous compartments produced by yeast and mycelial forms of several fungal species. One of the difficulties in perceiving the role of EVs during the fungal life, and particularly in cell wall biogenesis, is caused by the presence of a thick cell wall. One alternative to have better access to these vesicles is to use protoplasts. This approach has been investigated here with Aspergillus fumigatus, one of the most common opportunistic fungal pathogens worldwide. Analysis of regenerating protoplasts by scanning electron microscopy and fluorescence microscopy indicated the occurrence of outer membrane projections in association with surface components and the release of particles with properties resembling those of fungal EVs. EVs in culture supernatants were characterized by transmission electron microscopy and nanoparticle tracking analysis. Proteomic and glycome analysis of EVs revealed the presence of a complex array of enzymes related to lipid/sugar metabolism, pathogenic processes, and cell wall biosynthesis. Our data indicate that (i) EV production is a common feature of different morphological stages of this major fungal pathogen and (ii) protoplastic EVs are promising tools for undertaking studies of vesicle functions in fungal cells.",
keywords = "Aspergillus, Aspergillus fumigatus, Conidia, Extracellular vesicles, Protoplasts",
author = "Juliana Rizzo and Thibault Chaze and Kildare Miranda and Roberson, {Robert W.} and Olivier Gorgette and Leonardo Nimrichter and Mariette Matondo and Latg{\'e}, {Jean Paul} and Anne Beauvais and Rodrigues, {Marcio L.}",
note = "Funding Information: M.L.R. was supported by grants from the Brazilian Ministry of Health (grant 440015/2018-9), Conselho Nacional de Desenvolvimento Cient{\'i}fico e Tecnol{\'o}gico (CNPq; grants 405520/2018-2 and 301304/2017-3), and Fiocruz (grants VPPCB-007-FIO-18 and VPPIS-001-FIO18). We also acknowledge support from the Instituto Nacional de Ci{\^e}ncia e Tecnologia de Inova{\c c}{\~a}o em Doen{\c c}as de Popula{\c c}{\~o}es Negligenciadas (INCT-IDPN). J.R. developed part of this work as a Ph.D. student supported by FAPERJ (E-26/201.991/2015) and is currently a postdoctoral fellow at Institut Pasteur (Paris, France), funded by the CAPES-Cofecub Program (88887.142840/2017-00). This research was partly supported by funds provided to J.-P.L. by the Laboratoire d'Excellence {"}Integrative Biology of Emerging Infectious Diseases{"} (grant ANR-10-LABX-62-IBEID), la Fondation pour la Recherche M{\'e}dicale (DEQ20150331722 LATGE Equipe FRM 2015). We are grateful to Leandro Honorato (Instituto de Microbiologia Paulo de Goes, UFRJ, Brazil) and Luna Sobrino Joffe (Stony Brook University, USA) for help with protoplast purification, Flavia Reis (Fiocruz, Brazil) for help with NTA, Fernando P. de Almeida for the help with superresolution microscopy (CENABIO/UFRJ, Brazil), and Vishukumar Aimanianda (Institut Pasteur, France) for discussions and for providing some of the antibodies for this study. Publisher Copyright: {\textcopyright} 2020 Rizzo et al.",
year = "2020",
doi = "10.1128/MSPHERE.00476-20",
language = "English (US)",
volume = "5",
journal = "mSphere",
issn = "2379-5042",
publisher = "American Society for Microbiology",
number = "4",
}