Analysis of Yeast Extracellular Vesicles

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1459; p. 175
• Main Authors: Rodrigues, Marcio L, Oliveira, Debora L, Vargas, Gabriele, Girard-Dias, Wendell, Franzen, Anderson J, Frasés, Susana, Miranda, Kildare, Nimrichter, Leonardo
• Format: Journal Article
• Language: English
• Published: United States 2016
• Subjects: Animals; Cell Fractionation; Cell Line; Extracellular Vesicles - metabolism; Extracellular Vesicles - ultrastructure; Fungal Proteins - metabolism; Lipid Metabolism; Mice; Secretory Vesicles - metabolism; Transport Vesicles - metabolism; Yeasts - metabolism; Fungi; Cryptococcus; Extracellular vesicles; Yeast; Saccharomyces; Candida
• ISSN: 1940-6029
• DOI: 10.1007/978-1-4939-3804-9_12

Extracellular vesicles (EV) are important carriers of biologically active components in a number of organisms, including fungal cells. Experimental characterization of fungal EVs suggested that these membranous compartments are likely involved in the regulation of several biological events. In fungal pathogens, these events include mechanisms of disease progression and/or control, suggesting potential targets for therapeutic intervention or disease prophylaxis. In this manuscript we describe methods that have been used in the last 10 years for the characterization of EVs produced by yeast forms of several fungal species. Experimental approaches detailed in this chapter include ultracentrifugation methods for EV fractionation, chromatographic approaches for analysis of EV lipids, microscopy techniques for analysis of both intracellular and extracellular vesicular compartments, interaction of EVs with host cells, and physical chemical analysis of EVs by dynamic light scattering.