Analysis of Yeast Extracellular Vesicles

  • Marcio L. Rodrigues
  • Debora L. Oliveira
  • Gabriele Vargas
  • Wendell Girard-Dias
  • Anderson J. Franzen
  • Susana Frasés
  • Kildare Miranda
  • Leonardo Nimrichter
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1459)

Abstract

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.

Key words

Fungi Yeast Extracellular vesicles Cryptococcus Candida Saccharomyces 

Notes

Acknowledgement

We are grateful to Jorge José-Jó Bastos Ferreira for helpful suggestions. This work was supported by grants from the Brazilian agencies CNPq, CAPES, and FAPERJ. The authors also acknowledge support from the Instituto Nacional de Ciência e Tecnologia de Inovação em Doenças Negligenciadas (INCT-IDN). M.L.R. is the recipient of a Pathfinder Award from the Wellcome Trust (UK).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Marcio L. Rodrigues
    • 1
    • 2
  • Debora L. Oliveira
    • 1
  • Gabriele Vargas
    • 2
  • Wendell Girard-Dias
    • 3
    • 4
  • Anderson J. Franzen
    • 5
  • Susana Frasés
    • 4
    • 6
  • Kildare Miranda
    • 4
  • Leonardo Nimrichter
    • 2
  1. 1.Fundação Oswaldo Cruz (Fiocruz)Centro de Desenvolvimento Tecnológico em Saúde (CDTS) Fundação Oswaldo CruzRio de JaneiroBrazil
  2. 2.Instituto de Microbiologia Professor Paulo de GóesUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil
  3. 3.Instituto Oswaldo CruzFiocruz, Rio de JaneiroBrazil
  4. 4.Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho and Instituto Nacional de Ciência e Tecnologia em Biologia Estrutural e BioimagemUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil
  5. 5.Laboratório de Tecnologia em Cultura de CélulasCentro Universitário Estadual da Zona OesteRio de JaneiroBrazil
  6. 6.Laboratório de Biologia Estrutural, Diretoria de Programas, Instituto Nacional de MetrologiaNormalização e Qualidade Industrial (INMETRO)Xerém, Rio de JaneiroBrazil

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