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Visualization of Extracellular Vesicles of Prokaryotes and Eukaryotic Microbes
Applied Microscopy 2018;48:96-101
Published online December 28, 2018
© 2018 Korean Society of Microscopy.

Ki Woo Kim

1School of Ecology and Environmental System, Kyungpook National University, Sangju 37224, Korea, 2Tree Diagnostic Center, Kyungpook National University, Sangju 37224, Korea
Correspondence to: *Correspondence to: Kim KW, http://orcid.org/0000-0002-7010-0336, Tel: +82-54-530-1246, Fax: +82-54-530-1248, E-mail: kiwoo@knu.ac.kr
Received December 19, 2018; Revised December 27, 2018; Accepted December 27, 2018.
his is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract

The release of nanoscale membrane-bound vesicles is common in all three domains of life. These vesicles are involved in a variety of biological processes such as cell-to-cell communication, horizontal gene transfer, and substrate transport. Prokaryotes including bacteria and archaea release membrane vesicles (MVs) (20 to 400 nm in diameter) into their extracellular milieu. In spite of structural differences in cell envelope, both Gram-positive and negative bacteria produce MVs that contain the cell membrane of each bacterial species. Archaeal MVs characteristically show surface-layer encircling the vesicles. Filamentous fungi and yeasts as eukaryotic microbes produce bilayered exosomes that have varying electron density. Microbes also form intracellular vesicles and minicells that are similar to MVs and exosomes in shape. Electron and fluorescence microscopy could reveal the presence of DNA in MVs and exosomes. Given the biogenesis of extracellular vesicles from the donor cell, in situ high-resolution microscopy can provide insights on the structural mechanisms underlying the formation and release of microbial extracellular vesicles.

Keywords : Archaea, Bacteria, Fungi, Membrane vesicle
Figures
Fig. 1. Schematics of extracellular vesicles of microbial cells. ERG, ergosterol; GlcConj, glycoconjugates; GSL, glycosphingolipids; LPS, lipopolysaccharide; OMP, outer membrane protein. From with permission from the publisher.
Fig. 2. Membrane vesicles of Gram-negative bacteria. (A) Transmission electron micrograph of Escherichia coli (b) labeled with anti-lipopolysaccharide antibody and Protein A Gold. Subsets depict membrane vesicles. Bar=150 nm. (B) Scanning electron micrograph of Xylella fastidiosa having membrane vesicles (arrows). Bar=200 nm. A: from ; B: from with permission from the publisher.
Fig. 3. Transmission electron micrographs of membrane vesicles of Staphylococcus aureus. (A) Membrane vesicles (arrow) released from the cell. (B) Membrane vesicles pelleted by ultracentrifugation from the bacterial culture. Bars=100 nm. A and B: from with permission from the publisher.
Fig. 4. Transmission electron micrograph of membrane vesicles of Sulfolobus solfataricus. These vesicles are surrounded with surface-layer (arrow). From with permission from the publisher.
Fig. 5. Transmission electron micrographs of membrane vesicles of Saccharomyces cerevisiae. (A–D) Exosomes. They are bilayered structures with varying electron density. Bar=100 nm. A–D: from with permission from the publisher.
Fig. 6. Transmission electron micrographs of Agrobacterium tumefaciens. (A) Cell having the partially filled acidocalcisome (arrow). (B) Cell having the empty acidocalcisome (arrow). A and B: from with permission from the publisher.
Fig. 7. Localization of DNA in membrane vesicles. (A–C) Super resolution micrographs of membrane vesicles of Pseudomonas aeruginosa. (A) Membrane vesicles stained with the lipophilic fluorescent dye, DiO (green). Bar=1 μm. (B) Membrane vesicles stained with the DNA-specific dye, 5- ethynyl-2-deoxuridine (EdU). (C) Merged image of (A) and (B). (D) Transmission electron micrograph of membrane vesicles of Shewanella vesiculosa. Electron-dense particles indicate gold particles with an antibody specific for DNA. OM, outer membrane; PM, plasma membrane. A–C: from ; D: from with permission from the publisher.
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December 2018, 48 (4)
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