A live cell reporter of exosome secretion and uptake reveals pathfinding behavior of migrating cells

Bong Hwan Sung, Ariana Lersner, Jorge Guerrero, Evan S. Krystofiak, David Inman, Roxanne Pelletier, Andries Zijlstra, Suzanne M. Ponik and Alissa M. Weaver ()
Additional contact information
Bong Hwan Sung: Vanderbilt University School of Medicine
Ariana Lersner: Vanderbilt University School of Medicine
Jorge Guerrero: University of Wisconsin-Madison
Evan S. Krystofiak: Vanderbilt University Cell Imaging Shared Resource
David Inman: University of Wisconsin-Madison
Roxanne Pelletier: Vanderbilt University School of Medicine
Andries Zijlstra: Vanderbilt University School of Medicine
Suzanne M. Ponik: University of Wisconsin-Madison
Alissa M. Weaver: Vanderbilt University School of Medicine

Nature Communications, 2020, vol. 11, issue 1, 1-15

Abstract: Abstract Small extracellular vesicles called exosomes affect multiple autocrine and paracrine cellular phenotypes. Understanding the function of exosomes requires a variety of tools, including live imaging. Our previous live-cell reporter, pHluorin-CD63, allows dynamic subcellular monitoring of exosome secretion in migrating and spreading cells. However, dim fluorescence and the inability to make stably-expressing cell lines limit its use. We incorporated a stabilizing mutation in the pHluorin moiety, M153R, which now exhibits higher, stable expression in cells and superior monitoring of exosome secretion. Using this improved construct, we visualize secreted exosomes in 3D culture and in vivo and identify a role for exosomes in promoting leader–follower behavior in 2D and 3D migration. Incorporating an additional non-pH-sensitive red fluorescent tag allows visualization of the exosome lifecycle, including multivesicular body (MVB) trafficking, MVB fusion, exosome uptake and endosome acidification. This reporter will be a useful tool for understanding both autocrine and paracrine roles of exosomes.

Date: 2020
References: Add references at CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41467-020-15747-2 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-15747-2

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-020-15747-2

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().