Dynamic imaging of the recycling endosomal network in macrophages

Wall, Adam A., Condon, Nicholas D., Yeo, Jeremy C., Hamilton, Nicholas A. and Stow, Jennifer L. (2015) Dynamic imaging of the recycling endosomal network in macrophages. Methods in Cell Biology, 130 1-18. doi:10.1016/bs.mcb.2015.04.007


Author Wall, Adam A.
Condon, Nicholas D.
Yeo, Jeremy C.
Hamilton, Nicholas A.
Stow, Jennifer L.
Title Dynamic imaging of the recycling endosomal network in macrophages
Journal name Methods in Cell Biology   Check publisher's open access policy
ISSN 0091-679X
ISBN 9780128028292
Publication date 2015-06-11
Sub-type Research book chapter (original research)
DOI 10.1016/bs.mcb.2015.04.007
Volume 130
Start page 1
End page 18
Total pages 18
Place of publication Maryland Heights, MO, United States
Publisher Academic Press
Collection year 2016
Language eng
Abstract Recycling endosomes (REs) form an extensive and complex network of subcompartmentalized vesicular and tubular elements that connect with the cell surface and other endosomes in macrophages. As surveillance and defense cells of the innate immune system, macrophages are highly dependent on REs for their active and voluminous cell surface turnover and endocytic, exocytic, and recycling of membrane and cargo. Here we set out three approaches for imaging and analyzing REs in macrophages, based on the expression of fluorescently labeled RE-associated proteins and the uptake of fluorescent cargo. Subcompartments of the REs are identified by co-expression and co-localization analysis of RE associated Rab GTPases. Transferrin is a well-known cargo marker as it recycles through REs and it is compared here to other cargo, revealing how different endocytic routes intersect with REs. We show how the movement of transferrin through REs can be modeled and quantified in live cells. Finally, since phagosomes are a signature organelle for macrophages, and REs fuse with the maturing phagosome, we show imaging of REs with phagosomes using a genetically encoded pH-sensitive SNARE-based probe. Together these approaches provide multiple ways to comprehensively analyze REs and the important roles they play in these immune cells and more broadly in other cell types.
Keyword Endocytosis
Live cell imaging
Macrophage
Phagocytosis
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Article in press corrected proof.

Document type: Journal Article
Sub-type: Research book chapter (original research)
Collections: Official 2016 Collection
Institute for Molecular Bioscience - Publications
 
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