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Type: Journal article
Title: A molecular probe for the detection of polar lipids in live cells
Author: Bader, C.
Shandala, T.
Carter, E.
Ivask, A.
Guinan, T.
Hickey, S.
Werrett, M.
Wright, P.
Simpson, P.
Stagni, S.
Voelcker, N.
Lay, P.
Massi, M.
Plush, S.
Brooks, D.
Citation: PLoS One, 2016; 11(8):e0161557-1-e0161557-24
Publisher: Public Library of Science (PLoS)
Issue Date: 2016
ISSN: 1932-6203
Editor: Witt, S.N.
Statement of
Christie A. Bader, Tetyana Shandala, Elizabeth A. Carter, Angela Ivask, Taryn Guinan, Shane M. Hickey, Melissa V. Werrett, Phillip J. Wright, Peter V. Simpson, Stefano Stagni, Nicolas H. Voelcker, Peter A. Lay, Massimiliano Massi, Sally E. Plush, Douglas A. Brooks
Abstract: Lipids have an important role in many aspects of cell biology, including membrane architecture/compartment formation, intracellular traffic, signalling, hormone regulation, inflammation, energy storage and metabolism. Lipid biology is therefore integrally involved in major human diseases, including metabolic disorders, neurodegenerative diseases, obesity, heart disease, immune disorders and cancers, which commonly display altered lipid transport and metabolism. However, the investigation of these important cellular processes has been limited by the availability of specific tools to visualise lipids in live cells. Here we describe the potential for ReZolve-L1™ to localise to intracellular compartments containing polar lipids, such as for example sphingomyelin and phosphatidylethanolamine. In live Drosophila fat body tissue from third instar larvae, ReZolve-L1™ interacted mainly with lipid droplets, including the core region of these organelles. The presence of polar lipids in the core of these lipid droplets was confirmed by Raman mapping and while this was consistent with the distribution of ReZolve-L1™ it did not exclude that the molecular probe might be detecting other lipid species. In response to complete starvation conditions, ReZolve-L1™ was detected mainly in Atg8-GFP autophagic compartments, and showed reduced staining in the lipid droplets of fat body cells. The induction of autophagy by Tor inhibition also increased ReZolve-L1™ detection in autophagic compartments, whereas Atg9 knock down impaired autophagosome formation and altered the distribution of ReZolve-L1™. Finally, during Drosophila metamorphosis fat body tissues showed increased ReZolve-L1™ staining in autophagic compartments at two hours post puparium formation, when compared to earlier developmental time points. We concluded that ReZolve-L1™ is a new live cell imaging tool, which can be used as an imaging reagent for the detection of polar lipids in different intracellular compartments.
Keywords: Live Cells
Rights: © 2016 Bader et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
DOI: 10.1371/journal.pone.0161557
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