title: A Molecular Probe for the Detection of Polar Lipids in Live Cells.
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.
authors.affiliation: School of Pharmacy and Medical Science, University of South Australia, Adelaide, South Australia, Australia.
authors.forename: Christie A
authors.initials: CA
authors.lastname: Bader
authors.affiliation: School of Pharmacy and Medical Science, University of South Australia, Adelaide, South Australia, Australia.
authors.forename: Tetyana
authors.initials: T
authors.lastname: Shandala
authors.affiliation: Vibrational Spectroscopy Core Facility, The University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Carter
authors.affiliation: Future Industries Institute, University of South Australia, Mawson Lakes, South Australia, Australia.
authors.forename: Angela
authors.initials: A
authors.lastname: Ivask
authors.affiliation: Future Industries Institute, University of South Australia, Mawson Lakes, South Australia, Australia.
authors.forename: Taryn
authors.initials: T
authors.lastname: Guinan
authors.affiliation: School of Pharmacy and Medical Science, University of South Australia, Adelaide, South Australia, Australia.
authors.forename: Shane M
authors.initials: SM
authors.lastname: Hickey
authors.affiliation: Department of Chemistry and Nanochemistry Research Institute, Curtin University, Bentley, Western Australia, Australia.
authors.forename: Melissa V
authors.initials: MV
authors.lastname: Werrett
authors.affiliation: Department of Chemistry and Nanochemistry Research Institute, Curtin University, Bentley, Western Australia, Australia.
authors.forename: Phillip J
authors.initials: PJ
authors.lastname: Wright
authors.affiliation: Department of Chemistry and Nanochemistry Research Institute, Curtin University, Bentley, Western Australia, Australia.
authors.forename: Peter V
authors.initials: PV
authors.lastname: Simpson
authors.affiliation: Department of Industrial Chemistry "Toso Montanari", University of Bologna, Bologna, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Stagni
authors.affiliation: Future Industries Institute, University of South Australia, Mawson Lakes, South Australia, Australia.
authors.forename: Nicolas H
authors.initials: NH
authors.lastname: Voelcker
authors.affiliation: Vibrational Spectroscopy Core Facility, The University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Peter A
authors.initials: PA
authors.lastname: Lay
authors.affiliation: Department of Chemistry and Nanochemistry Research Institute, Curtin University, Bentley, Western Australia, Australia.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Massi
authors.affiliation: School of Pharmacy and Medical Science, University of South Australia, Adelaide, South Australia, Australia.
authors.forename: Sally E
authors.identifier: http://orcid.org/0000-0002-9999-9154
authors.initials: SE
authors.lastname: Plush
authors.affiliation: School of Pharmacy and Medical Science, University of South Australia, Adelaide, South Australia, Australia.
authors.forename: Douglas A
authors.initials: DA
authors.lastname: Brooks
mesh_terms: D017667:Adipocytes; D000273:Adipose Tissue / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D001343:Autophagy; D001692:Biological Transport; D050260:Carbohydrate Metabolism; D002468:Cell Physiological Phenomena; D004330:Drosophila; D066292:Lipid Droplets / Q000378:metabolism; D050356:Lipid Metabolism; D008055:Lipids / Q000737:chemistry; D008675:Metamorphosis, Biological; D051379:Mice; D015335:Molecular Probes; D013059:Spectrum Analysis, Raman; D013194:Staining and Labeling
chemical_list: D000596:Amino Acids; D008055:Lipids; D015335:Molecular Probes