A fluorescent cholesterol analogue for observation of free cholesterol in the plasma membrane of live cells

Free cholesterol in mammalian cells resides mostly in the plasma membrane, where it plays an important role in cellular homeostasis. We synthesized a new fluorescent cholesterol analogue that retained an intact alkyl chain and the sterane backbone of cholesterol. The hydroxyl group of cholesterol wa...

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Bibliographic Details
Published in:Analytical biochemistry 2016-01, Vol.492, p.49-55
Main Authors: Ogawa, Yoshikatsu, Tanaka, Mutsuo
Format: Article
Language:English
Subjects:
Animals
Boron Compounds - chemistry
Cell Membrane - chemistry
Cell Membrane - metabolism
Cell Proliferation - drug effects
Cell Survival - drug effects
Chemical probe
Cholesterol - analogs & derivatives
Cholesterol - metabolism
Cholesterol - toxicity
Detergent-resistant membrane
Dogs
Fluorescent Dyes - chemistry
Live-cell imaging
Madin Darby Canine Kidney Cells
Microscopy
Microscopy, Fluorescence
Sterol
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Summary:Free cholesterol in mammalian cells resides mostly in the plasma membrane, where it plays an important role in cellular homeostasis. We synthesized a new fluorescent cholesterol analogue that retained an intact alkyl chain and the sterane backbone of cholesterol. The hydroxyl group of cholesterol was converted into an amino group that was covalently linked to the fluorophore tetramethylrhodamine to retain the ability to form hydrogen bonds with adjacent molecules. Incubating live MDCK (Madin–Darby canine kidney) cells with our fluorescent cholesterol analogue resulted in the generation of intense signals that were detected by microscopy at the plasma membrane. Incubation with the analogue exerted minimal, if any, influence on cell growth, indicating that it could serve as a useful tool for analyzing free cholesterol at the plasma membrane.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2015.09.003