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Use of dansyl-cholestanol as a probe of cholesterol behavior in membranes of living cells.
- Source :
-
Journal of lipid research [J Lipid Res] 2010 May; Vol. 51 (5), pp. 1157-72. Date of Electronic Publication: 2009 Dec 11. - Publication Year :
- 2010
-
Abstract
- While plasma membrane cholesterol-rich microdomains play a role in cholesterol trafficking, little is known about the appearance and dynamics of cholesterol through these domains in living cells. The fluorescent cholesterol analog 6-dansyl-cholestanol (DChol), its biochemical fractionation, and confocal imaging of L-cell fibroblasts contributed the following new insights: i) fluorescence properties of DChol were sensitive to microenvironment polarity and mobility; (ii) DChol taken up by L-cell fibroblasts was distributed similarly as cholesterol and preferentially into cholesterol-rich vs. -poor microdomains resolved by affinity chromatography of purified plasma membranes; iii) DChol reported similar polarity (dielectric constant near 18) but higher mobility near phospholipid polar head group region for cholesterol in purified cholesterol-rich versus -poor microdomains; and iv) real-time confocal imaging, quantitative colocalization analysis, and fluorescence resonance energy transfer with cholesterol-rich and -poor microdomain markers confirmed that DChol preferentially localized in plasma membrane cholesterol-rich microdomains of living cells. Thus, DChol sensed a unique, relatively more mobile microenvironment for cholesterol in plasma membrane cholesterol-rich microdomains, consistent with the known, more rapid exchange dynamics of cholesterol from cholesterol-rich than -poor microdomains.
- Subjects :
- Animals
Biological Transport
Biomarkers metabolism
Buffers
Cell Survival
Chromatography, Affinity
Fluorescence Resonance Energy Transfer
L Cells
Membrane Microdomains metabolism
Mice
Sterols chemistry
Sterols metabolism
Time Factors
Unilamellar Liposomes metabolism
Water metabolism
Cell Membrane metabolism
Cholestanols metabolism
Cholesterol metabolism
Fluorescent Dyes metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1539-7262
- Volume :
- 51
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Journal of lipid research
- Publication Type :
- Academic Journal
- Accession number :
- 20008119
- Full Text :
- https://doi.org/10.1194/jlr.M003244