J. Lipid Res. Neurobiology of Lipids (ISSN1683-5506)
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Originally published In Press as doi:10.1194/jlr.M300033-JLR200 on March 16, 2003

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Journal of Lipid Research, Vol. 44, 1094-1099, June 2003
Copyright © 2003 by Lipid Research, Inc.

Visualizing caveolin-1 and HDL in cholesterol-loaded aortic endothelial cells

W. T. Chao*, S. S. Fan*, J. K. Chen{dagger} and V. C. Yang1,*

* Department of Biology and Life Science Research Center, Tunghai University, Taichung, Taiwan, Republic of China
{dagger} Department of Physiology, College of Medicine, Chang Gung University, Taoyuan, Taiwan, Republic of China

1 To whom correspondence should be addressed. e-mail: vcyang{at}mail.thu.edu.tw

Caveolae are vesicular invaginations of the plasma membranes that regulate signal transduction and transcytosis, as well as cellular cholesterol homeostasis. Our previous studies indicated that the removal of cholesterol from aortic endothelial cells and smooth muscle cells in the presence of HDL is associated with plasmalemmal invaginations and plasmalemmal vesicles. The goal of the present study was to investigate the location and distribution of caveolin-1, the main structural protein component of caveolae, in cholesterol-loaded aortic endothelial cells after HDL incubation. Confocal microscopic analysis demonstrated that the caveolin-1 appeared to colocalize with HDL-fluorescein 1,1'-dioctadecyl 3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) conjugates on the cell surface. No free HDL-DiI conjugates were revealed in the cytoplasm. Immunoelectron microscopy further demonstrated that caveolin-1 gold (15 nm) conjugates colocalized with HDL gold (10 nm) conjugates in the plasmalemmal invaginations.

These morphological results indicated that caveolae are the major membrane domains facilitating the transport of excess cholesterol to HDL on the cell surface of aortic endothelial cells.

Supplementary key words high density lipoprotein • colocalization • atherosclerosis


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