Monoclonal antibody detection of plasma membrane cholesterol microdomains responsive to cholesterol trafficking

Citation
Hs. Kruth et al., Monoclonal antibody detection of plasma membrane cholesterol microdomains responsive to cholesterol trafficking, J LIPID RES, 42(9), 2001, pp. 1492-1500
Citations number
60
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biochemistry & Biophysics
Journal title
JOURNAL OF LIPID RESEARCH
ISSN journal
0022-2275 → ACNP
Volume
42
Issue
9
Year of publication
2001
Pages
1492 - 1500
Database
ISI
SICI code
0022-2275(200109)42:9<1492:MADOPM>2.0.ZU;2-4
Abstract
The hypothesis of lipid domains in cellular plasma membranes is well establ ished. However, direct visualization of the domains has been difficult. Her e we report direct visualization of plasma membrane cholesterol microdomain s modulated by agents that affect cholesterol trafficking to and from the p lasma membrane. The cholesterol microdomains were visualized with a monoclo nal antibody that specifically detects ordered cholesterol arrays. These un ique cholesterol microdomains were induced on macrophages and fibroblasts w hen they were enriched with cholesterol in the presence of an ACAT inhibito r, to block esterification of excess cellular cholesterol. Induction of the plasma membrane cholesterol microdomains could be blocked by agents that i nhibit trafficking of cholesterol to the plasma membrane and by cholesterol acceptors that remove cholesterol from the plasma membrane. In addition, p lasma membrane cholesterol microdomains did not develop in mutant Niemann-P ick type C fibroblasts, consistent with the defect in cholesterol trafficki ng reported for these cells. The induction of plasma membrane cholesterol m icrodomains on inhibition of ACAT helps explain how AC-AT inhibition promot es cholesterol efflux from cells in the presence of cholesterol acceptors s uch as HDL. The anti-cholesterol monoclonal antibody also detected extracel lular cholesterol-containing particles that accumulated most prominently du ring cholesterol enrichment of less differentiated human monocyte-macrophag es. For the first time, cholesterol microdomains have been visualized that function in cholesterol trafficking to and from the plasma membrane.