Transmembrane distribution of sterol in the human erythrocyte

Schroeder, Friedhelm; Nemecz, Gyorgy; Wood, W. Gibson; Joiner, Clinton H.; Morrot, Gil; Ayraut-Jarrier, Marise; Devaux, Philippe F.

doi:10.1016/0005-2736(91)90185-b

Cited by 133 publications

(102 citation statements)

References 48 publications

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“…The transbilayer migration rate of DHE in model membranes and plasma membranes is relatively fast, t 1/2 of sec-min depending on the membrane examined (rev. in (12,108,144,145). Rapid transbilayer migration of DHE is also supported by the finding that uptake of DHE into and non-vesicular transport across cultured hepatocyte-derived cells (i.e.…”

Section: Dhe Architecture In Purified Biological Membranesmentioning

confidence: 76%

Section: Dhe Architecture In Purified Biological Membranesmentioning

confidence: 99%

“…in (12,144,145). DHE revealed that the cytofacial leaflet of plasma membranes from L-cell fibroblasts (12,24,146,147), erythrocytes (144), and brain synaptosomes (145,(148)(149)(150)(151). While other methods (filipin staining, exchange, cholesterol oxidase accessibility, neutron diffraction, etc) have also been used to measure transbilayer cholesterol distribution, many have significant limitations including accessibility of membrane cholesterol (filipin, cholesterol oxidase), production of a potential perturbant (cholesterol oxidase produces cholesterone), and poor dynamic resolution (neutron diffraction) (rev.…”

Section: Dhe Architecture In Purified Biological Membranesmentioning

confidence: 99%

“…in (12). While measurements of transbilayer sterol distribution by different methods may therefore not necessarily agree, multiple methods (DHE, NBD-cholesterol quenching, and exchange assays) indicate that the cytofacial leaflet of erythrocytes has more cholesterol than the exofacial leaflet (144,152). Likewise, several methods (DHE, cholesterol oxidase) indicate that the synaptosomal plasma membrane has more cholesterol in the cytofacial leaflet than the exofacial leaflet (12,145,(148)(149)(150).…”

Section: Dhe Architecture In Purified Biological Membranesmentioning

confidence: 99%

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Fluorescence Techniques Using Dehydroergosterol to Study Cholesterol Trafficking

McIntosh

Atshaves

Huang

et al. 2008

Lipids

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Cholesterol itself has very few structural/chemical features suitable for real-time imaging in living cells. Thus, the advent of dehydroergosterol [ergosta-5,7,9(11),22-tetraen-3β-ol, DHE] the fluorescent sterol most structurally and functionally similar to cholesterol to date, has proven to be a major asset for real-time probing/elucidating the sterol environment and intracellular sterol trafficking in living organisms. DHE is a naturally-occurring, fluorescent sterol analog that faithfully mimics many of the properties of cholesterol. Because these properties are very sensitive to sterol structure and degradation, such studies require the use of extremely pure (>98%) quantities of fluorescent sterol. DHE is readily bound by cholesterol-binding proteins, is incorporated into lipoproteins (from the diet of animals or by exchange in vitro), and for real-time imaging studies is easily incorporated into cultured cells where it co-distributes with endogenous sterol. Incorporation from an ethanolic stock solution to cell culture media is effective, but this process forms an aqueous dispersion of dehydroergosterol crystals which can result in endocytic cellular uptake and distribution into lysosomes which is problematic in imaging DHE at the plasma membrane of living cells. In contrast, monomeric DHE can be incorporated from unilamellar vesicles by exchange/fusion with the plasma membrane or from DHE-methyl-β-cyclodextrin (DHE-MβCD) complexes by exchange with the plasma membrane. Both of the latter techniques can deliver large quantities of monomeric dehydroergosterol with significant distribution into the plasma membrane. The properties and behavior of DHE in protein-binding, lipoproteins, model membranes, biological membranes, lipid rafts/caveolae, and real-time imaging in living cells indicate that this naturally-occurring fluorescent sterol is a useful mimic for probing the properties of cholesterol in these systems. Keywordsdehydroergosterol; cholesterol; ergosterol; fluorescent sterols; microscopy Historical PerspectiveIn order to resolve the dynamics and factors governing cholesterol trafficking within cells, it is important to recognize that the cholesterol molecule has very few polar constituents (Fig. 1A). Consequently, cholesterol is poorly soluble in aqueous environments such as cytosol as evidenced by critical micellar concentrations of cholesterol and other sterols being very low, *To whom correspondence should be addressed: Department of Physiology and Pharmacology, Texas A&M University, TVMC, College Station, TX 862-1433, FAX: (979) NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript near 20-30 nM (1-5). The physiological impact of cholesterol's low aqueous solubility is that spontaneous cholesterol desorption from the cytofacial leaflet of the plasma membrane or lysosomal membrane into the cytosol for transfer to intracellular sites for esterification, oxidation, or secretion is extremely slow (t 1/2 3 hours-days) (6-9). Therefore, it is important to resolve factors...

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Section: Dhe Architecture In Purified Biological Membranesmentioning

confidence: 76%

Section: Dhe Architecture In Purified Biological Membranesmentioning

confidence: 99%

Section: Dhe Architecture In Purified Biological Membranesmentioning

confidence: 99%

Section: Dhe Architecture In Purified Biological Membranesmentioning

confidence: 99%

See 2 more Smart Citations

Fluorescence Techniques Using Dehydroergosterol to Study Cholesterol Trafficking

McIntosh

Atshaves

Huang

et al. 2008

Lipids

Self Cite

View full text Add to dashboard Cite

show abstract

“…Besides phospholipids, plasma membranes contain a high amount of cholesterol, but the distribution of this lipid is still a subject of debate: some reports are in favour of an almost symmetrical repartition in the two leaflets (Blau and Pelletier et al, 1987Post et al, 1988Record et al, 1984Higgins and Evans, 1978Sessions and Horwitz, 1983Sessions and Horwitz, 1983Sessions and Horwitz, 1983 Olaisson et al, 1985Deutsch and Kelly, 1981Bollen and Higgins, 1980Dominski et al, 1983Buckland et al, 1978Marinetti et al, 1976Nilsson and Dallner, 1977 Crain and Marinetti, 1979Krebs et al, 1979Harb et al, 1981 an uneven distribution of cholesterol (Brasaemle et al, 1988;Fisher, 1976;Schroeder et al, 1991).…”

Section: Asymmetry In Various Membranes Plasma Membranesmentioning

confidence: 99%