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Cholesterol Distribution in Living Cells: Fluorescence Imaging Using Dehydroergosterol as a Fluorescent Cholesterol Analog

Mukherjee, Sushmita, Zha, Xiaohui, Tabas, Ira, Maxfield, Frederick R.·1998·Biophysical Journal
DOI 10.1016/s0006-3495(98)77632-5
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Abstract

Cholesterol is an important constituent of most mammalian cell membranes and its concentration in various cellular membranes is tightly regulated. Although there is much information about cholesterol distribution and trafficking in cells, it is primarily derived from indirect measurements, and the results obtained using different approaches are often conflicting. A cholesterol analog that faithfully mimics the properties of cholesterol and can be followed in living cells would thus be very useful. In this study, we report the fluorescence imaging of such an analog, dehydroergosterol (DHE), in living cells. DHE differs from cholesterol in having three additional double bonds and an extra methyl group. In model systems, DHE closely mimics the behavior of native cholesterol. Using triple-labeling studies, we show that DHE colocalizes extensively with endocytosed transferrin, an endocytic recycling compartment marker, and with a marker for the trans-Golgi network, Tac-TGN38. This distribution of DHE is qualitatively similar to that observed when cells are labeled with the fluorescent cholesterol-binding polyene antibiotic, filipin, although there are differences in apparent proportions of DHE and filipin that are localized at the plasma membrane. Another cholesterol derivative, 25-NBD-cholesterol, has a structure that is compromised by the presence of a bulky NBD group and does not distribute to the same organelles as DHE or filipin. In addition, we show in this manuscript that k…

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  • Rab11 participates in the transport of cholesterol through the ERC.

    “Rab11 participates in the transport of cholesterol through this organelle [12]”

    From EHD1 regulates cholesterol homeostasis and lipid droplet storage (2007) · cites this paper as [12]

  • Overexpression of Rab11 causes accumulation of cholesterol in the ERC and inhibits cholesterol esterification.

    “overexpression of Rab11, a key GTPase involved in the regulation of recycling from the ERC, caused the accumulation of cholesterol in this compartment and inhibited cholesterol esterification [12].”

    From EHD1 regulates cholesterol homeostasis and lipid droplet storage (2007) · cites this paper as [12]

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Metadata

Authors
Mukherjee, S., Zha, X., Tabas, I., & Maxfield, F. R.
Venue
Biophysical Journal
Publisher
Elsevier BV
Published
1998-10
Volume
75
Issue
4
Pages
1915-1925
ISSN
0006-3495
DOI
10.1016/s0006-3495(98)77632-5
Cited by
300
References
2
Language
en
Licence

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