Membrane organization and intracellular transport of a fluorescent analogue of 27-hydroxycholesterol

Szomek, Maria; Moesgaard, Laust; Reinholdt, Peter; Hald, Sophia Bell Haarhøj; Petersen, Daniel C.; Krishnan, Kathiresan; Covey, Douglas F.; Kongsted, Jacob; Wüstner, Daniel

doi:10.1016/j.chemphyslip.2020.105004

Cited by 9 publications

(13 citation statements)

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“…This enabled us to study the effect of one additional hydroxy group in the steroid side chain on intracellular sterol trafficking and efflux. We found that 27-OH-CTL and CTL show a similar extent of non-vesicular transport, as measured by fluorescence recovery after photobleaching (not shown but see Szomek et al (2020) ). However, 27-OH-CTL was much more efficiently effluxed from cells than CTL ( Figure 5A ) and 27-OH-CTL but not CTL accumulated a lot in LDs in human fibroblasts ( Szomek et al, 2020 ).…”

Section: Formation and Efflux Of Oxysterolssupporting

confidence: 53%

“…We found that 27-OH-CTL and CTL show a similar extent of non-vesicular transport, as measured by fluorescence recovery after photobleaching (not shown but see Szomek et al (2020) ). However, 27-OH-CTL was much more efficiently effluxed from cells than CTL ( Figure 5A ) and 27-OH-CTL but not CTL accumulated a lot in LDs in human fibroblasts ( Szomek et al, 2020 ). Comparable intracellular transport kinetics but more rapid and extensive efflux, as well as the metabolic conversion, was also found for 25-hydroxycholesterol compared to cholesterol in macrophages using radioactive sterol probes ( Morel et al, 1996 ).…”

Section: Formation and Efflux Of Oxysterolssupporting

confidence: 53%

“…Its export from lysosomes depends on NPC1 and to a lower extent on NPC2 ( Iaea et al, 2015 ; Petersen et al, 2020 ). Based on the same design strategy, we have recently developed a novel derivative of 27-hydroxycholesterol, 27-hydroxycholestatrienol (27-OH-CTL), which differs from 27-hydroxycholesterol only by two additional double bonds in the steroid ring system ( Figure 5 ), allowing for spectroscopic analysis and live-cell imaging by ultraviolet-sensitive microscopy, similar to 25-hydroxycholestatrienol ( Szomek et al, 2020 ). We compared the intracellular trafficking of 27-OH-CTL with that of cholestatrienol (CTL), which has identical photophysical properties and just two additional double bonds compared to cholesterol ( Figure 5 ).…”

Section: Formation and Efflux Of Oxysterolsmentioning

confidence: 99%

“…Extra double bonds in the fluorescent analogs are the only modifications compared to the natural sterols and are shown in blue. Figure adapted and reproduced from ( Szomek et al, 2020 ) with permission.…”

Section: Formation and Efflux Of Oxysterolsmentioning

confidence: 99%

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Pathways and Mechanisms of Cellular Cholesterol Efflux—Insight From Imaging

Juhl

Wüstner

2022

Front. Cell Dev. Biol.

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Cholesterol is an essential molecule in cellular membranes, but too much cholesterol can be toxic. Therefore, mammalian cells have developed complex mechanisms to remove excess cholesterol. In this review article, we discuss what is known about such efflux pathways including a discussion of reverse cholesterol transport and formation of high-density lipoprotein, the function of ABC transporters and other sterol efflux proteins, and we highlight their role in human diseases. Attention is paid to the biophysical principles governing efflux of sterols from cells. We also discuss recent evidence for cholesterol efflux by the release of exosomes, microvesicles, and migrasomes. The role of the endo-lysosomal network, lipophagy, and selected lysosomal transporters, such as Niemann Pick type C proteins in cholesterol export from cells is elucidated. Since oxysterols are important regulators of cellular cholesterol efflux, their formation, trafficking, and secretion are described briefly. In addition to discussing results obtained with traditional biochemical methods, focus is on studies that use established and novel bioimaging approaches to obtain insight into cholesterol efflux pathways, including fluorescence and electron microscopy, atomic force microscopy, X-ray tomography as well as mass spectrometry imaging.

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Section: Formation and Efflux Of Oxysterolssupporting

confidence: 53%

Section: Formation and Efflux Of Oxysterolssupporting

confidence: 53%

Section: Formation and Efflux Of Oxysterolsmentioning

confidence: 99%

Section: Formation and Efflux Of Oxysterolsmentioning

confidence: 99%

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Pathways and Mechanisms of Cellular Cholesterol Efflux—Insight From Imaging

Juhl

Wüstner

2022

Front. Cell Dev. Biol.

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“…This is consistent with previous experimental and computational work that suggests cholesterol has a preferred tilt angle of 20−30°. [40][41][42]44,76 Minor differences are observed in the average tilt angle of both cholesterol and oxidized cholesterol. In the neuronal membrane, cholesterol has tilt angles of 147.86 ± 20.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Site of Cholesterol Oxidation Impacts Its Localization and Domain Formation in the Neuronal Plasma Membrane

Wilson

Wang

O’Mara

2021

ACS Chem. Neurosci.

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Cholesterol is integral to the structure of mammalian cell membranes. Oxidation of cholesterol alters how it behaves in the membrane and influences the membrane biophysical properties. Elevated levels of oxidized cholesterol are associated with neurodegenerative diseases such as Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, and Huntington’s disease. Previous work has investigated the impact of oxidized cholesterol in the context of simple model membrane systems. However, there is a growing body of literature that shows that complex membranes possessing physiological phospholipid distributions have different properties from those of binary or trinary model membranes. In the current work, the impact of oxidized cholesterol on the biophysical properties of a complex neuronal plasma membrane is investigated using coarse-grained Martini molecular dynamics simulations. Comparison of the native neuronal membrane to neuronal membranes containing 10% tail-oxidized or 10% head-oxidized cholesterol shows that the site of oxidization changes the behavior of the oxidized cholesterol in the membrane. Furthermore, species-specific domain formation is observed between each oxidized cholesterol and minor lipid classes. Although both tail-oxidized and head-oxidized cholesterols modulate the biophysical properties of the membrane, smaller changes are observed in the complex neuronal membrane than seen in the previous work on simple binary or trinary model membranes. This work highlights the presence of compensatory effects of lipid diversity in the complex neuronal membrane. Overall, this study improves our molecular-level understanding of the effects of oxidized cholesterol on the properties of neuronal tissue and emphasizes the importance of studying membranes with realistic lipid compositions.

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