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Sterolight as imaging tool to study sterol uptake, trafficking and efflux in living cells
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SYSNO ASEP 0557101 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Sterolight as imaging tool to study sterol uptake, trafficking and efflux in living cells Author(s) Králová, Jarmila (UMG-J)
Popr, Martin (UMG-J)
Valečka, Jan (UMG-J)
Bartůněk, Petr (UMG-J) RIDNumber of authors 4 Article number 6264 Source Title Scientific Reports. - : Nature Publishing Group - ISSN 2045-2322
Roč. 12, č. 1 (2022)Number of pages 17 s. Publication form Online - E Language eng - English Country GB - United Kingdom Keywords endoplasmic-reticulum ; cellular cholesterol ; plasma-membrane ; intracellular-transport ; lipid rafts ; endocytosis ; caveolin ; dehydroergosterol ; internalization ; homeostasis Subject RIV EB - Genetics ; Molecular Biology OECD category Cell biology R&D Projects EF18_046/0016045 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) LM2018129 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) GA17-02836S GA ČR - Czech Science Foundation (CSF) LM2018130 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support UMG-J - RVO:68378050 UT WOS 000782844000050 DOI 10.1038/s41598-022-10134-x Annotation Information about cholesterol subcellular localization and transport pathways inside cells is essential for understanding and treatment of cholesterol-related diseases. However, there is a lack of reliable tools to monitor it. This work follows the fate of Sterolight, a BODIPY-labelled sterol, within the cell and demonstrates it as a suitable probe for visualization of sterol/lipid trafficking. Sterolight enters cells through an energy-independent process and knockdown experiments suggest caveolin-1 as its potential cellular carrier. Intracellular transport of Sterolight is a rapid process, and transfer from ER and mitochondria to lysosomes and later to lipid droplets requires the participation of active microtubules, as it can be inhibited by the microtubule disruptor nocodazole. Excess of the probe is actively exported from cells, in addition to being stored in lipid droplets, to re-establish the sterol balance. Efflux occurs through a mechanism requiring energy and may be selectively poisoned with verapamil or blocked in cells with mutated cholesterol transporter NPC1. Sterolight is efficiently transferred within and between different cell populations, making it suitable for monitoring numerous aspects of sterol biology, including the live tracking and visualization of intracellular and intercellular transport. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2023 Electronic address https://www.nature.com/articles/s41598-022-10134-x
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