Králová Jarmila, Popr Martin, Valečka Jan, Bartůněk Petr
CZ-OPENSCREEN, Institute of Molecular Genetics of the Czech Academy of Sciences, v.v.i., Vídeňská 1083, 142 20, Prague 4, Czech Republic.
Light Microscopy Core Facility, Institute of Molecular Genetics of the Czech Academy of Sciences, v.v.i., Vídeňská 1083, 142 20, Prague 4, Czech Republic.
Sci Rep. 2022 Apr 15;12(1):6264. doi: 10.1038/s41598-022-10134-x.
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.
了解细胞内胆固醇的亚细胞定位和转运途径对于理解和治疗胆固醇相关疾病至关重要。然而，目前缺乏可靠的工具来监测这一过程。本研究追踪了一种BODIPY标记的固醇Sterolight在细胞内的命运，并证明它是一种用于可视化固醇/脂质运输的合适探针。Sterolight通过能量非依赖过程进入细胞，敲低实验表明小窝蛋白-1是其潜在的细胞载体。Sterolight的细胞内运输是一个快速过程，从内质网和线粒体转移到溶酶体，随后再到脂滴需要活跃微管的参与，因为它可被微管破坏剂诺考达唑抑制。除了储存在脂滴中，过量的探针会被主动排出细胞，以重新建立固醇平衡。外排通过一种需要能量的机制发生，并且可能被维拉帕米选择性中毒或在具有突变胆固醇转运蛋白NPC1的细胞中被阻断。Sterolight能在不同细胞群体内部和之间有效转移，使其适用于监测固醇生物学的多个方面，包括细胞内和细胞间运输的实时追踪和可视化。
