title: Use of BODIPY-Cholesterol (TF-Chol) for Visualizing Lysosomal Cholesterol Accumulation.
abstract: Dipyrromethene difluoride-cholesterol (TopFluor-Cholesterol, TF-Chol) is a widely used cholesterol analogue due to its excellent fluorescence properties and considerable similarity with natural cholesterol in terms of membrane partitioning. However, the suitability of TF-Chol for detecting lysosomal cholesterol deposition has recently been questioned. Here, we highlight the fact that the method of lipid delivery and the analysis of time-point both affect the membrane distribution and labeling pattern of TF-Chol, similarly as with radiolabeled cholesterol. Lysosomal sterol accumulation characteristic to a lysosomal storage disease is most readily detected when the probe is introduced via the physiological route, i.e. as a sterol fatty acid ester in low-density lipoprotein particles. When administered to cells from solvent, lysosomal sterol sequestration becomes evident after an overnight equilibration between membranes.
authors.affiliation: Faculty of Medicine, Department of Anatomy, University of Helsinki, 00014 Helsinki, Finland.|Minerva Foundation Institute for Medical Research, 00290 Helsinki, Finland.
authors.forename: Maarit
authors.initials: M
authors.lastname: Hölttä-Vuori
authors.affiliation: MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, OX39DS Oxford, UK.
authors.forename: Erdinc
authors.initials: E
authors.lastname: Sezgin
authors.affiliation: MRC Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, OX39DS Oxford, UK.
authors.forename: Christian
authors.initials: C
authors.lastname: Eggeling
authors.affiliation: Faculty of Medicine, Department of Anatomy, University of Helsinki, 00014 Helsinki, Finland.|Minerva Foundation Institute for Medical Research, 00290 Helsinki, Finland.
authors.forename: Elina
authors.initials: E
authors.lastname: Ikonen
mesh_terms: D001692:Biological Transport; D001896:Boron Compounds / Q000737:chemistry*; D002352:Carrier Proteins / Q000235:genetics; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism*; D004951:Esterification; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins; D016464:Lysosomal Storage Diseases / Q000175:diagnosis / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000235:genetics; D008856:Microscopy, Fluorescence; D000077296:Niemann-Pick C1 Protein
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; C106881:NPC1 protein, human; D000077296:Niemann-Pick C1 Protein; D002784:Cholesterol
keywords: BODIPY-cholesterol; Niemann-Pick type C disease; TF-cholesterol; cholesterol accumulation; lipid imaging