Use of BODIPY-Cholesterol (TF-Chol) for Visualizing Lysosomal Cholesterol Accumulation

Traffic. 2016 Sep;17(9):1054-7. doi: 10.1111/tra.12414. Epub 2016 Jun 13.

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.

Keywords: BODIPY-cholesterol; Niemann-Pick type C disease; TF-cholesterol; cholesterol accumulation; lipid imaging.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Transport
  • Boron Compounds / chemistry*
  • Carrier Proteins / genetics
  • Cell Line
  • Cell Membrane / metabolism*
  • Cholesterol / chemistry
  • Cholesterol / metabolism*
  • Esterification
  • Fibroblasts / metabolism*
  • Humans
  • Intracellular Membranes / metabolism*
  • Lysosomal Storage Diseases / diagnosis
  • Lysosomal Storage Diseases / metabolism
  • Lysosomes / metabolism*
  • Membrane Glycoproteins / genetics
  • Microscopy, Fluorescence

Substances

  • 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene
  • Boron Compounds
  • Carrier Proteins
  • Membrane Glycoproteins
  • NPC1 protein, human
  • Cholesterol