Use of BODIPY‐cholesterol (TF‐Chol) for visualizing lysosomal cholesterol accumulation

Abstract
BODIPY‐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 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 LDL particles. When administered to cells from solvent, lysosomal sterol sequestration becomes evident after an overnight equilibration between membranes.

BODIPY‐cholesterol (TopFluor‐Cholesterol) is a widely used cholesterol analogue due to its excellent fluorescence properties and considerable similarity with natural cholesterol in terms of membrane partitioning. However, specific concerns were recently raised about its suitability to detect lysosomal cholesterol accumulation. Here, we provide an explanation to these apparent discrepancies and offer practical suggestions for using this probe in visualizing lysosomal cholesterol accumulation.

Source: Traffic - May 18, 2016 Category: Research Authors: Maarit Hölttä‐Vuori, Erdinc Sezgin, Christian Eggeling, Elina Ikonen Tags: Traffic Interchange Source Type: research