3-Hexanoyl-7-nitrobenz-2-oxa-1,3-diazol-4-yl-cholesterol (3-NBD-cholesterol) is a versatile cholesterol tracer

Steroids. 2021 Jul:171:108840. doi: 10.1016/j.steroids.2021.108840. Epub 2021 Apr 20.

Abstract

Membrane cholesterol influences a large number of cellular processes and the dynamics of cholesterol exchange between membranes is an area of active study. However, analogs containing a fluorophore on the isooctyl side chain of cholesterol are commonly used without regard for the potential impact of the fluorophore on membrane structure. We investigated the capacity of 3-hexanoyl-7-nitrobenz-2-oxa-1,3-diazol-4-yl-cholesterol (3-NBD-cholesterol), which is labelled at the C3 position, to trace cholesterol dynamics in cellular systems. Transfer of 3-NBD-cholesterol from erythrocytes to lipoproteins replicated known properties of cholesterol. Labelled cells were also readily detected by flow-cytometry and microscopy. Using flow-cytometry it was also possible to follow the uptake of 3-NBD-cholesterol labelled extracellular vesicles. These data indicate that 3-NBD-cholesterol is a versatile cholesterol tracer in different cell models and extracellular vesicles.

Keywords: Cholesterol; Exosome; Extracellular vesicles; Fluorescent cholesterol; Microparticles; NBD.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 4-Chloro-7-nitrobenzofurazan / analogs & derivatives*
  • Azoles
  • Cholesterol / analogs & derivatives*
  • Fluorescent Dyes
  • Nitrobenzenes

Substances

  • 7-nitrobenz-2-oxa-1,3-diazol-4-yl
  • 7-nitrobenz-2-oxa-1,3-diazol-4-ylcholesterol
  • Azoles
  • Fluorescent Dyes
  • Nitrobenzenes
  • Cholesterol
  • 4-Chloro-7-nitrobenzofurazan