Use of dansyl-cholestanol as a probe of cholesterol behavior in membranes of living cells. Academic Article uri icon

abstract

  • While plasma membrane cholesterol-rich microdomains play a role in cholesterol trafficking, little is known about the appearance and dynamics of cholesterol through these domains in living cells. The fluorescent cholesterol analog 6-dansyl-cholestanol (DChol), its biochemical fractionation, and confocal imaging of L-cell fibroblasts contributed the following new insights: i) fluorescence properties of DChol were sensitive to microenvironment polarity and mobility; (ii) DChol taken up by L-cell fibroblasts was distributed similarly as cholesterol and preferentially into cholesterol-rich vs. -poor microdomains resolved by affinity chromatography of purified plasma membranes; iii) DChol reported similar polarity (dielectric constant near 18) but higher mobility near phospholipid polar head group region for cholesterol in purified cholesterol-rich versus -poor microdomains; and iv) real-time confocal imaging, quantitative colocalization analysis, and fluorescence resonance energy transfer with cholesterol-rich and -poor microdomain markers confirmed that DChol preferentially localized in plasma membrane cholesterol-rich microdomains of living cells. Thus, DChol sensed a unique, relatively more mobile microenvironment for cholesterol in plasma membrane cholesterol-rich microdomains, consistent with the known, more rapid exchange dynamics of cholesterol from cholesterol-rich than -poor microdomains.

published proceedings

  • J Lipid Res

author list (cited authors)

  • Huang, H., McIntosh, A. L., Atshaves, B. P., Ohno-Iwashita, Y., Kier, A. B., & Schroeder, F.

citation count

  • 19

complete list of authors

  • Huang, Huan||McIntosh, Avery L||Atshaves, Barbara P||Ohno-Iwashita, Yoshiko||Kier, Ann B||Schroeder, Friedhelm

publication date

  • May 2010