Examining glucose transport in single vascular smooth muscle cells with a fluorescent glucose analog

Physiol Res. 1999;48(6):401-10.

Abstract

Changes in vascular smooth muscle glucose transport are thought to contribute to the pathogenesis of vascular disease in conditions such as diabetes, yet no single-cell assay for glucose uptake by VSM exists. Therefore, we examined the uptake of the fluorescent glucose analog 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxyglucose (2-NBDG) in isolated pig vascular smooth muscle cells (VSMC) using digital imaging microscopy. Uptake of 2-NBDG by VSMC was inhibited by D-glucose but not by L-glucose, suggesting that 2-NBDG enters VSMC via glucose transporters. Uptake of 2-NBDG was linear in the presence of 10 mM D-glucose (n=6, R2=0.9408) but not in its absence (n=4, R2=0.9993), indicating that 2-NBDG is not metabolized and accumulates within the cells. 2-NBDG fluorescence in VSMC was often non-uniform and appeared to represent binding of 2-NBDG to some cytoplasmic component. The present study demonstrates that 2-NBDG is a useful tool for examining vascular smooth muscle glucose uptake at the single cell level.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 4-Chloro-7-nitrobenzofurazan / analogs & derivatives*
  • 4-Chloro-7-nitrobenzofurazan / pharmacokinetics
  • Animals
  • Carotid Arteries / cytology
  • Cell Nucleus / metabolism
  • Deoxyglucose / analogs & derivatives*
  • Deoxyglucose / pharmacokinetics
  • Fluorescent Dyes / pharmacokinetics*
  • Glucose / pharmacokinetics*
  • Image Processing, Computer-Assisted / methods
  • Lactic Acid / metabolism
  • Microscopy, Fluorescence / methods
  • Muscle, Smooth, Vascular / cytology
  • Muscle, Smooth, Vascular / metabolism*
  • Swine

Substances

  • Fluorescent Dyes
  • Lactic Acid
  • Deoxyglucose
  • 4-Chloro-7-nitrobenzofurazan
  • Glucose
  • 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose