Examining glucose transport in single vascular smooth muscle cells with a fluorescent glucose analog.
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.
Lloyd
P G
PG
Department of Physiology and Dalton Cardiovascular Research Center, University of Missouri-Columbia, 65212, USA.
Hardin
C D
CD
Sturek
M
M
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D000818:Animals; D002339:Carotid Arteries / Q000166:cytology; D002467:Cell Nucleus / Q000378:metabolism; D003847:Deoxyglucose / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D005456:Fluorescent Dyes / Q000493:pharmacokinetics*; D005947:Glucose / Q000493:pharmacokinetics*; D007091:Image Processing, Computer-Assisted / Q000379:methods; D019344:Lactic Acid / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism*; D013552:Swine
D005456:Fluorescent Dyes; D019344:Lactic Acid; D003847:Deoxyglucose; D009327:4-Chloro-7-nitrobenzofurazan; D005947:Glucose; C098340:2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose