Kinetics of contraction-induced GLUT4 translocation in skeletal muscle fibers from living mice.
OBJECTIVE
Exercise is an important strategy for the treatment of type 2 diabetes. This is due in part to an increase in glucose transport that occurs in the working skeletal muscles. Glucose transport is regulated by GLUT4 translocation in muscle, but the molecular machinery mediating this process is poorly understood. The purpose of this study was to 1) use a novel imaging system to elucidate the kinetics of contraction-induced GLUT4 translocation in skeletal muscle and 2) determine the function of AMP-activated protein kinase alpha2 (AMPKalpha2) in this process.


METHODS
Confocal imaging was used to visualize GLUT4-enhanced green fluorescent protein (EGFP) in transfected quadriceps muscle fibers in living mice subjected to contractions or the AMPK-activator AICAR.


RESULTS
Contraction increased GLUT4-EGFP translocation from intracellular vesicle depots to both the sarcolemma and t-tubules with similar kinetics, although translocation was greater with contractions elicited by higher voltage. Re-internalization of GLUT4 did not begin until 10 min after contractions ceased and was not complete until 130 min after contractions. AICAR increased GLUT4-EGFP translocation to both sarcolemma and t-tubules with similar kinetics. Ablation of AMPKalpha2 activity in AMPKalpha2 inactive transgenic mice did not change GLUT4-EGFP's basal localization, contraction-stimulated intracellular GLUT4-EGFP vesicle depletion, translocation, or re-internalization, but diminished AICAR-induced translocation.


CONCLUSIONS
We have developed a novel imaging system to study contraction-stimulated GLUT4 translocation in living mice. Contractions increase GLUT4 translocation to the sarcolemma and t-tubules with similar kinetics and do not require AMPKalpha2 activity.
Lauritzen
Hans P M M
HP
Research Division, Joslin Diabetes Center and Harvard Medical School, Boston, Massachusetts, USA.
Galbo
Henrik
H
Toyoda
Taro
T
Goodyear
Laurie J
LJ
D000263:Adenylate Kinase / Q000378:metabolism; D000818:Animals; D051275:Glucose Transporter Type 4 / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007700:Kinetics; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008870:Microtubules / Q000378:metabolism; D018485:Muscle Fibers, Skeletal / Q000502:physiology*; D010957:Plasmids; D021381:Protein Transport / Q000502:physiology; D012508:Sarcolemma / Q000378:metabolism; D014162:Transfection
D051275:Glucose Transporter Type 4; D049452:Green Fluorescent Proteins; D000263:Adenylate Kinase