Measuring Both Glucose Uptake and Myosin Heavy Chain Isoform Expression in Single Rat Skeletal Muscle Fibers
Glucose uptake by skeletal muscle is important for metabolic health. Because skeletal muscle is composed of multiple fiber types that have differing metabolic and contractile properties, studying glucose uptake in whole muscle tissue does not elucidate differences at the cellular level. Here, we describe a procedure that enables the measurement of both glucose uptake and fiber type (by myosin heavy chain isoform expression) in individual rat epitrochlearis muscle fibers.
Key wordsMuscle fiber type Myosin heavy chain Glucose transport Glucose uptake SDS-PAGE
This work was supported by grants from the National Institutes of Health (R01 DK71771 and R01 AG10026). We thank Jim MacKrell and Haiyan Wang for providing valuable suggestions about the manuscript.
- 11.Funai K, Cartee GD (2009) Inhibition of contraction-stimulated AMP-activated protein kinase inhibits contraction-stimulated increases in PAS-TBC1D1 and glucose transport without altering PAS-AS160 in rat skeletal muscle. Diabetes 58(5):1096–1104. https://doi.org/10.2337/db08-1477CrossRefPubMedPubMedCentralGoogle Scholar
- 16.Wang H, Arias EB, Yu CS, Verkerke AR, Cartee GD (2017) Effects of calorie restriction and fiber type on glucose uptake and abundance of electron transport chain and oxidative phosphorylation proteins in single fibers from old rats. J Gerontol Ser A: Biomed Sci Med Sci 72(12):1638–1646CrossRefGoogle Scholar
- 22.Castorena CM, Mackrell JG, Bogan JS, Kanzaki M, Cartee GD (2011) Clustering of GLUT4, TUG, and RUVBL2 protein levels correlate with myosin heavy chain isoform pattern in skeletal muscles, but AS160 and TBC1D1 levels do not. J Appl Physiol (1985) 111(4):1106–1117. https://doi.org/10.1152/japplphysiol.00631.2011CrossRefPubMedCentralGoogle Scholar