Douen A G, Ramlal T, Rastogi S, Bilan P J, Cartee G D, Vranic M, Holloszy J O, Klip A
Division of Cell Biology, Hospital for Sick Children, Toronto, Ontario, Canada.
J Biol Chem. 1990 Aug 15;265(23):13427-30.
Acute exercise, like insulin, increases D-glucose uptake into rat hind limb muscles. Here we examine the distribution of the muscle glucose transporters GLUT-4 and GLUT-1 in plasma membrane and intracellular membrane fractions of skeletal muscle prepared from control, exercised, and acutely insulin-treated rats. Immunoblotting with an anti-GLUT-4 polyclonal antibody showed that acute insulin treatment (by hind limb perfusion or in vivo injection) increased GLUT-4 transporters in a plasma membrane fraction and decreased them in an intracellular membrane fraction. Exercise also increased the GLUT-4 transporters in the plasma membrane, but in contrast to insulin, did not significantly decrease them in the intracellular fraction. Immunoblotting with anti-GLUT-1 antibody revealed that this transporter is largely localized in the plasma membrane. Neither insulin nor exercise significantly increased GLUT-1 transporters in the plasma membrane. The data show that GLUT-4 is an insulin-responsive glucose transporter in skeletal muscle and, furthermore, that GLUT-4 also responds to acute exercise. The results are consistent with recruitment of GLUT-4 glucose transporters to the plasma membrane from intracellular stores. Moreover, exercise-sensitive GLUT-4 transporters do not originate from the insulin-sensitive intracellular membrane fraction, suggesting the existence of distinct intracellular insulin- and exercise-recruitable GLUT-4 transporter pools.
急性运动与胰岛素一样,可增加大鼠后肢肌肉对D-葡萄糖的摄取。在此,我们研究了从对照、运动和急性胰岛素处理的大鼠制备的骨骼肌质膜和细胞内膜组分中肌肉葡萄糖转运蛋白GLUT-4和GLUT-1的分布。用抗GLUT-4多克隆抗体进行免疫印迹分析表明,急性胰岛素处理(通过后肢灌注或体内注射)使质膜组分中的GLUT-4转运蛋白增加,而使细胞内膜组分中的GLUT-4转运蛋白减少。运动也增加了质膜中的GLUT-4转运蛋白,但与胰岛素不同的是,运动并未使细胞内组分中的GLUT-4转运蛋白显著减少。用抗GLUT-1抗体进行免疫印迹分析显示,该转运蛋白主要定位于质膜。胰岛素和运动均未显著增加质膜中的GLUT-1转运蛋白。数据表明,GLUT-4是骨骼肌中一种对胰岛素有反应的葡萄糖转运蛋白,此外,GLUT-4也对急性运动有反应。这些结果与GLUT-4葡萄糖转运蛋白从细胞内储存库募集到质膜一致。此外,对运动敏感的GLUT-4转运蛋白并非源自对胰岛素敏感的细胞内膜组分,这表明存在不同的细胞内胰岛素可募集和运动可募集的GLUT-4转运蛋白池。