Douen A G, Ramlal T, Klip A, Young D A, Cartee G D, Holloszy J O
Division of Cell Biology, Hospital for Sick Children, Toronto, Ontario, Canada.
Endocrinology. 1989 Jan;124(1):449-54. doi: 10.1210/endo-124-1-449.
A previously developed technique for the isolation of plasma and intracellular membrane fractions from rat skeletal muscle was used to investigate transporter migration after insulin treatment or a bout of exercise (45 min of treadmill). Glucose-inhibitable cytochalasin-B binding was used to estimate the number of glucose transporters. Insulin and exercise caused increases in glucose uptake into the hindlimb muscles of 5- and 3-fold, respectively. Each stimulus also caused a 2-fold increase in the number of glucose transporters in plasma membranes prepared from hindlimb muscles. The insulin-induced increase in plasma membrane transporters was accompanied by a concomitant decrease in transporters from the intracellular pool. In contrast to insulin, there was no concomitant decrease in the number of cytochalasin-B-binding sites in the intracellular membrane fraction from exercised muscles. The ability of both insulin and exercise to increase the number of transporters in the plasma membrane is in accordance with recruitment of transporters as one cause of increased transport activity. However, the inability of exercise to decrease the number of transporters in the insulin-sensitive intracellular pool suggests the existence of either a second recruitable transporter pool or masked glucose transporters in the plasma membrane that are unmasked by the muscle contractile activity.
采用先前开发的从大鼠骨骼肌中分离血浆和细胞内膜组分的技术,来研究胰岛素处理或一次运动(跑步机运动45分钟)后转运体的迁移情况。利用葡萄糖抑制的细胞松弛素B结合来估计葡萄糖转运体的数量。胰岛素和运动分别使后肢肌肉的葡萄糖摄取增加了5倍和3倍。每种刺激还使从后肢肌肉制备的质膜中葡萄糖转运体的数量增加了2倍。胰岛素诱导的质膜转运体增加伴随着细胞内池转运体的相应减少。与胰岛素不同,运动肌肉的细胞内膜组分中细胞松弛素B结合位点的数量没有相应减少。胰岛素和运动增加质膜中转运体数量的能力与转运体的募集一致,这是转运活性增加的一个原因。然而,运动不能减少胰岛素敏感的细胞内池中转运体的数量,这表明存在第二个可募集的转运体池,或者质膜中存在被肌肉收缩活动解开掩盖的葡萄糖转运体。