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Fiber type effects on contraction-stimulated glucose uptake and GLUT4 abundance in single fibers from rat skeletal muscle

机译:纤维类型对大鼠骨骼肌单纤维收缩刺激葡萄糖摄取和GLUT4丰度的影响

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摘要

To fully understand skeletal muscle at the cellular level, it is essential to evaluate single muscle fibers. Accordingly, the major goals of this study were to determine if there are fiber type-related differences in single fibers from rat skeletal muscle for: 1) contraction-stimulated glucose uptake and/or 2) the abundance of GLUT4 and other metabolically relevant proteins. Paired epitrochlearis muscles isolated from Wistar rats were either electrically stimulated to contract (E-Stim) or remained resting (No E-Stim). Single fibers isolated from muscles incubated with 2-deoxy-d-[3H]glucose (2-DG) were used to determine fiber type [myosin heavy chain (MHC) isoform protein expression], 2-DG uptake, and abundance of metabolically relevant proteins, including the GLUT4 glucose transporter. E-Stim, relative to No E-Stim, fibers had greater (P < 0.05) 2-DG uptake for each of the isolated fiber types (MHC-IIa, MHC-IIax, MHC-IIx, MHC-IIxb, and MHC-IIb). However, 2-DG uptake for E-Stim fibers was not significantly different among these five fiber types. GLUT4, tethering protein containing a UBX domain for GLUT4 (TUG), cytochrome c oxidase IV (COX IV), and filamin C protein levels were significantly greater (P < 0.05) in MHC-IIa vs. MHC-IIx, MHC-IIxb, or MHC-IIb fibers. TUG and COX IV in either MHC-IIax or MHC-IIx fibers exceeded values for MHC-IIxb or MHC-IIb fibers. GLUT4 levels for MHC-IIax fibers exceeded MHC-IIxb fibers. GLUT4, COX IV, filamin C, and TUG abundance in single fibers was significantly (P < 0.05) correlated with each other. Differences in GLUT4 abundance among the fiber types were not accompanied by significant differences in contraction-stimulated glucose uptake.
机译:为了在细胞水平上充分了解骨骼肌,评估单个肌肉纤维是必不可少的。因此,本研究的主要目标是确定大鼠骨骼肌单纤维中是否存在与纤维类型相关的差异:1)收缩刺激的葡萄糖摄取和/或2)GLUT4和其他代谢相关蛋白的丰度。从Wistar大鼠中分离出的成对上棘肌被电刺激收缩(E-Stim)或保持静止(无E-Stim)。用2-脱氧-d-[ 3 H]葡萄糖(2-DG)孵育的肌肉分离出的单纤维用于确定纤维类型[肌球蛋白重链(MHC)同工型蛋白表达],2- DG的摄取,以及包括GLUT4葡萄糖转运蛋白在内的代谢相关蛋白质的丰度。相对于无E-Stim,E-Stim纤维对每种分离的纤维类型(MHC-IIa,MHC-IIax,MHC-IIx,MHC-IIxb和MHC- IIb)。但是,在这五种纤维类型中,E-Stim纤维的2-DG吸收量没有显着差异。与MHC-IIx,MHC-IIxb,MHC-IIx,MHC-IIx,MHC-IIx,MHC-IIa中含有GLUT4(TUG)UBX域的束缚蛋白(TUG),细胞色素C氧化酶IV(COX IV)和纤维蛋白C蛋白水平相比,GLUT4显着更高(P <0.05)。或MHC-IIb纤维。 MHC-IIax或MHC-IIx纤维中的TUG和COX IV超过MHC-IIxb或MHC-IIb纤维的值。 MHC-IIax纤维的GLUT4水平超过MHC-IIxb纤维。单纤维中的GLUT4,COX IV,纤维蛋白C和TUG的丰度显着相关(P <0.05)。纤维类型之间GLUT4丰度的差异并没有伴随收缩刺激的葡萄糖摄取的显着差异。

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