TY - JOUR TI - Microtubule-mediated GLUT4 trafficking is disrupted in insulin-resistant skeletal muscle AU - Knudsen, Jonas R AU - Persson, Kaspar W AU - Henriquez-Olguin, Carlos AU - Li, Zhencheng AU - Di Leo, Nicolas AU - Hesselager, Sofie A AU - Raun, Steffen H AU - Hingst, Janne R AU - Trouillon, Raphaël AU - Wohlwend, Martin AU - Wojtaszewski, Jørgen FP AU - Gijs, Martin AM AU - Jensen, Thomas Elbenhardt A2 - Czech, Michael A2 - Akhmanova, Anna A2 - Klip, Amira VL - 12 PY - 2023 DA - 2023/04/19 SP - e83338 C1 - eLife 2023;12:e83338 DO - 10.7554/eLife.83338 UR - https://doi.org/10.7554/eLife.83338 AB - Microtubules serve as tracks for long-range intracellular trafficking of glucose transporter 4 (GLUT4), but the role of this process in skeletal muscle and insulin resistance is unclear. Here, we used fixed and live-cell imaging to study microtubule-based GLUT4 trafficking in human and mouse muscle fibers and L6 rat muscle cells. We found GLUT4 localized on the microtubules in mouse and human muscle fibers. Pharmacological microtubule disruption using Nocodazole (Noco) prevented long-range GLUT4 trafficking and depleted GLUT4-enriched structures at microtubule nucleation sites in a fully reversible manner. Using a perifused muscle-on-a-chip system to enable real-time glucose uptake measurements in isolated mouse skeletal muscle fibers, we observed that Noco maximally disrupted the microtubule network after 5 min without affecting insulin-stimulated glucose uptake. In contrast, a 2-hr Noco treatment markedly decreased insulin responsiveness of glucose uptake. Insulin resistance in mouse muscle fibers induced either in vitro by C2 ceramides or in vivo by diet-induced obesity, impaired microtubule-based GLUT4 trafficking. Transient knockdown of the microtubule motor protein kinesin-1 protein KIF5B in L6 muscle cells reduced insulin-stimulated GLUT4 translocation while pharmacological kinesin-1 inhibition in incubated mouse muscles strongly impaired insulin-stimulated glucose uptake. Thus, in adult skeletal muscle fibers, the microtubule network is essential for intramyocellular GLUT4 movement, likely functioning to maintain an insulin-responsive cell surface recruitable GLUT4 pool via kinesin-1-mediated trafficking. KW - skeletal muscle KW - GLUT4 KW - insulin resistance KW - microtubules KW - kinesin JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -