Microtubule-mediated GLUT4 trafficking is disrupted in insulin resistant skeletal muscle

  1. Jonas R Knudsen  Is a corresponding author
  2. Kaspar W Persson
  3. Carlos Henriquez-Olguin
  4. Zhencheng Li
  5. Nicolas Di Leo
  6. Sofie A Hesselager
  7. Steffen H Raun
  8. Janne R Hingst
  9. Raphaël Trouillon
  10. Martin Wohlwend
  11. Jørgen FP Wojtaszewski
  12. Martin AM Gijs
  13. Thomas Elbenhardt Jensen  Is a corresponding author
  1. University of Copenhagen, Denmark
  2. Novo Nordisk, Denmark
  3. Polytechnique Montréal, Canada
  4. École Polytechnique Fédérale de Lausanne, Switzerland

Abstract

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 2h 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.

Data availability

All data generated or analyzed during this study are included in the manuscript and supporting files

Article and author information

Author details

  1. Jonas R Knudsen

    Department of Nutrition, Exercise and Sports, University of Copenhagen, Copenhagen, Denmark
    For correspondence
    jrk@nexs.ku.dk
    Competing interests
    Jonas R Knudsen, Affiliated with Novo Nordisk A/S.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5471-491X
  2. Kaspar W Persson

    Department of Nutrition, Exercise and Sports, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    No competing interests declared.
  3. Carlos Henriquez-Olguin

    Department of Nutrition, Exercise and Sports, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    No competing interests declared.
  4. Zhencheng Li

    Department of Nutrition, Exercise and Sports, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    No competing interests declared.
  5. Nicolas Di Leo

    Department of Nutrition, Exercise and Sports, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6268-890X
  6. Sofie A Hesselager

    Department of Nutrition, Exercise and Sports, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    No competing interests declared.
  7. Steffen H Raun

    Department of Biomedical Sciences, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    No competing interests declared.
  8. Janne R Hingst

    Clinical Drug Development, Novo Nordisk, Soeborg, Denmark
    Competing interests
    Janne R Hingst, Affiliated with Novo Nordisk A/S.
  9. Raphaël Trouillon

    Department of Electrical Engineering, Polytechnique Montréal, Montreal, Canada
    Competing interests
    No competing interests declared.
  10. Martin Wohlwend

    Institute of Bioengineering, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland
    Competing interests
    No competing interests declared.
  11. Jørgen FP Wojtaszewski

    Department of Nutrition, Exercise and Sports, University of Copenhagen, Copenhagen, Denmark
    Competing interests
    Jørgen FP Wojtaszewski, has ongoing collaborations with Pfizer inc. and Novo Nordisk A/S unrelated to this study..
  12. Martin AM Gijs

    Laboratory of Microsystems, École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland
    Competing interests
    No competing interests declared.
  13. Thomas Elbenhardt Jensen

    Department of Nutrition, Exercise and Sports, University of Copenhagen, Copenhagen, Denmark
    For correspondence
    tejensen@nexs.ku.dk
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6139-8268

Funding

Novo Nordisk Fonden (15182)

  • Thomas Elbenhardt Jensen

Novo Nordisk Fonden (16OC0023046)

  • Jørgen FP Wojtaszewski

Novo Nordisk Fonden (17SA0031406)

  • Jonas R Knudsen

Novo Nordisk Fonden (17SA0031406)

  • Carlos Henriquez-Olguin

Lundbeckfonden (R313-2019-643)

  • Thomas Elbenhardt Jensen

Lundbeckfonden (R266-2017-4358)

  • Jørgen FP Wojtaszewski

Sundhed og Sygdom, Det Frie Forskningsråd (FSS8020-00288B)

  • Jørgen FP Wojtaszewski

Sundhed og Sygdom, Det Frie Forskningsråd (#9058-00047B)

  • Jonas R Knudsen

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Michael Czech, University of Massachusetts Medical School, United States

Ethics

Animal experimentation: All animal experiments were approved by the Danish Animal Experimental Inspectorate or by the local animal experimentation committee of the Canton de Vaud under license 2890 and complied with the European Union legislation as outlined by the European Directive 2010/63/EU. The current work adheres to the standards outlined in the ARRIVE reporting guidelines.

Human subjects: The work involving human subjects was approved by the Copenhagen Ethics Committee (H-6-2014-038; Copenhagen, Denmark) and complied with the guidelines of the 2013 Declaration of Helsinki. Informed written consent was obtained from all subjects prior to entering the study.

Version history

  1. Received: September 8, 2022
  2. Preprint posted: September 22, 2022 (view preprint)
  3. Accepted: April 2, 2023
  4. Accepted Manuscript published: April 19, 2023 (version 1)
  5. Version of Record published: May 10, 2023 (version 2)

Copyright

© 2023, Knudsen et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Jonas R Knudsen
  2. Kaspar W Persson
  3. Carlos Henriquez-Olguin
  4. Zhencheng Li
  5. Nicolas Di Leo
  6. Sofie A Hesselager
  7. Steffen H Raun
  8. Janne R Hingst
  9. Raphaël Trouillon
  10. Martin Wohlwend
  11. Jørgen FP Wojtaszewski
  12. Martin AM Gijs
  13. Thomas Elbenhardt Jensen
(2023)
Microtubule-mediated GLUT4 trafficking is disrupted in insulin resistant skeletal muscle
eLife 12:e83338.
https://doi.org/10.7554/eLife.83338

Share this article

https://doi.org/10.7554/eLife.83338

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