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Adipocyte and Cell Biology

Temporal analysis of mechanisms leading to stimulation of glucose uptake in skeletal muscle cells by an adipokine mixture derived from primary rat adipocytes

International Journal of Obesity volume 35, pages 355–363 (2011)Cite this article

Subjects

Abstract

Objective:

The direct effects of adipokines on skeletal muscle metabolism have been well established. As the combinatorial effects of adipokine mixtures are likely to be of more physiological relevance, we used a coculture system of primary rat adipocytes and L6 skeletal muscle cells to examine the effects of adiponectin derived from primary rat adipocytes on rat skeletal muscle cells.

Results:

We showed that coculture with adipocytes stimulated glucose uptake in L6 cells within 30 min and this correlated with an increase of glucose transporter isoform 4 (GLUT4) localization to the plasma membrane. These effects were dependent on the reorganization of the actin cytoskeleton, demonstrated by rhodamine-labeled phalloidin immunofluorescence, as cytochalasin D attenuated the glucose uptake induced by adipocyte-conditioned media. Temporal analysis revealed that enhanced glucose uptake was maintained after 24 h of coculture, and this was attributed to an increase in both GLUT1 expression and the cell surface content of GLUT4. We established a role for adiponectin in mediating these effects as antibody-mediated neutralization attenuated the metabolic effects of adipocyte-conditioned media. Furthermore, compound C blocked these effects, suggesting an important role for AMPK. Importantly, when we compared the effects of full-length recombinant adiponectin with adipocyte-conditioned media, we confirmed that recombinant adiponectin was unable to stimulate glucose uptake in L6 cells despite having an important role in adipocyte-conditioned media.

Conclusions:

Our results demonstrate the importance of examining the effects of adipokines in the context of physiologically relevant mixtures to accurately determine their metabolic effects on skeletal muscle.

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Acknowledgements

Funding was provided by the Canadian Institutes of Health Research via an operating Grant and the New Investigator award to GS, VV supported by a Doctoral Student Research Award from the Canadian Diabetes Association. XZ received support as a China Scholarship Council Visiting Scholar.

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Authors and Affiliations

  1. Department of Biology, York University, Toronto, Canada

    V Vu, K Dadson, T Odisho, W Kim, X Zhou, F Thong & G Sweeney

  2. Institut Pasteur Korea, Seoul, South Korea

    G Sweeney

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  1. V Vu
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  2. K Dadson
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Correspondence to G Sweeney.

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Vu, V., Dadson, K., Odisho, T. et al. Temporal analysis of mechanisms leading to stimulation of glucose uptake in skeletal muscle cells by an adipokine mixture derived from primary rat adipocytes. Int J Obes 35, 355–363 (2011). https://doi.org/10.1038/ijo.2010.160

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