Ceramide- and Oxidant-Induced Insulin Resistance Involve Loss of Insulin-Dependent Rac-Activation and Actin Remodeling in Muscle Cells

  1. Lellean JeBailey12,
  2. Oshrit Wanono1,
  3. Wenyan Niu1,
  4. Jessica Roessler1,
  5. Assaf Rudich1 and
  6. Amira Klip12
  1. 1Programme in Cell Biology, the Hospital for Sick Children, Toronto, Ontario, Canada
  2. 2Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada
  1. Address correspondence and reprint requests to Dr. Amira Klip, Programme in Cell Biology, The Hospital for Sick Children, 555 University Ave., Toronto, ON, Canada M5G 1X8. E-mail: amira{at}sickkids.ca


In muscle cells, insulin elicits recruitment of the glucose transporter GLUT4 to the plasma membrane. This process engages sequential signaling from insulin receptor substrate (IRS)-1 to phosphatidylinositol (PI) 3-kinase and the serine/threonine kinase Akt. GLUT4 translocation also requires an Akt-independent but PI 3-kinase–and Rac-dependent remodeling of filamentous actin. Although IRS-1 phosphorylation is often reduced in insulin-resistant states in vivo, several conditions eliciting insulin resistance in cell culture spare this early step. Here, we show that insulin-dependent Rac activation and its consequent actin remodeling were abolished upon exposure of L6 myotubes beginning at doses of C2-ceramide or oxidant-producing glucose oxidase as low as 12.5 μmol/l and 12.5 mU/ml, respectively. At 25 μmol/l and 25 mU/ml, glucose oxidase and C2-ceramide markedly reduced GLUT4 translocation and glucose uptake and lowered Akt phosphorylation on Ser473 and Thr308, yet they affected neither IRS-1 tyrosine phosphorylation nor its association with p85 and PI 3-kinase activity. Small interfering RNA–dependent Rac1 knockdown prevented actin remodeling and GLUT4 translocation but spared Akt phosphorylation, suggesting that Rac and actin remodeling do not contribute to overall Akt activation. We propose that ceramide and oxidative stress can each affect two independent arms of insulin signaling to GLUT4 at distinct steps, Rac–GTP loading and Akt phosphorylation.


  • W.N. is currently affiliated with the Department of Immunology, Tianjin Medical University, Tianjin, China. A.R. is currently affiliated with the Department of Clinical Biochemistry and the S. Daniel Abraham Center for Health and Nutrition, Ben Gurion University, Beer Sheva, Israel.

    Additional information can be found in an online appendix at http://dx.doi.org/10.2337/db06-0823.

    The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

    • Accepted October 19, 2006.
    • Received June 16, 2006.
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