Knockdown of TRPC3 with siRNA coupled to carbon nanotubes results in decreased insulin-mediated glucose uptake in adult skeletal muscle cells
2009 (English)In: The FASEB Journal, ISSN 0892-6638, E-ISSN 1530-6860, Vol. 23, no 6, 1728-1738 p.Article in journal (Refereed) Published
The involvement of Ca2+ in the insulin-mediated signaling cascade, resulting in glucose uptake in skeletal muscle, is uncertain. Here, we test the hypothesis that Ca2+ influx through canonical transient receptor potential 3 (TRPC3) channels modulates insulin-mediated glucose uptake in adult skeletal muscle. Experiments were performed on adult skeletal muscle cells of wild-type (WT) and obese, insulin-resistant ob/ob mice. Application of the diacylglycerol analog 1-oleyl-2-acetyl-sn-glycerol (OAG) induced a nonselective cation current, which was inhibited by the addition of anti-TRPC3 antibody in the patch pipette and smaller in ob/ob than in WT cells. Knockdown of TRPC3, using a novel technique based on small interfering RNA (siRNA) coupled to functionalized carbon nanotubes, resulted in pronounced (similar to 70%) decreases in OAG-induced Ca2+ influx and insulin-mediated glucose uptake. TRPC3 and the insulin-sensitive glucose transporter 4 (GLUT4) coimmunoprecipitated, and immunofluorescence staining showed that they were colocalized in the proximity of the transverse tubular system, which is the predominant site of insulin-mediated glucose transport in skeletal muscle. In conclusion, our results indicate that TRPC3 interacts functionally and physically with GLUT4, and Ca2+ influx through TRPC3 modulates insulin-mediated glucose uptake. Thus, TRPC3 is a potential target for treatment of insulin-resistant conditions.-Lanner, J. T., Bruton, J. D., Assefaw-Redda, Y., Andronache, Z., Zhang, S.- J., Severa, D., Zhang, Z.- B., Melzer, W., Zhang, S.-L., Katz, A., Westerblad, H. Knockdown of TRPC3 with siRNA coupled to carbon nanotubes results in decreased insulin-mediated glucose uptake in adult skeletal muscle cells. FASEB J. 23, 1728-1738 (2009)
Place, publisher, year, edition, pages
2009. Vol. 23, no 6, 1728-1738 p.
Ca2+, sarcolemma, transverse tubules, near-membrane Ca2+, colocalization, protein-kinase-c, stimulated glut4 translocation, cation channels, transverse tubules, plasma-membrane, calcium sensor, vesicle fusion, slow-twitch, ca2+ influx, activation
IdentifiersURN: urn:nbn:se:kth:diva-18486DOI: 10.1096/fj.08-116814ISI: 000266652400013ScopusID: 2-s2.0-67649359890OAI: oai:DiVA.org:kth-18486DiVA: diva2:336533
QC 201005252010-08-052010-08-052011-01-31Bibliographically approved