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Knockdown of TRPC3 with siRNA coupled to carbon nanotubes results in decreased insulin‐mediated glucose uptake in adult skeletal muscle cells
Author(s) -
Lanner Johanna T.,
Bruton Joseph D.,
Assefaw-Redda Yohannes,
Andronache Zoita,
Zhang Shi-Jin,
Severa Denise,
Zhang Zhi-Bin,
Melzer Werner,
Zhang Shi-Li,
Katz Abram,
Westerblad Håkan
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.08-116814
Subject(s) - trpc3 , glut4 , insulin , glucose uptake , glucose transporter , skeletal muscle , gene knockdown , medicine , insulin receptor , chemistry , endocrinology , microbiology and biotechnology , transient receptor potential channel , biology , biochemistry , insulin resistance , receptor , trpc , apoptosis
The involvement of Ca 2+ in the insulin‐mediated signaling cascade, resulting in glucose uptake in skeletal muscle, is uncertain. Here, we test the hypothesis that Ca 2+ 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 (~70%) decreases in OAG‐induced Ca 2+ 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 Ca 2+ 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)