TY - JOUR
T1 - Protein kinase Cδ mediates insulin-induced glucose transport in primary cultures of rat skeletal muscle
AU - Braiman, Liora
AU - Alt, Addy
AU - Kuroki, Toshio
AU - Ohba, Motoi
AU - Bak, Asia
AU - Tennenbaum, Tamar
AU - Sampson, Sanford R.
PY - 1999/12
Y1 - 1999/12
N2 - Insulin activates certain protein kinase C (PKC) isoforms that are involved in insulin-induced glucose transport. In this study, we investigated the possibility that activation of PKCδ by insulin participates in the mediation of insulin effects on glucose transport in skeletal muscle. Studies were performed on primary cultures of rat skeletal myotubes. The role of PKCδ in insulin-induced glucose uptake was evaluated both by selective pharmacological blockade and by over-expression of wild-type and point-mutated inactive PKCδ isoforms in skeletal myotubes. We found that insulin induces tyrosine phosphorylation and translocation of PKCδ to the plasma membrane and increases the activity of this iso-form. Insulin-induced effects on translocation and phosphorylation of PKCδ were blocked by a low concentration of rottlerin, whereas the effects of insulin on other PKC isoforms were not. This selective blockade of PKCδ by rottlerin also inhibited insulin-induced translocation of glucose transporter 4 (GLUT4), but not glucose transporter 3 (GLUT3), and significantly reduced the stimulation of glucose uptake by insulin. When overexpressed in skeletal muscle, PKCδ and PKCα were both active. Overexpression of PKCδ induced the translocation of GLUT4 to the plasma membrane and increased basal glucose uptake to levels attained by insulin. Moreover, insulin did not increase glucose uptake further in cells overexpressing PKCδ. Overexpression of PKCα did not affect basal glucose uptake or GLUT4 location. Stimulation of glucose uptake by insulin in cells overexpressing PKCα was similar to that in untransfected cells. Transfection of skeletal myotubes with dominant negative mutant PKCδ did not alter basal glucose uptake but blocked insulin-induced GLUT4 translocation and glucose transport. These results demonstrate that insulin activates PKCδ and that activated PKCδ is a major signaling molecule in insulin-induced glucose transport.
AB - Insulin activates certain protein kinase C (PKC) isoforms that are involved in insulin-induced glucose transport. In this study, we investigated the possibility that activation of PKCδ by insulin participates in the mediation of insulin effects on glucose transport in skeletal muscle. Studies were performed on primary cultures of rat skeletal myotubes. The role of PKCδ in insulin-induced glucose uptake was evaluated both by selective pharmacological blockade and by over-expression of wild-type and point-mutated inactive PKCδ isoforms in skeletal myotubes. We found that insulin induces tyrosine phosphorylation and translocation of PKCδ to the plasma membrane and increases the activity of this iso-form. Insulin-induced effects on translocation and phosphorylation of PKCδ were blocked by a low concentration of rottlerin, whereas the effects of insulin on other PKC isoforms were not. This selective blockade of PKCδ by rottlerin also inhibited insulin-induced translocation of glucose transporter 4 (GLUT4), but not glucose transporter 3 (GLUT3), and significantly reduced the stimulation of glucose uptake by insulin. When overexpressed in skeletal muscle, PKCδ and PKCα were both active. Overexpression of PKCδ induced the translocation of GLUT4 to the plasma membrane and increased basal glucose uptake to levels attained by insulin. Moreover, insulin did not increase glucose uptake further in cells overexpressing PKCδ. Overexpression of PKCα did not affect basal glucose uptake or GLUT4 location. Stimulation of glucose uptake by insulin in cells overexpressing PKCα was similar to that in untransfected cells. Transfection of skeletal myotubes with dominant negative mutant PKCδ did not alter basal glucose uptake but blocked insulin-induced GLUT4 translocation and glucose transport. These results demonstrate that insulin activates PKCδ and that activated PKCδ is a major signaling molecule in insulin-induced glucose transport.
UR - http://www.scopus.com/inward/record.url?scp=0033233463&partnerID=8YFLogxK
U2 - 10.1210/mend.13.12.0393
DO - 10.1210/mend.13.12.0393
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C2 - 10598577
AN - SCOPUS:0033233463
SN - 0888-8809
VL - 13
SP - 2002
EP - 2012
JO - Molecular Endocrinology
JF - Molecular Endocrinology
IS - 12
ER -