Role of PKCΖ translocation in the development of type 2 diabetes in rats following continuous glucose infusion

Abstract

Aim We investigated the molecular mechanisms of hyperglycaemia-induced insulin resistance and type 2 diabetes in rats receiving a continuous glucose infusion (GI). Methods Female Wistar rats were infused with either 2.8 mol/L glucose or saline (2 mL/h) for durations varying from 0 to 15 days. Blood samples were analysed daily to determine glucose and insulin dynamics. Subsets of animals were sacrificed and solues muscles were extracted for determination of protein expression, subcellular location, and activities of insulin-signalling proteins. Results Rats accommodated this systemic glucose oversupply and developed insulin resistance on day 5 (normoglycaemia/hyperinsulinaemia) and type 2 diabetes on day 15 (hyperglycaemia/normoinsulinaemia). The effect of GI on protein kinase CΖ (PKCΖ) activity was independent of changes in phosphatidylinositol 3-kinase activity, and occurred in parallel with an increase in PDK1 activity. Activated PKCΖ was mainly located in the cytosol after 5 days of GI that was coincident with the translocation of GLUT4 to the plasma membrane, and normoglycaemia. After 15 days of GI, PKCΖ translocated from the cytosol to the plasma membrane with a concomitant decrease in PDK1 activity. This caused an increase in the association between PKCΖ and PKB and a decrease in PDK1–PKB reactions at the plasma membrane, leading to reduced PKB activity. The activity of PKCΖ per se was also compromised. The PKCΖ and PKB activity reduction and the blunted insulin-stimulated GLUT4 translocation eventually led to hyperglycaemia and diabetes. Conclusion Translocation of PKCΖ may play a central role in the development of type 2 diabetes. Copyright © 2009 John Wiley & Sons, Ltd.