Unit

UNIT 15.15 Documenting GLUT4 Exocytosis and Endocytosis in Muscle Cell Monolayers

  1. Shuhei Ishikura,
  2. Costin N. Antonescu,
  3. Amira Klip

Published Online: 1 MAR 2010

DOI: 10.1002/0471143030.cb1515s46

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Ishikura, S., Antonescu, C. N. and Klip, A. 2010. Documenting GLUT4 Exocytosis and Endocytosis in Muscle Cell Monolayers. Current Protocols in Cell Biology. 46:15.15:15.15.1–15.15.19.

Author Information

  1. Program in Cell Biology, The Hospital for Sick Children, Toronto, Ontario, Canada

Publication History

  1. Published Online: 1 MAR 2010
  2. Published Print: MAR 2010

Abstract

The elevated blood glucose following a meal is cleared by insulin-stimulated glucose entry into muscle and fat cells. The hormone increases the amount of the glucose transporter GLUT4 at the plasma membrane in these tissues at the expense of preformed intracellular pools. In addition, muscle contraction also increases glucose uptake via a gain in GLUT4 at the plasma membrane. Regulation of GLUT4 levels at the cell surface could arise from alterations in the rate of its exocytosis, endocytosis, or both. Hence, methods that can independently measure these traffic parameters for GLUT4 are essential to understanding the mechanism of regulation of membrane traffic of the transporter. Here, we describe cell population–based assays to measure the steady-state levels of GLUT4 at the cell surface, as well as to separately measure the rates of GLUT4 endocytosis and endocytosis. Curr. Protoc. Cell Biol. 46:15.15.1-15.15.19. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • glucose;
  • internalization;
  • insulin;
  • muscle contraction;
  • regulated membrane traffic