Tracking GLUT2 Translocation by Live-Cell Imaging

Methods Mol Biol. 2018:1713:241-254. doi: 10.1007/978-1-4939-7507-5_18.

Abstract

The facilitative glucose transporter (GLUT) family plays a key role in metabolic homeostasis, controlling the absorption rates and rapid response to changing carbohydrate levels. The facilitative GLUT2 transporter is uniquely expressed in metabolic epithelial cells of the intestine, pancreas, liver, and kidney. GLUT2 dysfunction is associated with several pathologies, including Fanconi-Bickel syndrome, a glycogen storage disease, characterized by growth retardation and renal dysfunction. Interestingly, GLUT2 activity is modulated by its cellular localization. Membrane translocation specifically regulates GLUT2 activity in enterocytes, pancreatic β-cells, hepatocytes, and proximal tubule cells. We have established a system to visualize and quantify GLUT2 translocation, and its dynamics, by live imaging of a mCherry-hGLUT2 fusion protein in polarized epithelial cells. This system enables testing of putative modulators of GLUT2 translocation, which are potential drugs for conditions of impaired glucose homeostasis and associated nephropathy.

Keywords: GLUT2; Glucose transport; Kidney function; Live imaging; Polarized epithelium; Translocation; Type 2 diabetes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Dogs
  • Epithelial Cells / metabolism
  • Glucose / metabolism
  • Glucose Transporter Type 2 / metabolism*
  • Humans
  • Kidney / metabolism
  • Madin Darby Canine Kidney Cells
  • Molecular Imaging* / methods
  • Protein Transport

Substances

  • Glucose Transporter Type 2
  • Glucose