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Immunocytochemical Detection of GLUT2 at the Rat Intestinal Brush-border Membrane

Julie A. Affleck, Philip A. Helliwell and George L. Kellett

Department of Biology, University of York, York, United Kingdom

Correspondence to: George L. Kellett, Dept. of Biology (Area 3), University of York, York YO10 5YW, UK. E-mail: glk1{at}york.ac.uk

We have proposed a new model of intestinal sugar absorption in which high sugar concentrations promote rapid insertion of the facilitative transporter GLUT2 into the brush-border membrane so that absorptive capacity is precisely regulated to match dietary intake during the assimilation of a meal. However, location of GLUT2 at the brush border by immunocytochemistry has been problematical. We report that control of rapid GLUT2 trafficking and the use of an antibody to a sequence within the large extracellular loop of GLUT2 permits localization of GLUT2 at the brush border. To reveal brush-border GLUT2 fully, it is necessary to digest the sugar chain at the glycosylation site close to the antigenic site. In this way, we have demonstrated by immunocytochemistry PKC-dependent changes in the regulation of brush-border GLUT2 in rat jejunum that correspond to those seen by Western blotting. The functional and immunocytochemical data are now reconciled.

(J Histochem Cytochem 51:1567–1574, 2003)

Key Words: intestine • sugar transport • fructose • GLUT5 • SGLT1

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