Characterization and histochemical localization of the rat intestinal Na(+)-D-glucose cotransporter by monoclonal antibodies.
The localization of the Na(+)-D-glucose cotransporter in rat small intestine was investigated with four monoclonal antibodies which were raised against porcine renal brush-border membrane proteins. The antibodies alter high affinity phlorizin binding or Na+ gradient-dependent D-glucose uptake in kidney and intestine. In both organs, the antibodies react with polypeptides with apparent molecular weights of 75,000 and 47,000. In pig kidney, these polypeptides were identified as components of the Na(+)-D-glucose cotransporter (Koepsell, H., K. Korn, A. Raszeja-Specht, S. Bernotat-Danielowski, D. Ollig, J. Biol. Chem. 263, 18419-18429 (1988)). The electron microscopic localization of antibody binding was investigated by immunogold labeling of ultrathin plastic sections. In villi and crypts of duodenum, jejunum and ileum the antibodies bound specifically to brush-border membranes of enterocytes and did not react with the basolateral membranes. The density of antigenic sites in brush-border membranes was highest in jejunum, intermediate in ileum and lowest in duodenum. On the tip, the middle and the basis of the villi the density of antigenic sites was similar. The data demonstrate homologous Na(+)-D-glucose cotransporters in kidney and intestine. They suggest that during maturation of the enterocytes when the total area of brush-border membrane increases, the concentration of the Na(+)-D-glucose cotransporter in the brush-border membrane remains constant. However, we found that different segments of small intestine not only contain different surface areas of the transporter-containing brush-border membrane per intestinal length but also different densities of the transporter within the brush-border membrane.
Haase
W
W
Max-Planck-Institut für Biophysik, Frankfurt a. M., Bundesrepublik Deutschland.
Heitmann
K
K
Friese
W
W
Ollig
D
D
Koepsell
H
H
D000818:Animals; D000911:Antibodies, Monoclonal; D002462:Cell Membrane / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D006651:Histocytochemistry; D007150:Immunohistochemistry; D007422:Intestines / Q000737:chemistry* / Q000166:cytology; D007668:Kidney / Q000737:chemistry / Q000648:ultrastructure; D008871:Microvilli / Q000737:chemistry; D009004:Monosaccharide Transport Proteins / Q000032:analysis*; D010695:Phlorhizin / Q000378:metabolism; D051381:Rats
D000911:Antibodies, Monoclonal; D009004:Monosaccharide Transport Proteins; D010695:Phlorhizin