Immunohistochemical characterization of a differentiation-specific carbohydrate epitope in the plasma membrane of the epithelial cells of rat small intestine.

Abstract

The monoclonal antibody (MAb) SI/EC1 was produced by immunization of Balb/c mice with an antigen prepared from the isolated basolateral membrane (BLM) of rat small intestine epithelial cells by trypsin cleavage. Immunohistochemical labeling at the light and electron microscopic level shows that the SI/EC1 epitope is localized in the plasma membrane (PM) of the small intestine epithelial cells and is expressed around Day 21 after birth (weaning time). There are, however, differences in the labeling between crypt and villous cells. In the crypt cells, the microvillous membrane (MVM) and the lateral part of the BLM are strongly labeled, whereas the basal part of the BLM is unlabeled. In the villous cells, both the MVM and the basal and lateral part of the BLM are labeled, but the labeling is not as intense as in the crypts. In immunoblotting experiments with the isolated BLM, three protein bands (125 KD, 110 KD, and 90 KD) were labeled specifically with the MAb. Enzymic cleaving of the BLM with exo- and endoglycosidases and subsequent immunoblotting, as well as other findings, suggest that the specific structure of the SI/EC1 epitope consists mainly of carbohydrates (CH) (oligosaccharides). This finding points out the possibility that this epitope may have something to do with the variable adhesion of the small intestine epithelial cells along the crypt-villus axis.