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JHC exPRESS: First Published March 19, 2007. doi:10.1369/jhc.6A7165.2007
Journal of Histochemistry and Cytochemistry
Copyright © 2007 Tokunaga et al.

A more recent version of this article appeared on July 1, 2007.
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Articles

A Novel Monoclonal Antibody Against Second Extracellular Loop of Occludin Disrupts Epithelial Cell Polarity

Yuichi Tokunaga 1, Takashi Kojima 1*, Makoto Osanai 1, Masaki Murata 1, Hideki Chiba 1, Hirotoshi Tobioka 1 and Norimasa Sawada 1

1 Department of Pathology, Sapporo Medical University School of Medicine, Sapporo, Japan (YT,TK,MO,MM,HC,HT,NS); Sapporo Clinical Laboratory Co., Sapporo, Japan (YT); and Otaru Kyoukai Hospital, Otaru, Japan (HT)

* To whom correspondence should be addressed. E-mail: ktakashi{at}sapmed.ac.jp.

Submitted on December 5, 2006
Accepted on 1 March 2007


  Abstract
The tight junction (TJ) regulates epithelial cell polarity and paracellular permeability. In the present study, to investigate whether the second extracellular loop of occludin affects the localization of CEA and CD26 expressed on apical membranes, and the fence function of the TJ, the human intestinal epithelial cell line T84 was treated with the monoclonal anti-occludin antibody (mAb) 1H8, corresponding to the second extracellular loop of occludin. In T84 cells treated with mAb 1H8, occludin disappeared, and CEA and CD26 were observed to diffuse from the apical membrane to the basolateral membrane. Furthermore, a decrease in the fence function of TJ was observed without changes in the TJ strands and barrier function. When T84 cells precultured in low Ca medium were re-cultured in normal Ca medium in the presence of mAb 1H8, the recruitment of occludin to apicalmost membranes and recovery in distribution of CEA and CD26 were markedly retarded compared to the control. These results suggested that mAb 1H8 against the second extracellular loop of occludin selectively affected formation of the apical/basolateral intramembrane diffusion barrier and that the second extracellular loop of occludin plays a crucial role in the maintenance of epithelial cell polarity by the TJ.

Key Words: occludin, epithelial cell polarity, monoclonal antibody, CEA, CD26


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