Quantifying Forces Mediated by Integral Tight Junction Proteins in Cell–Cell Adhesion

Abstract

Cellular adhesion and barriers formed by intercellular adhesion proteins [tight junctions (occludin and claudins) and adherens junction (E-cadherin)] are important in maintaining tissue homeostasis. However, disruption of these junction proteins is associated with diseases in the organ systems such as multiple sclerosis, diarrhea, asthma, and gastro-intestinal tract carcinomas among others. In this paper, the separation force needed to separate two cells expressing some of these proteins was measured using the dual micropipette assay. Results show that L-fibroblasts transfected with claudin-1 and claudin-2 exhibit higher separation force (~2.8 nN and 2.3 nN, respectively) as compared to control cells or cells transfected with occludin (~1 nN). Furthermore, the separation force was not affected on addition of calcium chelating agent (ethylene diamine tetra acetic acid, EDTA). The separation force was, however, significantly decreased on treating cells with the actin disrupting agent Cytochalasin-D. These results show that the dual micropipette assay is a simple and useful experimental technique for quantifying cell–cell adhesion.