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Experimental Mechanics

, Volume 49, Issue 1, pp 3–9 | Cite as

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

  • S. R. K. Vedula
  • T. S. Lim
  • P. J. Kausalya
  • E. B. Lane
  • G. Rajagopal
  • W. Hunziker
  • C. T. Lim
Article

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.

Keywords

Intercellular adhesion Tight junctions Dual micropipette assay Claudin Occludin Actin 

Notes

Acknowledgement

The authors would like to acknowledge funding support provided by the Biomedical Research Council (BMRC) from the Agency for Science, Technology and Research (A*STAR).

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Copyright information

© Society for Experimental Mechanics 2007

Authors and Affiliations

  • S. R. K. Vedula
    • 1
  • T. S. Lim
    • 2
  • P. J. Kausalya
    • 3
  • E. B. Lane
    • 4
  • G. Rajagopal
    • 5
  • W. Hunziker
    • 3
  • C. T. Lim
    • 1
    • 6
  1. 1.Nano Biomechanics Lab, Division of BioengineeringNational University of SingaporeSingaporeSingapore
  2. 2.Bioinformatics InstituteSingaporeSingapore
  3. 3.Institute of Molecular and Cell BiologySingaporeSingapore
  4. 4.Institute of Medical BiologySingaporeSingapore
  5. 5.The Cancer Institute of New JerseyNew BrunswickUSA
  6. 6.Division of Bioengineering & Department of Mechanical EngineeringNational University of SingaporeSingaporeSingapore

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