Claudins pp 27-41 | Cite as

Electrophysiological Characterization of Claudin Ion Permeability Using Stably Transfected Epithelial Cell Lines

  • Alan S. L. YuEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 762)


Claudins are tight junction membrane proteins that act as paracellular pores and barriers and regulate epithelial permeability to small ions. A key step in understanding the function of any claudin isoform is the in vitro measurement of its ion permeability and selectivity. Herein, we describe methods to generate clonal lines with stable inducible overexpression of claudins in Madin–Darby canine kidney epithelial cells, measure conductance and diffusion potentials in Ussing chambers, correct for liquid junction potentials, and derive quantitatively accurate values for individual ion permeabilities.

Key words

Claudin Tight junction Paracellular Diffusion potential Conductance Ion permeability Junction potential 


  1. 1.
    Tsukita S, Furuse M (2002) Claudin-based barrier in simple and stratified cellular sheets. Curr Opin Cell Biol 14:531–536PubMedCrossRefGoogle Scholar
  2. 2.
    Van Itallie CM, Anderson JM (2006) Claudins and epithelial paracellular transport. Annu Rev Physiol 68:403–429PubMedCrossRefGoogle Scholar
  3. 3.
    Angelow S, Ahlstrom R, Yu AS (2008) Biology of claudins. Am J Physiol Renal Physiol 295:F867–876PubMedCrossRefGoogle Scholar
  4. 4.
    Saier MH, Jr. (1981) Growth and differentiated properties of a kidney epithelial cell line (MDCK). Am J Physiol 240:C106–109PubMedGoogle Scholar
  5. 5.
    Cereijido M, Gonzalez-Mariscal L, Borboa L (1983) Occluding junctions and paracellular pathways studied in monolayers of MDCK cells. J Exp Biol 106:205–215PubMedGoogle Scholar
  6. 6.
    Van Itallie C, Fanning AS, Anderson JM (2003) Reversal of charge selectivity in cation or anion selective epithelial lines by expression of different claudins. Am J Physiol Cell Physiol 286:F1078–1084CrossRefGoogle Scholar
  7. 7.
    Angelow S, Schneeberger EE, Yu AS (2007) Claudin-8 expression in renal epithelial cells augments the paracellular barrier by replacing endogenous claudin-2. J Membr Biol 215:147–159PubMedCrossRefGoogle Scholar
  8. 8.
    Gossen M, Bujard H (1992) Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc Natl Acad Sci U S A 89:5547–5551PubMedCrossRefGoogle Scholar
  9. 9.
    Barry PH, Diamond JM (1970) Junction potentials, electrode standard potentials, and other problems in interpreting electrical properties of membranes. J Membr Biol 3:93–122CrossRefGoogle Scholar
  10. 10.
    Yu AS, Cheng MH, Angelow S, et al. (2009) Molecular basis for cation selectivity in claudin-2-based paracellular pores: identification of an electrostatic interaction site. J Gen Physiol 133:111–127PubMedCrossRefGoogle Scholar
  11. 11.
    Hille B (2001) Ionic channels of excitable membranes. Sunderland: Sinauer.Google Scholar
  12. 12.
    Kimizuka H, Koketsu K (1964) Ion transport through cell membrane. J Theoret Biol 6:290–305CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Division of Nephrology and Hypertension and the Kidney InstituteUniversity of Kansas Medical CenterKansas CityUSA

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