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J Appl Physiol 98: 138-143, 2005. First published July 23, 2004; doi:10.1152/japplphysiol.00478.2004
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Estimation of paracellular conductance of primary rat alveolar epithelial cell monolayers

Kwang-Jin Kim,1,4,7 Zea Borok,1,5,7 Carsten Ehrhardt,9 Brigham C. Willis,8 Claus-Michael Lehr,9 and Edward D. Crandall1,6,7

Departments of 1Medicine, 2Physiology and Biophysics, 3Molecular Pharmacology and Toxicology, 4Biomedical Engineering, 5Biochemistry and Molecular Biology, and 6Pathology, 7Will Rogers Institute Pulmonary Research Center, and 8Department of Anesthesiology Critical Care Medicine, Childrens Hospital of Los Angeles, University of Southern California, Los Angeles, California; and 9Department of Biopharmaceutics and Pharmaceutical Technology, Saarland University, Saarbruecken, Germany

Submitted 5 May 2004 ; accepted in final form 21 July 2004

Freshly isolated rat type II pneumocytes, when grown on permeable tissue culture-treated polycarbonate filters, form confluent alveolar epithelial cell monolayers (RAECM). Cells in RAECM undergo transdifferentiation, exhibiting over time morphological and phenotypic characteristics of type I pneumocytes in vivo. We recently reported that transforming growth factor-{beta}1 (TGF-{beta}1) decreases overall monolayer resistance (Rte) and stimulates short-circuit current in a dose-dependent manner. In this study, we investigated the effects of TGF-{beta}1 (50 pM) or 10% newborn bovine serum (NBS) on modulation of paracellular passive ion conductance and its contribution to total passive ion conductance across RAECM. On days 5–7 in culture, tight-junctional resistance (Rtj, k{Omega}cm2) of RAECM, cultured in minimally defined serum-free medium (MDSF) with or without TGF-{beta}1 or NBS, was estimated from the relationship between observed transmonolayer voltage and resistance after addition of gramicidin D to apical potassium isethionate Ringer solution under open-circuit conditions. NaCl Ringer solution bathed the basolateral side throughout the experimental period. Results showed that transmonolayer conductance (1/Rte) and tight-junctional conductance (1/Rtj) are 0.59 and 0.14 mS/cm2 for control monolayers in MDSF, 1.59 and 0.38 mS/cm2 for monolayers exposed to TGF-{beta}1, and 0.38 and 0.18 mS/cm2 for monolayers grown in the presence of NBS. The contributions to total transepithelial ion conductance by the paracellular pathway are estimated to be 23, 23, and 47% for control, TGF-{beta}1-exposed, and newborn bovine serum (NBS)-treated RAECM, respectively.

air-blood barrier; permselectivity; paracellular resistance; barrier properties


Address for reprint requests and other correspondence: K.-J. Kim, Rm. HMR-914, Dept. of Medicine, Univ. of Southern California, Keck School of Medicine, 2011 Zonal Ave., Los Angeles, CA 90033 (E-mail: kjkim{at}usc.edu)




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