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1 Departments of Medicine, Physiology and Biophysics and Molecular Pharmacology and Toxicology, University of Southern California, Los Angeles, CA, USA; Department of Biomedical Engineering, University of Southern California, Los Angeles, CA, USA; Will Rogers Institute Pulmonary Research Center, University of Southern California, Los Angeles, CA, USA
2 Departments of Medicine, Physiology and Biophysics and Molecular Pharmacology and Toxicology, University of Southern California, Los Angeles, CA, USA; Department of Biochemistry and Molecular Biology, University of Southern California, Los Angeles, CA, USA; Department of Biopharmaceutics and Pharmaceutical Technology, Saarland University, Saarbruecken, Germany
3 Department of Biopharmaceutics and Pharmaceutical Technology, Saarland University, Saarbruecken, Germany
4 Department of Anesthesiology Critical Care Medicine, Childrens Hospital-USC, Los Angeles, CA, USA
5 Departments of Medicine, Physiology and Biophysics and Molecular Pharmacology and Toxicology, University of Southern California, Los Angeles, CA, USA; Department of Pathology, University of Southern California, Los Angeles, CA, USA; Will Rogers Institute Pulmonary Research Center, University of Southern California, Los Angeles, CA, USA
* To whom correspondence should be addressed. E-mail: kjkim{at}usc.edu.
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 morphologic and phenotypic characteristics of type I pneumocytes in vivo. We recently reported that transforming growth factor-beta1 (TGF-
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-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
cm2) of RAECM, cultured in minimally defined serum-free medium (MDSF) +/- TGF-1 or NBS, was estimated from the relationship between observed transmonolayer voltage (Vte) and resistance (Rte) following addition of gramicidin D to apical K-isethionate Ringer's solution under open-circuit conditions. NaCl Ringer's solution bathed the basolateral side throughout the experimental period. Results showed that transmonolayer conductance (Gte = 1/Rte) and tight junctional conductance (Gtj = 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-1, and 0.38 and 0.18 mS/cm2 for monolayers grown in the presence of NBS. The contribution to total transepithelial ion conductance by the paracellular pathway is estimated to be 23%, 23%, and 47% for control, TGF-1-exposed, and NBS-treated RAECM, respectively.
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