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Estimation of paracellular conductance of primary rat alveolar epithelial cell monolayers.
J Appl Physiol (1985) 2005; 98(1):138-43JA

Abstract

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 (R(te)) 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 (R(tj), kOmegacm(2)) 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/R(te)) and tight-junctional conductance (1/R(tj)) are 0.59 and 0.14 mS/cm(2) for control monolayers in MDSF, 1.59 and 0.38 mS/cm(2) for monolayers exposed to TGF-beta(1), and 0.38 and 0.18 mS/cm(2) 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.

Authors+Show Affiliations

Dept. of Medicine, University of Southern California, Keck School of Medicine, Rm. HMR-914, 2011 Zonal Avenue, Los Angeles, CA 90033, USA. kjkim@usc.eduNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

15273240

Citation

Kim, Kwang-Jin, et al. "Estimation of Paracellular Conductance of Primary Rat Alveolar Epithelial Cell Monolayers." Journal of Applied Physiology (Bethesda, Md. : 1985), vol. 98, no. 1, 2005, pp. 138-43.
Kim KJ, Borok Z, Ehrhardt C, et al. Estimation of paracellular conductance of primary rat alveolar epithelial cell monolayers. J Appl Physiol. 2005;98(1):138-43.
Kim, K. J., Borok, Z., Ehrhardt, C., Willis, B. C., Lehr, C. M., & Crandall, E. D. (2005). Estimation of paracellular conductance of primary rat alveolar epithelial cell monolayers. Journal of Applied Physiology (Bethesda, Md. : 1985), 98(1), pp. 138-43.
Kim KJ, et al. Estimation of Paracellular Conductance of Primary Rat Alveolar Epithelial Cell Monolayers. J Appl Physiol. 2005;98(1):138-43. PubMed PMID: 15273240.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Estimation of paracellular conductance of primary rat alveolar epithelial cell monolayers. AU - Kim,Kwang-Jin, AU - Borok,Zea, AU - Ehrhardt,Carsten, AU - Willis,Brigham C, AU - Lehr,Claus-Michael, AU - Crandall,Edward D, Y1 - 2004/07/23/ PY - 2004/7/27/pubmed PY - 2005/5/12/medline PY - 2004/7/27/entrez SP - 138 EP - 43 JF - Journal of applied physiology (Bethesda, Md. : 1985) JO - J. Appl. Physiol. VL - 98 IS - 1 N2 - 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 (R(te)) 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 (R(tj), kOmegacm(2)) 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/R(te)) and tight-junctional conductance (1/R(tj)) are 0.59 and 0.14 mS/cm(2) for control monolayers in MDSF, 1.59 and 0.38 mS/cm(2) for monolayers exposed to TGF-beta(1), and 0.38 and 0.18 mS/cm(2) 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. SN - 8750-7587 UR - https://www.unboundmedicine.com/medline/citation/15273240/Estimation_of_paracellular_conductance_of_primary_rat_alveolar_epithelial_cell_monolayers_ L2 - http://www.physiology.org/doi/full/10.1152/japplphysiol.00478.2004?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -