Characterization of mouse alveolar epithelial cell monolayers
Year: 2009
Type: article
Abstract: We investigated the influence of extracellular matrix on transport properties of mouse alveolar epithelial cell (AEC) monolayers (MAECM) and transdifferentiation of isolated mouse alveolar epithelial type II (AT2) cells into an alveolar epithelial type I (AT1) cell-like phenotype. Primary mouse AT2 cells plated on laminin 5-coated polycarbonate filters formed monolayers with transepithelial resistance (R(T)) and equivalent short-circuit current (I(EQ)) of 1.8 kOmega.cm(2) and 5.3 microA/cm(2), respectively, after 8 days in culture. Amiloride (10 microM), ouabain (0.1 mM), and pimozide (10 microM) decreased MAECM I(EQ) to 40%, 10%, and 65% of its initial value, respectively. Sequential addition of pimozide and amiloride, in either order, revealed that their inhibitory effects are additive, suggesting that cyclic nucleotide-gated channels contribute to amiloride-insensitive active ion transport across MAECM. Ussing chamber measurements of unidirectional ion fluxes across MAECM under short-circuit conditions indicated that net absorption of Na(+) in the apical-to-basolateral direction is comparable to net ion flux calculated from the observed short-circuit current: 0.38 and 0.33 microeq.cm(-2).h(-1), respectively. Between days 1 and 9 in culture, AEC demonstrated increased expression of aquaporin-5 protein, an AT1 cell marker, and decreased expression of pro-surfactant protein-C protein, an AT2 cell marker, consistent with transition to an AT1 cell-like phenotype. These results demonstrate that AT1 cell-like MAECM grown on laminin 5-coated polycarbonate filters exhibit active and passive transport properties that likely reflect the properties of intact mouse alveolar epithelium. This mouse in vitro model will enhance the study of AEC derived from mutant strains of mice and help define important structure-function correlations.
Institution: University of Southern California
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Awards: Mechanisms in the Remodeling of Lung Structure, Role of FoxP2 in AEC (trans)differentiation, Molecular programming of AEC differentiation: role of forkhead factors, G2961101580, G362058062, Absorption mechanisms for peptide/protein drugs via lung, G4200889435, AT1 vs AT2 cells: differential roles in alveolar function and biology, Regulation of lung alveolar epithelial protein transport, Nanoparticle properties and alveolar epithelial barrier/transport functions, Environmental Exposures, Host Factors and Human Disease, Microtubule-based Transport in Lacrimal Gland Function
OpenAlex ID: W2016488925