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Identification of Three Genes of Known Function Expressed by Alveolar Epithelial Type I Cells

Will Rogers Institute Pulmonary Research Center and Division of Pulmonary and Critical Care Medicine, University of Southern California, Los Angeles, California

Address correspondence to: Renli Qiao, M.D., Ph.D., Division of Pulmonary and Critical Care Medicine, University of Southern California, IRD 602, 2020 Zonal Avenue, Los Angeles, CA 90033. E-mail: rqiao{at}usc.edu

To identify genes of known function expressed by type I (AT1) cells, changes in gene expression during transdifferentiation of alveolar epithelial cells (AEC) in primary culture from type II (AT2) to type I–like cell phenotype were evaluated. Total RNA from AEC on Day 0 or Day 8 was hybridized to a rat microarray for screening. Eight upregulated genes on Day 8 were selected for further investigation. Northern analysis confirmed upregulation of three of these genes, PAI-1, P2X4, and P15^INK4B. The corresponding proteins were evaluated in cultured AEC and results correlated with expression in AT1 cells. In AEC monolayers, all three proteins increased between Day 1 and Day 8. In mixed populations of freshly isolated rat lung cells, concurrent labeling with the AT1 cell-specific antibody, VIIIB2, localized these proteins to AT1 cells. In whole lung, all three proteins were detected in alveolar epithelium in a location consistent with expression in AT1 cells. Identification of novel AT1 cell genes of known function suggests an active role for AT1 cells in alveolar homeostasis. Furthermore, expression of these gene products in AT1-like cells, in freshly isolated AT1 cells, and AT1 cells in whole lung indicates that AT1-like cells reflect many of the properties of AT1 cells in situ.

Abbreviations: alveolar epithelial cells, AEC • aquaporin-5, AQP5 • fluorescein isothiocyanate, FITC • immunofluorescence microscopy, IFM • phosphate-buffered saline, PBS • reverse transcriptase–polymerase chain reaction, RT-PCR • sodium dodecyl sulfate, SDS • saline sodium citrate, SSC

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