Entactin expression by rat lung and rat alveolar epithelial cells

Although the composition of the subepithelial basement membrane of the alveolar septum has been studied in detail, there is relatively little information about which cells produce it. We examined intact rat lung and isolated rat alveolar type II cells for the expression of entactin, an integral basement membrane component that binds laminin and type IV collagen. By Northern analysis, late gestation and early neonatal rat lungs expressed high levels of entactin mRNA whereas lungs from adult animals had only minimal levels of entactin mRNA. These latter findings were confirmed by in situ hybridization, which showed prominent signal for entactin mRNA in cells in the alveolar walls of neonatal animals and no signal for entactin mRNA in the alveolar walls of lungs from adult animals. The entactin mRNA throughout the alveolar walls of neonatal animals was not limited to cells that expressed surfactant-associated protein C mRNA, a marker of alveolar type II cells. Freshly harvested adult alveolar type II cells and alveolar type II cells in culture for < 6 days expressed none to minimal entactin mRNA or protein. However, with longer periods in culture, both entactin mRNA and entactin protein synthesis were evident and progressively increased. In situ hybridization indicated that >60% of the alveolar epithelial cells expressed entactin mRNA with increasing time in culture. When cultured on Engelbreth-Holm-Swarm matrix, alveolar type II cells showed the same time course of entactin mRNA expression as cells cultured on plastic. Neonatal lung mesenchymal cells produced abundant entactin in culture, consistent with the likelihood that these cells are the principal source of entactin in alveolar walls in the developing lung. These results indicate that entactin production in the normal alveolar wall occurs primarily during lung development and that mesenchymal cells are probably the principal source of production. However, because adult alveolar epithelial cells synthesize entactin in culture, it is possible that alveolar epithelium contributes to the entactin in the alveolar subepithelial basement membrane.