PURPOSE: To elucidate and compare the activity and mechanism of albumin uptake in primary cultured alveolar type II and type I-like epithelial cells. MATERIALS AND METHODS: Type II epithelial cells isolated from rat lungs were cultured for 2 days at 5 x 10(6) cells/35-mm dish or for 6 days at 2 x 10(6) cells/35-mm dish. The mRNA expression of marker genes and FITC-albumin uptake were examined. RESULTS: The cells cultured for 2 days exhibited cuboidal type II epithelial morphology with lamellar bodies inside the cells, while the cells cultured for 6 days exhibited squamous type I epithelial morphology. These morphological characteristics were consistent with the changes in mRNA expression pattern of marker genes. FITC-albumin uptake in both cells was temperature-dependent and was inhibited by metabolic inhibitors and bafilomycin A1. The rate of uptake was much higher in type II cells than type I-like cells. In both cells, FITC-albumin uptake was inhibited by clathrin mediated-endocytosis inhibitors, but not by caveolae mediated-endocytosis inhibitors. CONCLUSIONS: These findings indicate that albumin in alveolar lining fluid is internalized into type II and type I epithelial cells via clathrin-mediated endocytosis, and the rate of albumin uptake is higher in type II cells than type I cells.
PURPOSE: To elucidate and compare the activity and mechanism of albumin uptake in primary cultured alveolar type II and type I-like epithelial cells. MATERIALS AND METHODS: Type II epithelial cells isolated from rat lungs were cultured for 2 days at 5 x 10(6) cells/35-mm dish or for 6 days at 2 x 10(6) cells/35-mm dish. The mRNA expression of marker genes and FITC-albumin uptake were examined. RESULTS: The cells cultured for 2 days exhibited cuboidal type II epithelial morphology with lamellar bodies inside the cells, while the cells cultured for 6 days exhibited squamous type I epithelial morphology. These morphological characteristics were consistent with the changes in mRNA expression pattern of marker genes. FITC-albumin uptake in both cells was temperature-dependent and was inhibited by metabolic inhibitors and bafilomycin A1. The rate of uptake was much higher in type II cells than type I-like cells. In both cells, FITC-albumin uptake was inhibited by clathrin mediated-endocytosis inhibitors, but not by caveolae mediated-endocytosis inhibitors. CONCLUSIONS: These findings indicate that albumin in alveolar lining fluid is internalized into type II and type I epithelial cells via clathrin-mediated endocytosis, and the rate of albumin uptake is higher in type II cells than type I cells.
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