Type IV Collagen Production by Rat Pulmonary Alveolar Epithelial Cells

Abstract Maintenance and repair of the pulmonary alveolar basement membrane are critical processes for preserving normal alveolar structure and function. To elucidate the mechanisms that control type IV collagen production by pulmonary alveolar epithelial cells, type II pneumocytes from rat lungs were isolated and maintained in tissue culture as monolayers. Using Northern blot analysis and metabolic labeling, we found that the cultured epithelial cells express type IV collagen mRNA and incorporate [3H]proline into type IV collagen. To determine the effects of phenotype on control of type IV collagen synthesis, we took advantage of the well-described observation that isolated type II cells lose their distinctive type II features when cultured on plastic in serum-containing medium for 7 days and assume an appearance more like that of type I epithelial cells. We found that [3H]proline incorporation into type IV collagen increased from day 1 to day 2 and thereafter decreased gradually up to day 7. Despite this decrease in [3H]proline incorporation, type IV collagen mRNA increased over the same period. If the loss of type II cell characteristics was prevented by culturing the cells in EHS matrix, a basement membrane-like extracellular matrix, there was little increase in relative abundance of type IV collagen mRNA as compared with cells cultured on plastic. We therefore conclude that type IV collagen production by isolated pulmonary alveolar epithelial cells is controlled, at least in part, by translational or post-translational mechanisms. Furthermore, the full type II cell phenotype is not required for expression of type IV collagen genes. Cells that have adopted a more type I-like appearance during time in culture continue to contain high levels of type IV collagen mRNA and secrete low but detectable levels of type IV collagen.