The production of pulmonary surfactant protein D (SP-D) increases abruptly during late gestation, and the accumulation of this protein in lung tissue is increased following the exposure of fetal rats to glucocorticoids in utero. To examine the regulation of these events, we administered dexamethasone (Dex; 1 mg/kg/day intramuscularly for 3 days), or saline, to timed-pregnant rats and harvested the lungs on days 19 to 21 of gestation. Samples of pooled fetal lungs were analyzed for SP-D protein, mRNA, and gene transcription by immunoblot, Northern hybridization, and nuclear run-off transcription assays. Lungs from 19 day controls showed barely detectable levels of SP-D gene transcription and negligible accumulation of SP-D message. However, SP-D transcription and the accumulation of SP-D mRNA and protein were readily detected in lungs from 19 day Dex-treated rats. Dexamethasone also caused dose- and time-dependent increases in SP-D protein and mRNA accumulation in 19 day fetal lung explants. Immunohistochemistry of control 19 day lung using antibodies to rat SP-D showed only weak labeling of a small number of airway epithelial cells. By contrast, Dex-exposed rats showed strong staining of columnar and cuboidal epithelial cells lining airways and epithelial tubules and cuboidal cells lining primitive air sacs. In situ hybridization assays showed similar alterations in the number, intensity, and distribution of labeled epithelial cells in 19 day Dex-exposed lungs and demonstrated labeling of alveolar type II and nonciliated columnar cells in adult lung. These data indicate that the accelerated lung maturation accompanying glucocorticoid exposure in utero is associated with a precocious increase in SP-D gene transcription and protein production by pulmonary epithelial cells.