There is little information on the regulation of surfactant secretion in mouse type II cells. We isolated type II cells from C57BL/6 and FVB mice, cultured them overnight and then examined their response to known surfactant secretagogues. Secretion of phosphatidylcholine, SP-B and SP-C was stimulated by terbutaline, 5'-N-ethylcarboxyamidoadenosine (NECA), ATP, UTP, TPA and ionomycin. Phosphatidylcholine secretion was increased approximately 2-fold by all agonists in both strains of mice. The response to terbutaline and NECA is the same as in rat type II cells whereas the response to ATP, UTP, TPA and ionomycin is considerably less. Secretion of SP-B and SP-C was increased 7-fold by terbutaline and 3-fold by ATP, effects similar to those in rat type II cells. The response to terbutaline was significantly decreased in type II cells from ₂-adrenergic receptor null mice. These data establish that briefly cultured type II cells provide a suitable model for investigation of surfactant secretion in normal and genetically altered mice.