This study utilized electron microscopy and enzyme histochemistry to examine the morphological steps associated with the formation of a lumen in the end-buds of the embryonic rat submandibular gland. Lumen formation involved the development of junctional complexes by the central cells of the end-buds late on the 16th day of gestation. These junctional complexes established the apical domains of the presecretory cells, and cytofilament aggregates were observed beneath the presumptive luminal plasma membranes. This central area of the end-buds showed strong myosin ATPase activity, presumably associated with the cytofilaments. On the 17th day of gestation, small lumina appeared at the sites delineated by the junctional complexes. The myosin ATPase activity appeared to surround the newly opened lumina. By the 18th day of gestation, microvilli were seen projecting into the lumina, and secretory granules were often visible in the apical cytoplasm near the luminal plasma membranes of the early secretory cells. Myosin ATPase activity was greatly reduced once the lumina had formed.