Background: Arguments concerning the development of the pulmonary vein, and its relationship to the embryonic venous sinus (sinus venosus) have continued for well over a century. Recently, attention has again been focused on the origin of the pulmonary vein. It has been suggested that, whereas the pulmonary vein originates from the left atrium in humans, in all other vertebrates it originates from the venous sinus, with subsequent transfer to the left atrium. The nature of this transfer has not, however, been elucidated, although there is speculation that the pulmonary vein is ''pinched off'' from the left side of the embryonic venous sinus.Methods: We studied closely staged hearts of normal mouse embryos from a C57BL/6 x CBA cross days 10 and 11 of gestation (plug day ؍ day 1). Two series of embryos were collected and fixed in 2% glutaraldehyde, 1% formaldehyde, buffered with 0.05 M sodium cacodylate pH 7.4 (adjusted to 330 mOsm with NaCl). One series was wax embedded, serially sectioned, and stained with Masson's trichrome. The second series was subject to microdissection and scanning electron microscopy.Results: The atrial component of the heart tube is attached to the body of the embryo by reflections of the atrial myocardial wall. The attachment can be considered, from the outset, as the heart stalk, with the myocardialmesodermal connections forming a horseshoe of tissue that projects ventrally into the lumen of the atrium, surrounding a single evagination in the midline of the embryo. This heart stalk is cranial to the connections of the tributaries of the embryonic venous sinus and ventral to the foregut. When traced through its developmental stages, the evagination in the centre of the stalk, which we describe as the pulmonary pit, is seen to become the portal of entry for the developing pulmonary vein.Conclusions: The heart stalk, representing the area used by the pulmonary vein to gain access to the heart, and analogous to the dorsal mesocardium, is, from the outset, discrete from the area occupied by the orifices of the horns of the embryonic venous sinus. The pulmonary vein does not, in the mouse, develop from the tissues that form the walls of the tributaries of the systemic venous sinus. Comparisons with other studies suggest that early events in the development of the pulmonary vein are likely to be the same in all mammals, including humans. Anat.