In order to trace tissue movements during septation of the embryonic truncus arteriosus into aortic and pulmonary cardiac outlets, the cephalic margin of the developing tubular heart of chick embryos was tattooed at Hamilton-Hamburger Stages 20–22 using diffusion micropipettes filled with 0.5% agarose and radioactive macromolecular precursors (tritiated thymidine, uridine, and leucine). Following further incubation for 2, 48, or 96 hours, the locations of such tatoos were determined by autoradiography of sectioned tissue and computer reconstruction of the developing outflow tract.

Two hours after tattooing, radiolabeled cells were clustered at the right distal margin of the myocardial tube, as intended. Two days later, during septation of the outflow tract into the two arterial streams, label was concentrated along the posterior margin of the myocardium, between the developing aortic and pulmonary valve anlagen to the embryo's right and left, respectively. Four days following tattooing, as truncal septation neared completion, remaining label was found primarily to the left of the aortic valve ring posterior to the pulmonary outlet. The movements of thymidine tattoos during septation were demonstrated in a series of 31 embryos, 14 fixed at 2 hours, 12 at 2 days, and 5 at 4 days following tattooing; similar results were seen in uridine and leucine labeled hearts.

The motion of such tattoos in the developing chick heart suggests that the left side of the definitive semilunar valve ring derives from the right distal margin of the primitive tubular heart and that normal morphogenesis of the great arterial streams involves both retraction and rotation of the embryonic truncus arteriosus.