Anuran Heart Metamorphosis: Anatomical Support for Pulmonary Blood Separation in the Early Aquatic Phase

1. Background In both larval and adult anurans, blood separation and respiratory physiology have remained an enigma. While various blood separation mechanisms have been proposed, the same structure is seen as playing a key role: the conus arteriosus. However, previous findings on its internal structure are contradictory, depending on the specifics of the 2D imaging methods used by different authors. To resolve this problem, we used high-resolution X-ray microtomography of whole Bufo bufo specimens to acquire the first detailed 3D descriptions of this complex structure through metamorphosis. Results In early tadpoles two small valvular openings develop at the ventricular-conal junction, providing two paths separated by the septum coni and continuing into the aortic arches. Thus, structures to support segregated pulmonary circulation are fully developed well before the lungs appear. The external gills undergo partial resorption and retreat asymmetrically into a gill chamber formed by a hyoidal cover, leaving only a single opening on the left side, the opercular spout. Conclusions The timing of events in Bufo circulatory development does not track the changing modes of respiration used by the developing tadpole. In particular, a system capable of double circulation carries only oxygen-depleted blood for a significant portion of the tadpole stage.