An "ancient" complexity? Evolutionary morphology of the circulatory system in Xiphosura

Horseshoe crabs (Xiphosura) have been an object of zoological research for almost 200 years. Although some morphological work on the circulatory system has been done, the three-dimensional structure of this complex organ system has never been shown satisfactorily and some crucial questions remain unanswered. Here, the circulatory systems of juveniles of the horseshoe crab taxa Limulus polyphemus and Carcinoscorpius rotundicauda were investigated using a combination of an injection method and micro-computed tomography. Data were processed and 3D-visualized using reconstruction software. Furthermore, the heart was examined using scanning electron microscopy. Additionally, the histology of some structures was investigated via light microscopy and transmission electron microscopy. The results show the high degree of complexity of the arterial and lacunar systems of Xiphosura and provide insights into their three-dimensional structure and relationship to other organ systems such as the central nervous system. We show that the major lacunae, previously described as vessel-like - though indeed highly ramified - can clearly be distinguished from arteries in histological sections because they have no distinct walls. Similarities and differences between the xiphosuran species and arachnids are highlighted and possible phylogenetic implications and evolutionary scenarios discussed.