Novel aspects of the nervous system of Bonellia viridis (Echiura) revealed by the combination of immunohistochemistry, confocal laser-scanning microscopy and three-dimensional reconstruction

Abstract

The Echiura have been placed in close phylogenetic affinity to the Annelida on the basis of numerous homologous characters including the mode of development, the nearly identical formation of a trochophore larva, as well as the development and ultrastructure of chaetae and spermatozoa. Furthermore, phylogenetic analysis of elongation factor-1α gene sequences supports placement of the Echiura within the Annelida. Nevertheless, the Echiura are generally excluded from the Annelida due to their lack of segmentation. However, it must be considered that this lack could represent a secondary condition and that Echiura are derived from formerly segmented ancestors. In the present study, the combination of methods applied reveals several novel aspects of the central nervous system in developmental stages of Bonellia viridis. The most important of these is the metameric organization of the ventral nerve cord. Antibodies against different neurotransmitters label discrete repetitive units of perikarya in the ventral nerve cord. This organisation is additionally supported by the distribution of peripheral nerves as shown by labelling of neurotubules. These nerves are clearly paired and are evenly distributed, corresponding to the serial units of serotoninergic neurons. Different methods of computer-aided three-dimensional reconstruction display the precise spatial distribution of perikarya and peripheral nerves allowing the repetitive units to be discerned on the basis of relative size, position and number of labelled cells. The repetitive units in the nervous system of B. viridis correspond to segmental ganglia of various Annelida and are interpreted as an indication that Echiura are derived from formerly segmented ancestors, thus supporting the systematic inclusion of the Echiura within the Annelida.