Patrick Beckers

Myzostomida are putatively nested within the errant Annelida and exhibit a unique morphology. The latter fact might possibly be related to their long host‐dependent radiation. Hence, an incomplete segmentation, lack of prominent sensory structures in adults and a dorso‐ventrally flattened body are just some examples. Although numerous investigations of the nervous system exist for myzostomids, detailed ultrastructural as well as histological examinations of neuronal structures are lacking so far. Therefore, we investigate the nervous system of Myzostoma cirriferum Leuckart, 1836 using a comparative approach including paraffin histology, serial semi‐thin sections, immunohistochemistry and ultrastructural investigations. Our analyses reveal a lack of complexity within the anterior‐most neuronal condensation (herein called brain) of adult specimens. Hence, prominent tracts or glomeruli are absent, and a glial layer surrounding the brain or radial‐like glial cells are lacking. Nevertheless, the ultrastructure of the ventral nervous system is comparable to other Errantia. Therefore, our investigations hint towards a reduction of neuronal complexity in Myzostomida. Additionally, an ontogenetic simplification seems plausible, although further investigations are necessary to verify such a hypothesis. A simplification of neuronal structures due to a drastic change in lifestyle was so far mainly observed for basally branching annelid clades.

Eunicida is a taxon of marine annelids currently comprising the taxa Eunicidae, Onuphidae, Dorvilleidae, Oenonidae, Lumbrineridae, Histriobdellidae and Hartmaniella. Most representatives are highly mobile hunters sharing the presence of a sophisticated nervous system but differ in the number and shape of prostomial sensory organs (0–3 antennae; 0 or 2 palps; 0, 2 or 4 (+2) buccal lips; 0, 2 or 4 eyes; single-grooved or paired nuchal organs). This makes Eunicida an ideal model to study the following questions: Is the brain morphology affected by different specificities of prostomial sensory organs? Do similar numbers and shapes of prostomial sensory organs hint at close phylogenetic relationships among different eunicidan taxa? How can antennae, palps and buccal lips be differentiated? For the investigation of sensory organs and the nervous system, we performed immunohistochemistry, µCT, TEM, SEM, paraffin histology and semi-thin sectioning. Our results show that brain anatomy is mostly affected on a microanatomical level by sensory organs and that similar specificities of sensory organs support the latest phylogenetic relationships of Eunicida. Further, a reduction of antennae in Eunicida can be suggested and hypotheses about the presence of sensory organs in the stem species of Eunicida are made.

Chaetopteridae — the parchment worms — comprise a group of early branching annelids with a scarcely investigated neuroanatomy and neurogenesis. Due to their phylogenetic position in the annelid tree, studying them is nevertheless inevitable for our understanding of character evolution in segmented worms. Therefore, we investigated several adult und larval chaetopterids using a broad set of morphological methods — including serial azan-stained histological sections as well as ultrastructural and immunohistochemical approaches. Our investigations shows that the chaetopterid nervous system consists of a medullary and intraepidermal anterior brain without major commissures and only one neuron type. Nuchal organs and complex cup-shaped eyes are absent in adult specimens. The developmental investigations reveal an antero-posterior origin of the larval nervous system, which is in line with previous investigations and supports this character as being plesiomorphic at least for Annelida. Furthermore, the reduction of neuronal complexity during ontogenesis hints towards the necessity of developmental examinations to understand the evolutionary scenarios behind nervous system diversity not only in annelid taxa. Our detailed investigations will help to deepen our knowledge in terms of annelid character evolution and will build up a basis for further detailed examinations dealing with this fascinating group of segmented worms.

Unfortunately, the captions of figures were incorrectly published in original publication. The corrected version updated here.

Mushroom bodies are known from annelids and arthropods and were formerly assumed to argue for a close relationship of these two taxa. Since molecular phylogenies univocally show that both taxa belong to two different clades in the bilaterian tree, similarity must either result from convergent evolution or from transformation of an ancestral mushroom body. Any morphological differences in the ultrastructure and composition of mushroom bodies could thus indicate convergent evolution that results from similar functional constraints. We here study the ultrastructure of the mushroom bodies, the glomerular neuropil, glia-cells and the general anatomy of the nervous system in Sthenelais boa. The neuropil of the mushroom bodies is composed of densely packed, small diameter neurites that lack individual or clusterwise glia enwrapping. Neurites of other regions of the brain are much more prominent, are enwrapped by glia-cell processes and thus can be discriminated from the neuropil of the mushroom bodies. The same applies to the respective neuronal somata. The glomerular neuropil of insects and annelids is a region of higher synaptic activity that result in a spheroid appearance of these structures. However, while these structures are sharply delimited from the surrounding neuropil of the brain by glia enwrapping in insects, this is not the case in Sthenelais boa. Although superficially similar, there are anatomical differences in the arrangement of glia-cells in the mushroom bodies and the glomerular neuropil between insects and annelids. Hence, we suppose that the observed differences rather evolved convergently to solve similar functional constrains than by transforming an ancestral mushroom body design.

Classical histological stained sections have the disadvantage that fine structures, like individual neurites, or specific macromolecules, like neurotransmitters cannot be visualized. Due to its highly specific staining of only one target molecule within the cell, the visualization of delicate structures, which would be superimposed by other tissue layers in classical Azan staining, is possible with immunohistochemistry. However, using immunohistological methods not all tissues of a specimen can be visualized at once. In contrast, density specific stains like Azan allow for a whole staining of the tissues. We provide a step by step protocol of how to combine immunohistochemistry and Azan staining in the same serial paraffin sections. The combination of both methods allows for a highly detailed investigation of structures of interest. The spatial detection of the previous, to Azan staining, gained antibody-labeled signal allows for a much better understanding of animal organ systems. By using serial sections, it is possible to create an aligned image stack that is both Azan stained and also antibody-labeled. Thus enabling a correlative approach that bridges traditional histology with immunohistochemistry in animal morphology.

The annelid anterior central nervous system is often described to consist of a dorsal prostomial brain, consisting of several commissures and connected to the ventral ganglionic nerve cord via circumesophageal connectives. In the light of current molecular phylogenies, our assumptions on the primary design of the nervous system in Annelida has to be reconsidered. For that purpose we provide a detailed investigation of the adult nervous system of Magelonidae – a putatively basally branching annelid family - and studied early stages of the development of the latter. Our comparative investigation using an integrative morphological approach shows that the nervous system of Magelonidae is located inside the epidermis. The brain is composed of an anterior compact neuropil and posteriorly encircles the prostomial coelomic cavities. From the brain two lateral medullary cords branch off which fuse caudally. Prominent brain structures such as nuchal organs, ganglia or mushroom bodies are absent and the entire nervous system is medullary. Our investigations also contradict previous investigations and present an updated view on established assumptions and descriptions. The comprehensive dataset presented herein enables a detailed investigation of the magelonid anterior central nervous system for the first time. The data reveal that early in annelid evolution complexity of brains and anterior sensory structures rises. Polymorphic neurons in clusters and distinct brain parts, as well as lateral organs - all of which are not present in outgroup taxa and in the putative magelonid sister group Oweniidae - already evolved in Magelonidae. Commissures inside the brain, ganglia and nuchal organs, however, most likely evolved in the stem lineage of Amphinomidae + Sipuncula and Pleistoannelida (Errantia+ Sedentaria). The investigation demonstrates the necessity to continuously question established descriptions and interpretations of earlier publications and the need for transparent datasets. Our results also hint towards a stronger inclusion of larval morphology and developmental investigations in order to understand adult morphological features, not only in Annelida.

Recent phylogenomic analyses congruently reveal a basal clade which consists of Oweniidae and Mageloniidae as sister group to the remaining Annelida. These results indicate that the last common ancestor of Annelida was a tube-dwelling organism. They also challenge traditional evolutionary hypotheses of different organ systems, among them the nervous system. In textbooks the central nervous system is described as consisting of a ganglionic ventral nervous system and a dorsally located brain with different tracts that connect certain parts of the brain to each other. Only limited information on the fine structure, however, is available for Oweniidae, which constitute the sister group (possibly together with Magelonidae) to all remaining annelids. The brain of Oweniidae is ring- shaped and basiepidermal. Ganglia, higher brain centers or complex sensory organs do not exist; instead the central nervous system is medullary. Posterior to the brain the ventral medullary cord arises directly from the ventral region of the brain in Myriowenia sp. while in Owenia fusiformis two medullary cords arise perpendicular to the brain ring, extend caudally and fuse posterior. The central nervous system is composed of a central neuropil and surrounding somata of the neurons. According to ultrastructural and histological data only one type of neuron is present in the central nervous system. The central nervous system of Oweniidae is the simplest in terms of enlargement of the dorsal part of the brain and neuron distribution found among Annelida. Our investigation suggests that neither ganglia nor commissures inside the brain neuropil or clusters of polymorphic neurons were present in the annelid stem species. These structures evolved later within Annelida, most likely in the stem lineage of Amphinomidae, Sipuncula and Pleistoannelida. Palps were supposedly present in the last common ancestor of annelids and innervated by two nerves originating in the dorsal part of the brain. A broader comparison with species of each major spiralian clade shows the medullary nervous system to be a common feature and thus possibly representing the ancestral state of the spiralian nervous system. Moreover, ganglia and clusters of polymorphic neurons seemingly evolved independently in the compared taxa of Spiralia and Annelida.