Christiane Todt

Phylogenomics reveals deep molluscan relationships

Evolutionary relationships among the eight major lineages of Mollusca have remained unresolved despite their diversity and importance. Previous investigations of molluscan phylogeny, based primarily on nuclear ribosomal gene sequences or morphological data, have been unsuccessful at elucidating these relationships. Recently, phylogenomic studies using dozens to hundreds of genes have greatly improved our understanding of deep animal relationships. However, limited genomic resources spanning molluscan diversity has prevented use of a phylogenomic approach. Here we use transcriptome and genome data from all major lineages (except Monoplacophora) and recover a well-supported topology for Mollusca. Our results strongly support the Aculifera hypothesis placing Polyplacophora (chitons) in a clade with a monophyletic Aplacophora (worm-like molluscs). Additionally, within Conchifera, a sister-taxon relationship between Gastropoda and Bivalvia is supported. This grouping has received little consideration and contains most (>95%) molluscan species. Thus we propose the node-based name Pleistomollusca. In light of these results, we examined the evolution of morphological characters and found support for advanced cephalization and shells as possibly having multiple origins within Mollusca.

The fauna of hydrothermal vents on the Mohn Ridge (North Atlantic)

Abstract The macrofauna of the newly discovered hydrothermal vent field on the Mohn Ridge at 71°N was investigated. Samples were collected during the cruise BIODEEP 2006 using the ROV ‘Bathysaurus’. A total of 180 species-level taxa were identified. The region contains very few vent-endemic species, but some species of Porifera, Crustacea and Mollusca may be vent-associated. Dense aggregations of motile non-vent species such as Heliometra glacialis and Gorgonocephalus eucnemis surrounded the vent area, but the area in general only held small numbers of sedentary animals. Calcareous sponges comprised an unusually high portion of the sponge species found and they constitute one of the first pioneers among the sessile invertebrates settling on these vents. Possible explanations for the structure of the fauna in the region are discussed.

Aplacophoran Mollusks Evolved from Ancestors with Polyplacophoran-like Features

Summary Mollusca is an animal phylum with vast morphological diversity and includes worm-shaped aplacophorans, snails, bivalves, and the complex cephalopods [1]. The interrelationships of these class-level taxa are still contentious [2, 3], but recent phylogenomic analyses suggest a dichotomy at the base of Mollusca, resulting in a monophyletic Aculifera (comprising the shell-less, sclerite-bearing aplacophorans and the eight-shelled polyplacophorans) and Conchifera (all other, primarily univalved groups) [4, 5]. The Aculifera concept has recently gained support via description of the fossil Kulindroplax, which shows both aplacophoran- and polyplacophoran-like features and suggests that the aplacophorans originated from a shelled ancestor [6], but the overall morphology of the last common aculiferan ancestor remains obscure. Here we show that larvae of the aplacophoran Wirenia argentea have several sets of muscles previously known only from polyplacophoran mollusks. Most of these are lost during metamorphosis, and we interpret them as ontogenetic remnants of an ancestor with a complex, polyplacophoran-like musculature. Moreover, we find that the first seven pairs of dorsoventral muscles develop synchronously in Wirenia, similar to juvenile polyplacophorans [7], which supports the conclusions based on the seven-shelled Kulindroplax. Accordingly, we argue that the simple body plan of recent aplacophorans is the result of simplification and does not represent a basal molluscan condition.

Phylogenomics of Lophotrochozoa with consideration of systematic error

&NA; Phylogenomic studies have improved understanding of deep metazoan phylogeny and show promise for resolving incongruences among analyses based on limited numbers of loci. One region of the animal tree that has been especially difficult to resolve, even with phylogenomic approaches, is relationships within Lophotrochozoa (the animal clade that includes molluscs, annelids, and flatworms among others). Lack of resolution in phylogenomic analyses could be due to insufficient phylogenetic signal, limitations in taxon and/or gene sampling, or systematic error. Here, we investigated why lophotrochozoan phylogeny has been such a difficult question to answer by identifying and reducing sources of systematic error. We supplemented existing data with 32 new transcriptomes spanning the diversity of Lophotrochozoa and constructed a new set of Lophotrochozoa‐specific core orthologs. Of these, 638 orthologous groups (OGs) passed strict screening for paralogy using a tree‐based approach. In order to reduce possible sources of systematic error, we calculated branch‐length heterogeneity, evolutionary rate, percent missing data, compositional bias, and saturation for each OG and analyzed increasingly stricter subsets of only the most stringent (best) OGs for these five variables. Principal component analysis of the values for each factor examined for each OG revealed that compositional heterogeneity and average patristic distance contributed most to the variance observed along the first principal component while branch‐length heterogeneity and, to a lesser extent, saturation contributed most to the variance observed along the second. Missing data did not strongly contribute to either. Additional sensitivity analyses examined effects of removing taxa with heterogeneous branch lengths, large amounts of missing data, and compositional heterogeneity. Although our analyses do not unambiguously resolve lophotrochozoan phylogeny, we advance the field by reducing the list of viable hypotheses. Moreover, our systematic approach for dissection of phylogenomic data can be applied to explore sources of incongruence and poor support in any phylogenomic data set. [Annelida; Brachiopoda; Bryozoa; Entoprocta; Mollusca; Nemertea; Phoronida; Platyzoa; Polyzoa; Spiralia; Trochozoa.]

Of tests, trochs, shells, and spicules: Development of the basal mollusk Wirenia argentea (Solenogastres) and its bearing on the evolution of trochozoan larval key features

BackgroundThe phylogenetic status of the aplacophoran mollusk taxon Solenogastres (Neomeniomorpha) is controversially discussed. Some authors propose the clade to represent the most basal branch within Mollusca, while others claim aplacophoran mollusks (Solenogastres and Caudofoveata) to be derived. Larval characters are central in these discussions, specifically the larval test (calymma, apical cap), the ontogeny of the epidermal scleritome, and the proposed absence of larval protonephridia. To date, developmental data are available for five solenogaster species, but most reports are incomplete and need confirmation.ResultsWirenia argentea deposit small batches of uncleaved embryos that are tightly enclosed by a smooth and transparent egg hull. Cleavage is spiral and unequal. The ciliated larvae hatch about 45 hours after deposition and swim actively in the water column. Within 48-60 hours after hatching they become mushroom-shaped with a pronounced apical cap partly enclosing a posterior trunk. The cells covering the apical cap are large and cleavage arrested. On the apical cap there is a prominent prototrochal band of compound cilia and an apical ciliary tuft and the trunk bears a terminal ciliary band (telotroch). Obscured by the apical cap, a ciliary band originates in the stomodaeal pore and surrounds the trunk. As development is proceeding, the trunk elongates and becomes covered by cuticle with the exception of a ventral ciliary band, the future foot. The larvae have a pair of protonephridia. At 5 days after hatching they begin to settle and within the following 7-9 days the apical cap is gradually reduced. Scattered epidermal sclerites form under the cuticle. Wirenia argentea lack iterated groups of sclerites at any developmental stage. At 40 days after hatching, the postlarvae have a fully developed foregut, but the midgut and hindgut are not yet interconnected.ConclusionsSolenogastres develop via a trochophore-like lecitotrophic larva with a preoral apical cap that at least partly represents an enlarged prototrochal area. Homology of this larval type (pericalymma larva) to test cell larvae of other spiralian clades is doubtful. The ontogeny of W. argentea does not provide any evidence for a derived status of Solenogastres within Mollusca.

Comparative neuroanatomy of Caudofoveata, Solenogastres, Polyplacophora, and Scaphopoda (Mollusca) and its phylogenetic implications

The nervous system of invertebrates is considered to be a very conservative organ system and thus can be helpful to elucidate questions of phylogenetic relationships. Up to now, comparative neuroanatomical studies have been mainly focused on arthropods, where in-depth studies on major brain structures are abundant. In contrast, except for Gastropoda and Cephalopoda, the nervous system of representatives of the second largest phylum of invertebrates, the Mollusca, is as yet hardly investigated. We therefore initiated an immunohistochemical survey to contribute new neuroanatomical data for several molluscan taxa, especially the lesser known Caudofoveata, Solenogastres, Polyplacophora, and Scaphopoda, focusing on the cellular architecture and distribution of neurotransmitters in the brain. Antisera against the widespread neuroactive substances FMRFamide and serotonin were used to label subsets of neurons. Both antisera were additionally used in combination with acetylated α-tubulin and the nuclear marker DAPI. This enables us to describe the morphology of the nervous system at a fine resolution and to compare its cellular architecture between different species of one taxon, as well as between different taxa of mollusks. On the basis of these results, the nervous system of caudofoveates seems to be most highly derived within the so-called basal (non-conchiferan) mollusks, and a monophyly of a clade Aplacophora could not be confirmed. In general, the brain as well as the remaining nervous system of the molluscan taxa investigated shows a great variability, suggesting a deep time origin of the diversification of this prominent protostome clade.

Hemocyanin in mollusks—A molecular survey and new data on hemocyanin genes in Solenogastres and Caudofoveata☆

The most common respiratory protein of mollusks is the blue, copper-containing hemocyanin (van Holde and Miller, 1995). It is not bound to hemocytes but suspended in the hemolymph. Its molecular mass ranges from 3500 10 to 8000 10 Da (dalton) or even more (Herskovits, 1988). These differences in molecular weight are due to the fact that the basic decamers that constitute the barrel-shaped protein may aggregate to didecamers or multidecameric elongated particles (Herskovits, 1988). In cephalopods and chitons (Polyplacophora), there are exclusively decamers, whereas in protobranch bivalves and gastropods the predominantly observed aggregation state is didecamers (Herskovits, 1988; van Holde andMiller, 1995; Lieb andMarkl, 2004; Bergmann et al., 2006, 2007; Gatsogiannis et al., 2007). A typical hemocyanin monomer is composed of eight globular functional units (FU), which are arranged like pearls on a string and termed FU-a to FU-h. An exception are cephalopod hemocyanins that contain only seven FUs. According to molecular clock calculations, the single FUs evolved within the early Precambrian, thus they were present already before the extant molluscan classes derived (Lieb et al., 2000; Lieb and Markl, 2004). Further analyses of hemocyanin-genes from cephalopods, gastropods, a protobranch bivalve, and polyplacophorans (the latter is unpublished data) showed that all hemocyanin genes possess highly conserved phase 1 linker-introns, which separate the individual FU-exons from each other [(FU-a-intron-FU-b-intron-FU-c-intron-FU-d-intron...) Lieb et al., 2001; Altenhein et al., 2002; Bergmann et al., 2006, 2007]. Since these introns are present in all molluscan hemocyanin genes, they seem to be ancient and probably at first used to border a sin-

The complete mitochondrial genome of Scutopus ventrolineatus (Mollusca : Chaetodermomorpha) supports the Aculifera hypothesis

BackgroundWith more than 100000 living species, mollusks are the second most diverse metazoan phylum. The current taxonomic classification of mollusks recognizes eight classes (Neomeniomorpha, Chaetodermomorpha, Polyplacophora, Monoplacophora, Cephalopoda, Gastropoda, Bivalvia, and Scaphopoda) that exhibit very distinct body plans. In the past, phylogenetic relationships among mollusk classes have been contentious due to the lack of indisputable morphological synapomorphies. Fortunately, recent phylogenetic analyses based on multi-gene data sets are rendering promising results. In this regard, mitochondrial genomes have been widely used to reconstruct deep phylogenies. For mollusks, complete mitochondrial genomes are mostly available for gastropods, bivalves, and cephalopods, whereas other less-diverse lineages have few or none reported.ResultsThe complete DNA sequence (14662 bp) of the mitochondrial genome of the chaetodermomorph Scutopus ventrolineatus Salvini-Plawen, 1968 was determined. Compared with other mollusks, the relative position of protein-coding genes in the mitochondrial genome of S. ventrolineatus is very similar to those reported for Polyplacophora, Cephalopoda and early-diverging lineages of Bivalvia and Gastropoda (Vetigastropoda and Neritimorpha; but not Patellogastropoda). The reconstructed phylogenetic tree based on combined mitochondrial and nuclear sequence data recovered monophyletic Aplacophora, Aculifera, and Conchifera. Within the latter, Cephalopoda was the sister group of Gastropoda and Bivalvia + Scaphopoda.ConclusionsPhylogenetic analyses of mitochondrial sequences showed strong among-lineage rate heterogeneity that produced long-branch attraction biases. Removal of long branches (namely those of bivalves and patellogastropods) ameliorated but not fully resolved the problem. Best results in terms of statistical support were achieved when mitochondrial and nuclear sequence data were concatenated.

Development of the nervous system in Solenogastres (Mollusca) reveals putative ancestral spiralian features

BackgroundThe Solenogastres (or Neomeniomorpha) are a taxon of aplacophoran molluscs with contentious phylogenetic placement. Since available developmental data on non-conchiferan (that is, aculiferan) molluscs mainly stem from polyplacophorans, data on aplacophorans are needed to clarify evolutionary questions concerning the morphological features of the last common ancestor (LCA) of the Aculifera and the entire Mollusca. We therefore investigated the development of the nervous system in two solenogasters, Wirenia argentea and Gymnomenia pellucida, using immunocytochemistry and electron microscopy.ResultsNervous system formation starts simultaneously from the apical and abapical pole of the larva with the development of a few cells of the apical organ and a posterior neurogenic domain. A pair of neurite bundles grows out from both the neuropil of the apical organ and the posterior neurogenic domain. After their fusion in the region of the prototroch, which is innervated by an underlying serotonin-like immunoreactive (−LIR) plexus, the larva exhibits two longitudinal neurite bundles - the future lateral nerve cords. The apical organ in its fully developed state exhibits approximately 8 to 10 flask-shaped cells but no peripheral cells. The entire ventral nervous system, which includes a pair of longitudinal neurite bundles (the future ventral nerve cords) and a serotonin-LIR ventromedian nerve plexus, appears simultaneously and is established after the lateral nervous system. During metamorphosis the apical organ and the prototrochal nerve plexus are lost.ConclusionsThe development of the nervous system in early solenogaster larvae shows striking similarities to other spiralians, especially polychaetes, in exhibiting an apical organ with flask-shaped cells, a single pair of longitudinal neurite bundles, a serotonin-LIR innervation of the prototroch, and formation of these structures from an anterior and a posterior neurogenic domain. This provides evidence for an ancestral spiralian pattern of early nervous system development and a LCA of the Spiralia with a single pair of nerve cords. In later nervous system development, however, the annelids deviate from all other spiralians including solenogasters in forming a posterior growth zone, which initiates teloblastic growth. Since this mode of organogenesis is confined to annelids, we conclude that the LCA of both molluscs and spiralians was unsegmented.

The nervous system of the basal mollusk Wirenia argentea (Solenogastres): a study employing immunocytochemical and 3D reconstruction techniques

Abstract The morphology of the nervous system of Wirenia argentea (Solenogastres) is described based on histological and immunocytochemical analyses using fluorescent markers against the neurotransmitters serotonin and FMRFamide, as well as acetylated α-tubulin, in combination with confocal laser-scanning microscopy. The nervous system displays a distinct tetraneury with fused cerebral ganglia, large pedal ganglia, and longitudinal nerve cords connected by regularly spaced connectives and commissures resulting in an orthogon-like arrangement. There is no indication of metamery. FMRFamidergic immunoreactivity is present in most compartments of the nervous system, while serotonergic immunoreactivity appears to be restricted to the longitudinal nerve cords, the cerebro-pedal commissure and part of the cerebral ganglion. The buccal system shows immunoreactivity against none of the neurotransmitters.