Spadella Cephaloptera Research Articles

Flow generation by the corona ciliata in Chaetognatha − quantification and implications for current functional hypotheses

The corona ciliata of Chaetognatha (arrow worms) is a circular or elliptical groove lined by a rim from which multiple lines of cilia emanate, located dorsally on the head and/or trunk. Mechanoreception, chemosensation, excretion, respiration, and support of reproduction have been suggested to be its main functions. Here we provide the first experimental evidence that the cilia produce significant water flow, and the first visualisation and quantification of this flow. In Spadella cephaloptera, water is accelerated toward the corona ciliata from dorsal and anterior of the body in a funnel-shaped pattern, and expelled laterally and caudally from the corona, with part of the water being recirculated. Maximal flow speeds were approximately 140μms−1 in adult specimens. Volumetric flow rate was Q=0.0026μls−1. The funnel-shaped directional flow can possibly enable directional chemosensation. The flow measurements demonstrate that the corona ciliata is well suited as a multifunctional organ.

Ribin-like proteins expression in the chaetognath Spadella cephaloptera

In chaetognaths, a marine phylum of approximately 120 species, paralogous ribosomal protein genes and two classes of both 18S and 28S rRNA gene sequences have been evidenced. Moreover, differential and specific expression of the rRNA genes has been demonstrated suggesting implications of regulatory mechanisms in the synthesis of the ribosome constituents. Results of in situ hybridization of ribin-like mRNAs in Spadella cephaloptera were compared with immunofluorescence localization of the corresponding protein. Previous studies showed that in rat Ribin protein is encoded on the antisense strand of the 28S rRNA gene and known as rRNA promoteur binding protein. In chaetognaths, expression of ribin-like gene(s) is restricted to oocytes similarly to both18S and 28S class II genes and to gut, whereas Ribin-like proteins have been identified in the nervous system, oocytes and gut. Chaetognath nuclear regions, homologous to ratribin mRNA have been sequenced; however, no complete open reading frame can be reached. Analysis of DNA databases reveals that deduced sequences of several animal nuclear sequences exhibit a region having a great level of conserved amino-acids with the COOH-part of the rat Ribin, suggesting a high selection pressure in this region. Moreover, analysis of EST libraries shows that Ribin-like genes are expressed in both animals and plants. In addition, in invertebrates, these transcripts are principally found during the first stages of development or in nervous tissues or in infected/stressed organisms. The selective expression and translation of Ribin-like genes added to their large evolutionary distribution suggest important physiological roles of the corresponding proteins. Key words: Ribin, chaetognath, nervous system, oocytes, gut.

High level of structural polymorphism driven by mobile elements in the Hox genomic region of the Chaetognath Spadella cephaloptera.

Little is known about the relationships between genome polymorphism, mobile element dynamics, and population size among animal populations. The chaetognath species Spadella cephaloptera offers a unique perspective to examine this issue because they display a high level of genetic polymorphism at the population level. Here, we have investigated in detail the extent of nucleotide and structural polymorphism in a region harboring Hox1 and several coding genes and presumptive functional elements. Sequencing of several bacterial artificial chromosome inserts representative of this nuclear region uncovered a high level of structural heterogeneity, which is mainly caused by the polymorphic insertion of a diversity of genetic mobile elements. By anchoring this variation through individual genotyping, we demonstrated that sequence diversity could be attributed to the allelic pool of a single population, which was confirmed by detection of extensive recombination within the genomic region studied. The high average level of nucleotide heterozygosity provides clues of selection in both coding and noncoding domains. This pattern stresses how selective processes remarkably cope with intense sequence turnover due to substitutions, mobile element insertions, and recombination to preserve the integrity of functional landscape. These findings suggest that genome polymorphism could provide pivotal information for future functional annotation of genomes.

Complete mitochondrial genome sequences of the three pelagic chaetognaths Sagitta nagae, Sagitta decipiens and Sagitta enflata

The complete nucleotide sequences of the mitochondrial genomes were determined for the three pelagic chaetognaths, Sagitta nagae, Sagitta decipiens, and Sagitta enflata. The mitochondrial genomes of these species which were 11,459, 11,121, and 12,631 bp in length, respectively, contained 14 genes (11 protein-coding genes, one transfer RNA gene, and two ribosomal RNA genes), and were found to have lost 23 genes that are present in the typical metazoan mitochondrial genome. The same mitochondrial genome contents have been reported from the benthic chaetognaths belonging to the family Spadellidae, Paraspadella gotoi and Spadella cephaloptera. Within the phylum Chaetognatha, Sagitta and Spadellidae are distantly related, suggesting that the gene loss occurred in the ancestral species of the phylum. The gene orders of the three Sagitta species are markedly different from those of the other non-Chaetognatha metazoans. In contrast to the region with frequent gene rearrangements, no gene rearrangements were observed in the gene cluster encoding COII–III, ND1–3, srRNA, and tRNA met. Within this conserved gene cluster, gene rearrangements were not observed in the three Sagitta species or between the Sagitta and Spadellidae species. The gene order of this cluster was also assumed to be the ancestral state of the phylum.

Fine structure of the ventral nerve centre and interspecific identification of individual neurons in the enigmatic Chaetognatha

The enigmatic arrow worms (Chaetognatha) are marine carnivores and among the most abundant planktonic organisms. Their phylogenetic position has been heavily debated for a long time. Most recent molecular studies still provide a diverging picture and suggest arrow worms to be some kind of basal protostomes. In an effort to understand the organization of the nervous system in this clade for a broad comparison with other Metazoa we analysed the ultrastructure of the ventral nerve centre in Spadella cephaloptera by transmission electron microscopy. We were able to identify six different types of neurons in the bilateral somata clusters by means of the cytoplasmic composition (regarding the structure of the neurite and soma including the shape and eu-/heterochromatin ratio within the nucleus) as well as the size and position of these neurons. Furthermore, our study provides new insights into the neuropil composition of the ventral nerve centre and several other fine structural features. Our second goal was to examine if individually identifiable neurons are present in the ventral nerve centres of four chaetognath species, Sagitta setosa, Sagitta enflata, Pterosagitta draco, and Spadella cephaloptera. For that purpose, we processed whole mount specimens of these species for immunolocalization of RFamide-related neuropeptides and analysed them with confocal laser-scanning microscopy. Our experiments provide evidence for the interspecific homology of individual neurons in the ventral nerve centres of these four chaetognath species suggesting that the potential to generate serially arranged neurons with individual identities is part of their ground pattern.

Transcriptome Analysis of ESTs from a Chaetognath Reveals a Deep-Branching Clade of Retrovirus-Like Retrotransposons

Chaetognaths constitute a small marine phylum exhibiting several characteristic which are highly unusual in animal genomes, including two classes of both rRNA and protein ribosomal genes. As in this phylum presence of retrovirus-like elements has never been documented, analysis of a published expressed sequence tag (EST) collection of the chaetognath Spadella cephaloptera has been made. Twelve sequences representing transcript sections of reverse transcriptase domain of active retrotransposons were isolated from~11,000 ESTs. Five of them are originated from Gypsy retrovirus-like elements, whereas the other are transcripts from a Bel-Pao LTR-retrotransposon, a Penelope-like element and LINE retrotransposons. Moreover, a part of a putative integrase has also been found. Phylogenetic analyses suggest a deep-branching clade of the retrovirus-like elements, which is in agreement with the probably Cambrian origin of the phylum. Moreover, retrotransposons have not been found in telomeric-like transcripts which are probably constituted by both vertebrate and arthropod canonical repeats.

Careful with understudied phyla: The case of chaetognath

BackgroundA recent study by Barthélémy et al. described a set of ribosomal protein (RP) genes extracted from a collection of expressed sequence tags (ESTs) of the chaetognath (arrow worm) Spadella cephaloptera. Three main conclusions were drawn in this paper. First, the authors stated that RP genes present paralogous copies, which have arisen through allopolyploidization. Second, they reported two alternate nucleotide stretches conserved within the 5' untranslated regions (UTR) of multiple ribosomal cDNAs and they suggested that these motifs are involved in the differential transcriptional regulation of paralogous RP genes. Third, they claimed that the phylogenetic position of chaetognaths could not be accurately inferred from a RP dataset because of the persistence of two problems: a long branch attraction (LBA) artefact and a compositional bias.ResultsWe reconsider here the results described in Barthélémy et al. and question the evidence on which they are based. We find that their evidence for paralogous copies relies on faulty PCR experiments since they attempted to amplify DNA fragments absent from the genomic template. Our PCR experiments proved that the conserved motifs in 5'UTRs that they targeted in their amplifications are added post-transcriptionally by a trans-splicing mechanism. Then, we showed that the lack of phylogenetic resolution observed by these authors is due to limited taxon sampling and not to LBA or to compositional bias. A ribosomal protein dataset thus fully supports the position of chaetognaths as sister group of all other protostomes. This reinterpretation demonstrates that the statements of Barthélémy et al. should be taken with caution because they rely on inaccurate evidence.ConclusionThe genomic study of an unconventional model organism is a meaningful approach to understand the evolution of animals. However, the previous study came to incorrect conclusions on the basis of experiments that omitted validation procedures.

Translational machinery of the chaetognath Spadella cephaloptera: a transcriptomic approach to the analysis of cytosolic ribosomal protein genes and their expression

BackgroundChaetognaths, or arrow worms, are small marine, bilaterally symmetrical metazoans. The objective of this study was to analyse ribosomal protein (RP) coding sequences from a published collection of expressed sequence tags (ESTs) from a chaetognath (Spadella cephaloptera) and to use them in phylogenetic studies.ResultsThis analysis has allowed us to determine the complete primary structures of 23 out of 32 RPs from the small ribosomal subunit (SSU) and 32 out of 47 RPs from the large ribosomal subunit (LSU). Ten proteins are partially determined and 14 proteins are missing. Phylogenetic analyses of concatenated RPs from six animals (chaetognath, echinoderm, mammalian, insect, mollusc and sponge) and one fungal taxa do not resolve the chaetognath phylogenetic position, although each mega-sequence comprises approximately 5,000 amino acid residues. This is probably due to the extremely biased base composition and to the high evolutionary rates in chaetognaths. However, the analysis of chaetognath RP genes revealed three unique features in the animal Kingdom. First, whereas generally in animals one RP appeared to have a single type of mRNA, two or more genes are generally transcribed for one RP type in chaetognath. Second, cDNAs with complete 5'-ends encoding a given protein sequence can be divided in two sub-groups according to a short region in their 5'-ends: two novel and highly conserved elements have been identified (5'-TAATTGAGTAGTTT-3' and 5'-TATTAAGTACTAC-3') which could correspond to different transcription factor binding sites on paralog RP genes. And, third, the overall number of deduced paralogous RPs is very high compared to those published for other animals.ConclusionThese results suggest that in chaetognaths the deleterious effects of the presence of paralogous RPs, such as apoptosis or cancer are avoided, and also that in each protein family, some of the members could have tissue-specific and extra-ribosomal functions. These results are congruent with the hypotheses of an allopolyploid origin of this phylum and of a ribosome heterogeneity.

The differential expression of ribosomal 18S RNA paralog genes from the chaetognath Spadella cephaloptera

Chaetognaths constitute a small marine phylum of approximately 120 species. Two classes of both 18S and 28S rRNA gene sequences have been evidenced in this phylum, even though significant intraindividual variation in the sequences of rRNA genes is unusual in animal genomes. These observations led to the hypothesis that this unusual genetic characteristic could play one or more physiological role(s). Using in situ hybridization on the frontal sections of the chaetognath Spadella cephaloptera, we found that the 18S Class I genes are expressed in the whole body, with a strong expression throughout the gut epithelium, whereas the expression of the 18S Class II genes is restricted to the oocytes. Our results could suggest that the paralog products of the 18S Class I genes are probably the “housekeeping” 18S rRNAs, whereas those of class II would only be essential in specific tissues. These results provide support for the idea that each type of 18S paralog is important for specific cellular functions and is under the control of selective factors.

Comparison of chaetognath mitochondrial genomes and phylogenetical implications

Two complete mitochondrial genomes (mtDNAs) of chaetognaths, Spadella cephaloptera and Paraspadella gotoi, have been recently published. These genomes are highly unusual. They are the smallest metazoan mtDNAs so far known; atp6 and atp8 genes are missing; lastly, our reanalysis has evidenced that, contrarily to what has been previously published for one sequence, both contain a unique transfer RNA (tRNA Met) evidencing that both have the same gene content. Indeed, even if the gene order seems very different, two gene blocks are conserved. In addition, comparison of gene arrangement suggests phylogenetical relationships between chaetognaths and some lophotrochozoa like annelids and molluscs.

Restricted expression of a median Hox gene in the central nervous system of chaetognaths

Hox genes encode a set of evolutionarily conserved transcription factors that regulate anterior-posterior patterning. Here we report the first developmental expression of a Hox gene from Chaetognatha. These metazoans have been shown recently to be part of the protostome group of bilaterians. We describe the analysis of the SceMed4 gene (a Spadella cephaloptera Median Hox gene) including its expression from late stages of egg development to 7 days after hatching. In all of these stages, SceMed4 is expressed in two lateral stripes in a restricted region of the developing ventral ganglion.

Wound healing and survival of beheaded chaetognaths

The observation in natural conditions of headless chaetognaths with healed scars and ripe gonads led us to reproduce this condition experimentally. Histological sections were made on specimens of Sagitta and Spadella at different times after beheading. The muscles contract and the gut closes the wound, so that by the end of the first two hours, the body wall and gut ‘basement membranes’ are in continuity, so restoring the integrity of the body. After two hours, the muscular layer of the body wall is reshaped and closes up the wound; the gut seals off, and a ‘clot’ is evident in the wound area. The benthic Spadella cephaloptera survives some 30 d or more after decapitation. Specimens carrying ripe ova were able to lay them, to mature and fill their seminal vesicles once or twice and to mate with normal mature specimens. Headless chaetognaths must take up nutrients through the integument. It appears that chaetognaths cannot really regenerate head or tail tip.

Morphology and ultrastructure of the gut in Spadella cephaloptera (chaetognatha).

Structural and ultrastructural studies on the gut of the chaetognath Spadella cephaloptera, as well as observations on the feeding behavior of specimens bred in the laboratory, were conducted. The gut displays four distinct zones: pharynx, esophagus, and intestine, to which are connected a pair of diverticula, and the rectum, differing in length, shape, and cellular composition. The intestine alone represents ∼90% of the gut length. Upon ingestion, food in the intestine is submitted to successive backward and forward peristaltic movements until digestion has ended. Ultrastructural observations have identified five distinct cell types from granule morphology and the presence or absence of cilia at the apex of the cells. Three of the types undoubtedly correspond to secretory cells. They are the (1) pharyngeal, (2) esophageal, and (3) light intestinal ciliated cells, which could be, respectively, implicated in (1) mucous, (2) enzyme, and (3) both mucous and enzyme secretions. The fourth type, which corresponds to dense intestinal ciliated cells, displays all the characteristics of cells specialized in the absorption of macromolecules and intracellular digestion. The products of this digestion could be temporarily stored inside dense granules before being utilized during vitellogenesis. Except for the presence of cilia, the fifth type, which is localized in the short rectum, represents a common polyhedral epithelial cell type. © 1996 Wiley-Liss, Inc.

Occurrence of Spadella cephaloptera During One Year at Ischia Island (Gulf of Naples)

Abstract. The life cycle of the benthic arrowworm, Spadella cephaloptera, which inhabits the Posidonia meadow at Ischia (Gulf of Naples, Italy), was examined by a one‐year systematic sampling survey and by laboratory culture. The seasonal fluctuations in abundance and the reproductive characteristics indicate that there were two generations of S. cephaloptera in one year; these generations have a different life span. The habitat of S. cephaloptera depends on the growth of the Posidonia meadow; the influence of different factors on the S. cephaloptera population, especially of the Posidonia vagile fauna, is discussed.

Prey detection in the Chaetognatha: Response to a vibrating probe and experimental determination of attack distance in large aquaria1

The benthic Spadella schizoptera and the planktonic Sagitta hispida both attacked a fine vibrating glass probe over a broad range of frequencies after being starved for 1–3 days. The response curve for S. schizoptera had a peak at 30 Hz and was similar in shape but far broader than that found by prior workers for Spadella cephaloptera. Sagitta hispida responded differently, with a peak response at 150 Hz and no apparent preference among lower frequencies. It also had a strong (37%) response to nonvibrating lateral sweeps.Neither species would attack the probe at a distance >3 mm. This extremely short attack distance was confirmed, for S. hispida, in a 50‐liter aquarium by a series of feeding experiments, the results from which were applied to a simple model. Sagitta hispida starved for 1 day searched 2,667 mm3 min−1 and had an attack distance of 0.88 mm. These experimental data were used to determine the ability of S. hispida to sustain itself nutritionally in its natural environment.

Prey detection by Chaetognatha via a vibration sense

Chaetognaths (arrow worms) have two types of tufts along the body, not previously distinguished. The bristles seen in life are non-sensory processes of epithelial cells. The ciliary tufts visible in sections are groups of sensory neurons each of which bears a non-motile cilium at the tip of its dendrite. The high sensitivity to vibrations set up by a nearby oscillating source in the dark is attributed to the non-motile cilia. The example analysed, Spadella cephaloptera , makes an accurate feeding movement towards any source vibrating at 9 to 20 c/s with an amplitude of 100 to 500 μm at a distance of 1 to 3 mm. Other vibrations are ignored or cause an escape response.

Habits, Structure, and Development of Spadella Cephaloptera

Habits, Structure, and Development of Spadella Cephaloptera