Ultrastructural observations on spermiogenesis in the peanut worm,Themiste pyroides (Chamberlin, 1920) (Sipuncula; Sipunculidea)

Summary

The process of spermiogenesis and the ultrastructure of the spermatozoa in the peanut worm, Themiste pyroides, from the Sea of Japan were observed with electron microscopy (SEM and TEM). The testes are composed of groups of spermatogonia and are covered by peritoneal cells. Clusters of spermatocytes are released from the testes into the coelomic fluid. Connected by intercellular bridges, the spermatocytes within a given cluster develop asynchronously. Proacrosomal vesicles and a flagellum appear in spermatocytes. Spermatids in the clusters retain the intercellular connections. During spermiogenesis, the acrosomal vesicle, formed by coalescence of small proacrosomal vesicles in the basal part of the spermatid, migrates to the apical part of the cell to form a conical-shaped acrosome. The basal concavity lying above the nucleus is filled with subacrosomal substance. The midpiece contains four mitochondria, two centrioles, and some residual cytoplasm with dark glycogen-like granules. A peculiar annulus structure develops around the base of the flagellum. The distal centriole has a pericentriolar complex consisting of radially oriented elements. Before the spawning process, the spermatozoa are filtered throughout the ciliary nephrostomal funnel into the excretory sac of paired nephridia where they are stored for a short time. The sperm are released into the sea water via nephridiopores. Spermatozoa remaining in the coelomic fluid after spawning are resorbed by amoebocytes. This species from Vostok Bay is characterized by a prolonged spawning period from June to early October. The reproductive strategy of T. pyroides is discussed in comparison with that of Thysanocardia nigra, the latter having a unique pattern of packaging of the spermatozoa, resulting in the formation of spermatozeugmata, as a reproductive adaptation to the very short spawning period.     

星虫动物门活性物质及药理作用研究

本文简要介绍了星虫动物门的由来、分类和星虫的一般生理生态活性特征,并对星虫动物门的资源利用现状进行了分析,肯定了我国在星虫人工养殖、新药开发和利用其作为污染物监测方面所取得的成就.重点介绍了星虫动物门已经发现的营养活性物质、自身生理活性物质和药理活性物质以及星虫所具有的药理活性并对其未来研究进行了展望,比较了国内外学者对星虫研究的区别,为更好地保护和研究星虫提出了一些思路.     

Coelomocyte locomotion in the sipunculan Themiste petricola induced by exogenous and endogenous chemoattractants: role of a CD44-like antigen--HA interaction

Cell migration is a key event in the invertebrate immuno-defense system. Microbial products like lipopolysacharide (LPS) and formyl-methyl-leucyl-phenylalanine (fMLP) promote cell recruitment to sites of infection. In mammals, complement activation by factors such as zymosan induces C5a production, which influences leukocyte migration. The endogenous factor hyaluronic acid (HA), an extracellular matrix component, also promotes cell migration through its receptor CD44. We evaluated whether coelomocytes from the sipunculan worm T. petricola migrated towards LPS, fMLP, or zymosan treated plasma (ZTP) and if HA was involved in coelomocyte migration and adhesion. We also evaluated if antibodies specific for mouse HA receptor CD44 inhibited any of the effects induced by HA. Using microchemotaxis chambers we found that coelomocytes migrated towards exogenously and endogenously derived chemoattractants. We also observed that HA was a potent chemotactic signal and that coelomocytes adhered strongly to plates coated with LMW-HA but not with HMW-HA. In addition we found that these HA mediated effects were blocked by the monoclonal antibody IM7 directed to mouse CD44, suggesting that a CD44-like cross-reactive antigen might play a role in HA mediated coelomocyte locomotion.     

Fine structure of the pharyngeal apparatus of the pelagosphera larva in Phascolosoma agassizii (Sipuncula) and its phylogenetic significance

Sipuncula is a small taxon of worm-like marine organisms of still uncertain phylogenetic position. Sipunculans are characterized by an unsegmented body composed of a trunk into which the anterior part, the introvert, can be withdrawn. The group has been placed at various positions within Metazoa; currently, it is either seen as sister group of a clade comprising Mollusca and Annelida or as sister to each of these. An in-group position in either Mollusca or Annelida has usually been precluded till now due to the lack of so-called annelid or molluscan “key-characters” such as segmentation and chaetae or the radula. In the development of certain taxa the trochophore stage is followed by a planktonic larva, the pelagosphera, which might exhibit phylogenetically important structures. Among these is the buccal organ, which has been considered homologous either to the ventral pharyngeal organ present in many sedentary polychaetes or to the radular apparatus of molluscs. In the present paper, the ventral pharynx of the pelagosphera larva of Phascolosoma agassizii is investigated by transmission electron microscopy. The pharynx comprises dorsolateral ciliary folds, a muscle bulb formed by transverse muscle fibres with large intercellular spaces, and an investing muscle. A tongue-like organ is lacking. These results show great structural correspondences to the ventral pharynx of polychaetes, especially to that of the flabelligerid Diplocirrus longisetosus. In contrast, there are no signs of structural similarities to the corresponding structures of molluscs. Thus evidence increases that Sipuncula are closely related to annelids; moreover, an in-group position of Sipuncula within Annelida, as suggested by recent molecular studies, is not precluded by the present data. Instead these studies find additional support. Hence the lack of segmentation and chitinous chaetae in Sipuncula would be a secondary rather than a primary situation, as has recently been shown for Echiura and Pogonophora.     

Reconstructing the phylogeny of the Sipuncula

Sipunculans are marine spiralian worms with possible close affinities to the Mollusca or Annelida. Currently 147 species, 17 genera, 6 families, 4 orders and 2 classes are recognized. In this paper we review sipunculan morphology, anatomy, paleontological data and historical affiliations. We have conducted cladistic analyses for two data sets to elucidate the phylogenetic relationships among sipunculan species. We first analyzed the relationships among the 45 species of Phascolosomatidea with representatives of the Sipunculidea as outgroups, using 35 morphological characters. The resulting consensus tree has low resolution and branch support is low for most branches. The second analysis was based on DNA sequence data from two nuclear ribosomal genes (18S rRNA and 28S rRNA) and one nuclear protein-coding gene, histone H3. Outgroups were chosen among representative spiralians. In a third analysis, the molecular data were combined with the morphological data. Data were analyzed using parsimony as the optimality criterion and branch support evaluated with jackknifing and Bremer support values. Branch support for outgroup relationships is low but the monophyly of the Sipuncula is well supported. Within Sipuncula, the monophyly of the two major groups, Phascolosomatidea and Sipunculidea is not confirmed. Of the currently recognized families, only Themistidae appears monophyletic. The Aspidosiphonidae, Phascolosomatidae and Golfingiidae would be monophyletic with some adjustments in their definition. The Sipunculidae is clearly polyphyletic, with Sipunculus nudus as the sister group to the remaining Sipuncula, Siphonosoma cumanense the sister group to a clade containing Siphonosoma vastumand the Phascolosomatidea, and Phascolopsis gouldi grouping within the Golfingiiformes, as suggested previously by some authors. Of the genera with multiple representatives, only Phascolosoma and Themiste are monophyletic as currently defined. We are aiming to expand our current dataset with more species in our molecular database and more detailed morphological studies.     

An examination of the Phascolosotna subgenera Antillesoma, Rueppellisoma and Satonus (Sipuncula)

Three of the subgenera in the sipunculan genus Phascolosoma are reviewed, based on examination of type material. The presumed difference between the subgenera Antillesoma and Rueppellisoma (number of retractor muscles) is shown to be invalid and the taxa are therefore synonymous. The 15 species are either considered junior synonyms of P. antillarum (eight), transferred to other species or genera (six), or considered incertae sedis (one). Phascolosoma antillarum is redescribed. Of the eight species in the subgenus Satonus, only P. pectinatum remains valid: the others are considered either to belong to other taxa (four) or to be incertae sedis (three).     

Phylogenetic position of Sipuncula derived from multi‐gene and phylogenomic data and its implication for the evolution of segmentation

The phylogenetic position of Sipuncula, a group of unsegmented marine worms, has been controversial for several decades: Especially based on morphological data, closer relationships to Mollusca or Annelida were among the most favoured hypotheses. Increasing amounts of molecular data in recent years have consistently placed Sipuncula either in close affinity to or even within Annelida, the segmented worms, and rejected a close relationship to Mollusca. Yet, it remained uncertain whether Sipuncula is the sister group of Annelida or an annelid subtaxon. Therefore, herein we gathered data for five nuclear genes, which have been rarely used regarding Annelida and Sipuncula, and combined these with data for six previously used genes to further elucidate the phylogenetic position of Sipuncula. We also compiled a data set for 78 ribosomal proteins from publicly available genomic data sets. These are the two largest data sets for annelids with more than 10 taxa to date. All analyses placed Sipuncula within Annelida. For the first time, topology tests significantly rejected the possibility that Sipuncula is sister to Annelida. Thus, our analyses revealed that Sipuncula had secondarily lost segmentation. Given that unsegmented Echiura is also an annelid subtaxon, segmentation, a key character of Annelida, is much more variable than previously thought. Yet, this conclusion does not support the hypothesis that the last common ancestor of Annelida, Arthropoda and Chordata was segmented, assuming several losses along the branches leading to them. As yet no traces of segmentation could be shown in taxa exhibiting serially organized organ systems such as certain Mollusca, while in Sipuncula and Echiura such traces could be demonstrated. An independent origin of segmentation in Annelida, Arthropoda and Chordata thus appears to be more plausible and parsimonious.     

Free-swimming Cellular Complexes in the Coelom of the Sipunculid Thysanocardia nigra Ikeda, 1904 (Sipuncula)

The ultrastructural characteristics of coelomic cell complexes in the coelomic fluid were investigated with the use of transmission electron microscopy on the example of Japanese sipunculid. In the sipunculid coelom, complexes consisting of several cells were found for the first time: the central glandular cell and the outer layer of podocytes. Peculiar cell complexes (urns), comprising by ciliary and granular cells, were described in Thysanocardia for the first time. It had been proposed that both types of coelomic cell complexes dissociated from extensive chloragogenic tissue clusters on the intestine surface of Th. nigra. The variety of cell complexes in the coelom of other sipunculid is discussed.     

Embryonic and larval development of the peanut worm Phascolosoma agassizii (Keferstein 1867) from the Sea of Japan (Sipuncula: Phascolosomatidea)

All stages of the embryonic and larval development of Phascolosoma agassizii from Peter the Great Bay (Sea of Japan) were studied and illustrated using light and electron microscopy. The eggs of P. agassizii have the form of an ellipsoid (long and short axes about 100 and 70?µm, respectively). Egg cleavage is typical, spiral, and unequal. Gastrulation occurs by epiboly. This species possesses two pelagic larval stages, a lecithotrophic trochophore and a planktotrophic pelagosphera. The transformation of trochophore into pelagosphera occurs 80–90?h after fertilization. After 120–180?h, the larva has developed all systems of organs characteristic of the pelagosphera and is capable of feeding. At day 10, pelagospheras can settle, for some time, on the aquarium bottom and move on a ciliated lip, collecting food with the aid of a buccal organ. In addition, the larvae periodically attach themselves to the aquarium bottom or to the surface film of the water by means of a terminal organ. The trunk of the larva elongates by enlargement of the region behind the dorsal anal opening, which is located almost in the middle of the trunk region in the 15-day old larva. In the laboratory, 1-month old larvae spend the greater part of time in the attached state. Being attached by a glandular terminal organ to the aquarium bottom, they characteristically bend the body, actively feeding on microalgae from the substratum surface. The differences in the development of P. agassizii in the isolated West-Pacific and East-Pacific populations are shown and discussed.