The Influence of Secretory Products of Morula Cells on Phagocytes of the Holothurian Eupentacta fraudatrix (Djakonov et Baranova, 1958) (Sclerodactylidae: Dendrochirotida)

Russian Journal of Marine Biology - The influence of humoral products of morula cells on apoptosis, the concentration of cytokinin-like compounds, and expression of cell-surface receptors specific...     

Dexamethasone Modulation of the Interaction between Two Types of Phagocytes of the Holothurian <Emphasis Type="Italic">Eupentacta fraudatrix</Emphasis> (Djakonov et Baranova, 1958) (Holothuroidea: Dendrochirotida)

This study examines the interaction between two types of phagocytes (P1 and P2) of the holothurian Eupentacta fraudatrix and its in vitro modulation by dexamethasone. Our results indicate that inhibition of apoptosis in P1 phagocytes by P2 phagocytes was accompanied by increased activities of antioxidant enzymes and reduced synthesis of interleukin-1α-like substances. We hypothesize that P1-phagocyte-related effects occurred in response to a high level of hydrogen peroxide produced by P2 phagocytes. The reduced anti-apoptotic effect of P2-phagocyte supernatant during prolonged incubation (24 h) was accompanied by a decline in defense reactions in P1 phagocytes due to depletion of antioxidant enzymes (catalase, glutathione reductase, and glutathione transferase). Inhibition of apoptotis in P1 phagocytes associated with upregulation of antioxidant enzyme defense in response to P2 phagocytes preincubated with dexamethasone (100 µM) indicates that P2 phagocytes affect P1 phagocytes via a ROS-associated mechanism. Thus, our data provide evidence that P1 and P2 phagocytes exhibit their maximum activity at different stages of the immune response, thus causing inhibition of activity in target cells during prolonged exposure. Dexamethasone enhances these effects.     

Structure of the Digestive Tube in the Holothurian Eupentacta fraudatrix (Holothuroidea: Dendrochirota)

In the holothurian Eupentacta fraudatrix,the gut wall exhibits trilaminar organization. It consists of an inner digestive epithelium, a middle layer of connective tissue, and an outer mesothelium (coelomic epithelium). The pharynx, esophagus, and stomach are lined with a cuticular epithelium composed of T-shaped cells. The lining epithelium of the intestine and cloaca lacks a cuticle and consists of columnar vesicular enterocytes. Mucocytes are also encountered in the digestive epithelium. The connective tissue layer is composed of a ground substance, which houses collagen fibers, amoebocytes, morula cells, and fibroblasts. The gut mesothelium is a pseudostratified epithelium, which is dominated by peritoneal and myoepithelial cells and also includes the perikarya and processes of the neurons of the hyponeural plexus and vacuolated cells.     

Post-autotomy regeneration of respiratory trees in the holothurian Apostichopus japonicus (Holothuroidea, Aspidochirotida)

Specialised respiratory organs, viz. the respiratory trees attached to the dorsal part of the cloaca, are present in most holothurians. These organs evolved within the class Holothuroidea and are absent in other echinoderms. Some holothurian species can regenerate their respiratory trees but others lack this ability. Respiratory trees therefore provide a model for investigating the origin and evolution of repair mechanisms in animals. We conducted a detailed morphological study of the regeneration of respiratory trees after their evisceration in the holothurian Apostichopus japonicus. Regeneration of the respiratory trees occurred rapidly and, on the 15th day after evisceration, their length reached 15–20 mm. Repair involved cells of the coelomic and luminal epithelia of the cloaca. Peritoneocytes and myoepithelial cells behaved differently during regeneration: the peritoneocytes kept their intercellular junctions and migrated as a united layer, whereas groups of myoepithelial cells disaggregated and migrated as individual cells. Although myoepithelial cells did not divide during regeneration, the peritoneocytes proliferated actively. The contractile system of the respiratory trees was assumed to develop during regeneration by the migration of myoepithelial cells from the coelomic epithelium of the cloaca. The luminal epithelium of the respiratory trees formed as a result of dedifferentiation, migration and transformation of cells of the cloaca lining. The mode of regeneration of holothurian respiratory trees is discussed. This work was funded by a grant from the Russian Foundation for Basic Research (project no. 08–04–00284) to I.Y.D. and by a grant from the Far Eastern Branch of the Russian Academy of Sciences and the Russian Foundation for Basic Research (project no. 09–04–98547) to T.T.G.     

Gamete ultrastructure in the arctic holothurian Molpadia borealis M. Sars, 1859 (Holothuroidea: Molpadiidae)

This study examines the gonad condition and gamete morphology of the Arctic deep-sea holothurian Molpadia borealis M. Sars, 1859 (Molpadiidae) collected from the Kara Sea in September 2006. The intensive process of gametogenesis observed in the holothurian gonads was close to completion, suggesting upcoming spawning. The sperm ultrastructure in M. borealis is similar to that in most Holothuroidea. This species has classic flagellated sperm, echinosperm, which are typical of animals with external insemination. The sperm head has a 2.5-??m spherical nucleus with a proximal acrosome consisting of a spherical acrosomal vesicle surrounded by periacrosomal material. A single circular mitochondrion located in the sperm mid-piece surrounds the proximal and distal centrioles, which are arranged at an obtuse angle to each other. The eggs of M. borealis are approximately 300 ??m in diameter. This suggests indirect development with a planktotrophic larva. This type of development of M. borealis is probably related to life in high latitudes in the deep sea.     

Spermatozoon Ultrastructure of Two Holothurian Species of the Genus Cucumaria (Dendrochirotida,Holothuroidea) from the Sea of Japan

The ultrastructure of spermatozoa of Cucumaria japonica and a congeneric morphologically similar deep-sea species was studied. The spermatozoa of both C. japonica and C. conicospermium are similar to those of other holothurians: the acrosome is composed of an acrosomal granule and periacrosomal material; the centrioles lie at an acute angle to one another; and the proximal centriole is connected to the nuclear envelope by a flagellar rootlet. The spermatozoa of C. japonica differ from those of C. conicospermium in the shape of the head and the dimensions and position of the acrosome. In C. japonica, the acrosome is completely embedded in the nuclear fossa and measures 0.7 m. In C. conicospermium, only one-third of the acrosome is embedded in the nuclear fossa; this acrosome measures 1.3 m. A correlation between the structure of the sperm acrosome and that of the egg envelope is discussed.     

The ultrastructure of the morula cells of Eupentacta quinquesemita (Echinodermata: Holothuroidea) and their role in the maintenance of the extracellular matrix

The ultrastructure of the morula cells of Eupentacta quinquesemita and the distribution of these cells in the dermal connective tissue are described. Morula cells are abundant in the dermis and appear to function in the maintenance of the extracellular matrix (ECM) as a source of ground substance material. The synthetic activity of these cells is described in detail. Morula cells are filled with large secretory vesicles containing three electrondense materials which are derived from rough endoplasmic reticulum and Golgi activity. The synthetic product of these cells contains glycosaminoglycans and is secreted into the ECM by degranulation. The ultrastructural and histochemical similarity of the degranulation product to the ECM ground substance suggests that they are comprised of the same material. Morula cells appear to function primarily in connective tissues where ground substance predominates. The cells often contain secretory vesicles at various stages of formation, all of which eventually mature and degranulate. The synthetic pathway of the morula cells appears to result ultimately in the complete disruption and death of the cells. The function of morula cells in the holothuroid ECM is discussed, and the synthetic activity of the cells is compared with that of other secretory cells.     

Identification and embryonic expression of Wnt2, Wnt4, Wnt5 and Wnt9 in the millipede Glomeris marginata (Myriapoda: Diplopoda)

The Wnt genes encode secreted glycoprotein ligands that are key players during animal development. Previous studies revealed the presence of 12 classes of Wnt genes in protostomes, although lineage specific losses of Wnt genes are common. So far, the gene expression profile of only two complete sets of arthropod Wnt genes has been studied; these are the Wnt genes of the fly Drosophila melanogaster and the beetle Tribolium castaneum. Insects, however, do not represent good models for the understanding of Wnt gene evolution because several Wnt genes have been lost in the lineage leading to the insects, or within the different orders of insects. Comparative gene expression data from non-insect arthropods are rare and restricted to a subset of Wnt genes.This study aims to fill this gap and describes four newly detected Wnt genes from the millipede Glomeris marginata (Myriapoda: Diplopoda). Together with previous studies, now 11 Glomeris Wnt genes have been isolated and their expression has been studied. The only predicted but hitherto undetected Wnt gene is Wnt10. The new data provide a platform for the comparison of Wnt gene expression patterns in arthropods and reveal conserved as well as diverged aspects of Wnt gene expression in Arthropoda. Prominent expression of Wnt4 in dorsal tissue implies a role in dorsal segmentation and suggests that Wnt4 may be the predicted substitute for the previously reported missing expression of wg/Wnt1 in dorsal tissue.     

New data on asexual reproduction,autotomy, and regeneration in holothurians of the Order Dendrochirotida

The features of asexual reproduction, autotomy, and regeneration in five species of holothurians from the Order Dendrochirotida which live in the Nha Trang Bay of the South China Sea were studied. In Colochirus robustus, the ability to perform fission was corroborated. It was shown that Cladolabes schmeltzii can redivide without completing its regeneration after a previous division. This process is similar to the fragmentation of other holothurians. The features of autotomy and regeneration in Colochirus quadrangularis, Ohshimella ehrenbergi and Massinium magnum were described for the first time.     

Maintaining the line of defense: regeneration of Cuvierian tubules in the sea cucumber Holothuria forskali (Echinodermata, Holothuroidea)

When irritated, individuals of the sea cucumber Holothuria forskali expel a few Cuvierian tubules which lengthen, instantly become sticky, and rapidly immobilize most organisms with which they come into contact. After expulsion, the lost tubules are readily regenerated. When only a few tubules have been expelled, there is often a latent period before the regeneration starts. In contrast, when many tubules have been expelled, the regenerative process starts immediately but proceeds in successive waves of 10 to 30 tubules that begin to regenerate at 10-day intervals. However, in all cases, the complete regeneration of a given tubule takes about 5 weeks and may be divided into three successive phases: an initial repair phase including the overall 48-h post-autotomy period, a true regenerative phase taking about 4 weeks to complete, and a growth phase of about one more week. Initial regeneration events occur by epimorphosis, cell proliferation being essential to the regenerative process, whereas late events occur mainly by morphallaxis, with migration of the newly differentiated cells. The mesothelium is the tissue layer in which cell proliferation is the most precocious and the most important, involving both peritoneocytes and undifferentiated cells (which seem to be dedifferentiated peritoneocytes). As regeneration proceeds, the percentage of undifferentiated cells regularly decreases in parallel with the differentiation of granular (adhesive-secreting) cells and myocytes. The myocytes then separate off from the mesothelium and migrate within the connective tissue layer. Three types of pseudopodial cells follow one another in the tubule connective tissue during regeneration. Type 1 cells have all the characteristics of echinoderm phagocytes and may have a fibroblastic function, cleaning the connective tissue compartment before new collagen synthesis starts. Type 2 cells are rather undifferentiated and divide actively. The presence of type 3 cells is closely associated with the appearance of collagen fibers, and it is suggested that they have a fibroblastic function. In the inner epithelium, cells also divide actively, but only those in which spherules have not yet differentiated in the basal intraconnective processes. It appears, therefore, that in the three tissue layers of the tubules, regeneration proceeds by cell dedifferentiation, then proliferation, and finally by differentiation. Cuvierian tubules thus constitute a very efficient defensive mechanism: their large number, sparing use, and particular regeneration dynamics make them an almost inexhaustible line of defense maintained at limited energy cost.     

Asexual reproduction, evisceration, and regeneration in holothurians (Holothuroidea) from Nha Trang Bay of the South China Sea

Peculiarities of asexual reproduction, evisceration, and regeneration were studied in 13 holothurian species of the orders Dendrochirotida and Aspidochirotida from Nha Trang Bay of the South China Sea. Asexual reproduction by fission has been described for the first time for Cladolabes schmeltzii. It has been shown for the first time that transected C. schmeltzii and Colochirus robustus successfully regenerate not only their anterior, but also their posterior structures. It was found that Pseudocolochirus violaceus was able to regenerate the anterior part of the body and to eject viscera through the anus. The modes of evisceration and regeneration of the gut have been elucidated for ten species of aspidochirotids. It has been shown that the intestine of Holothuria scabra forms from two anlagen rather than from a single one, as previously believed.     

Development of respiratory trees in the holothurian <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> (Aspidochirotida: Holothuroidea)

The development of respiratory trees in the holothurian Apostichopus japonicus has been studied using light and electron microscopy. Primordial respiratory trees emerge in 2–3-mm-long animals (2 months after fertilization). They arise as two independent outgrowths from the dorsal wall of the anterior part of the cloaca. Upon first emerging and throughout the course of development, the left respiratory tree is longer than the right one. A common base develops in 4-mm-long animals (2–3 months after fertilization). In yearlings, the left respiratory tree grows into gaps between the loops of the intestinal tube interlaced with intestinal hemal vessels. The developing coelomic and luminal epithelia have ultrastructural peculiarities. The luminal epithelium of respiratory trees has been shown for the first time to comprise nerve cells and their processes. Characteristic structural features of the epithelia are shown to begin developing as early as in 4-mm-long animals (2–3 months after fertilization). In yearlings, the respiratory trees demonstrate definitive structural patterns and are entirely functional.     

The effect of particle size frequency distribution of the substratum on the burrowing ability of Chiridota rigida (Semper) (Echinodermata: Holothuroidea)

Chiridota rigida (Semper) burrows easily into well‐sorted substrata, moving between particles in coarse substrata and pushing aside particles in fine substrata. Chiridota rigida does not burrow easily into poorly sorted substrata because spaces which can be penetrated are not available and because the particles cannot be moved as a result of increased stability of the substratum. In poorly sorted substrata, burrowing ability increases with increases in the proportion of fine particles. It would appear that the distribution and abundance of Chiridota rigida and other chiridotids would be affected by the effect of the substratum on their ability to burrow.     

Parallelisms in the evolution of sea cucumbers (Echinodermata: Holothuroidea)

The importance of taking into account parallelisms in the evolution of morphological characters is analyzed for the taxonomy of the class Holothuroidea. The establishment of the order Dactylochirotida and classification of the order Elasipodida serve as examples to illustrate insufficient appreciation of parallelisms in Holothuroidea. The following characters, evolving independently in different groups of sea cucumbers, are considered: a stout skeleton, reduction of the calcareous ring and the body wall sclerites; similarity of body shape; similarity in the shape of tentacles; reduction in the number of tentacles from 12 to 10 in different fam-ilies and subfamilies of the order Synaptida. Based on the analysis of morphological and molecular data, the family Deimatidae is transferred from the order Elasipodida to the order Aspidochirotida. It is hypothesized that the concave cup-shaped sclerites with three to five rays occurring in the family Laetmogonidae (order Elasipodida) are of paedomorphic origin and correspond to the early growth stages of the laetmogonid wheels; the concave cross-shaped sclerites of the families Elpidiidae and Psychropotidae may have originated from laetmogonid concave cup-shaped sclerites. Emended diagnosis of the order Elasipodida is proposed. The family Vaneyellidae previously synonymized by the author with the family Cucumariidae is reestablished, and its emended diagnosis is also proposed.     

The distribution and dissociation of cyclic adenosine 3':5'-monophosphate-dependent protein kinases in adipose, cardiac, and other tissues.

In crude extracts of adipose tissue the protein kinase dissociates slowly at 30 degrees into regulatory and catalytic subunits in the presence of 700 mug per ml of histone or 0.5 M NaCl. If the kinase is first dissociated by adding 10 muM adenosine 3':5'-monophosphate (cAMP), reassociation occurs instantaneously after removal of the cAMP by Sephadex G-25 chromatography. In contrast, in crude xtracts of heart, the protein kinase dissociates rapidly in the presence of 700 mug per ml of histone or 0.5 M NaCl and reassociates slowly after removal of cAMP. These differences are accounted for by the existence of two types of protein kinases in these tissues, referred to as types I and II. DEAE-cellulose chromatography of extracts of adipose tissue produces only one peak of cAMP-dependent protein kinase activity (type II) which elutes between 0.15 and 0.25 M NaCl. Similar chromatography of heart extracts resolves enzyme activity into two peaks; a type I enzyme which elutes between 0.05 and 0.1 M and predominates (greater than 75% of total activity), and a type II enzyme which elutes between 0.15 and 0.25 M NaCl. The dissociation properties of the types I and II enzymes from heart and adipose tissue are retained after partial purification by DEAE-cellulose and Sepharose 6B chromatography. Rechromatography of the separated peaks of the cardiac enzymes does not change the elution pattern. Sucrose density gradient centrifugation and gel filtration studies indicate that the molecular weights of these enzymes are very similar. The type II enzyme isolated by DEAE-cellulose chromatography of heart extracts resembles the adipose tissue enzyme, i.e. it undergoes slow dissociation at 30 degrees in the presence of histone or 0.5 M NaCl. The adipose tissue kinase and the heart type II kinase are not identical, however, since they do not elute at exactly the same point on DEAE-cellulose columns. A survey of several tissues indicates the presence of type I and II protein kinases similar to the enzymes in adipose tissue and heart as determined by DEAE-cellulose chromatography of crude extracts and by dissociation of the enzymes with histone. The presence of MgATP prevents dissociation of type I enzyme from heart by 0.5 M NaCl or histone. The profile of the enzyme on DEAE-cellulose, however, is not changed...     

The fine structure of the buccal tentacles of Holothuria forskali (Echinodermata,Holothuroidea)

Summary Ultrastructural study of the buccal tentacles of Holothuria forskali revealed that each tentacle bears numerous apical papillae. Each papilla consists of several differentiated sensory buds.The epidermis of the buds is composed of three cell types, i.e. mucus cells, ciliated cells, and glandular vesicular cells (GV cells). The GV cells have apical microvilli; they contain bundles of cross striated fibrillae associated with microtubules. Ciliated cells have a short non-motile cilium. Bud epidermal cells intimately contact an epineural nervous plate which is located slightly above the basement membrane of the epidermis. The epineural plate of each bud connects with the hyponeural nerve plexus of the tentacle. This nerve plexus consists of an axonic meshwork surrounded in places by sheath cells. The buccal tentacles have well-developed mesothelial muscles. Direct innervation of these muscles by the hyponeural nerve plexus was not seen.It is suggested that the buccal tentacles of H. forskali are sensory organs. They would recognize the organically richest areas of the sediment surface through the chemosensitive abilities of their apical buds. Tentacles presumably trap particles by wedging them between their buds and papillae.     

Fine structure of the spermatozoon of the sea cucumber,Leptosynapta clarki (Echinodermata: Holothuroidea)

Summary The spermatozoon of the holothurian Leptosynapta clarki has a small circular head measuring about 3.0 at the greatest diameter, a midpiece containing a single mitochondrion and a tail flagellum measuring between 35 and 45 in length. The acrosomal region contains a granule measuring 0.7 in diameter which consists of electron dense material arranged in concentric lamellae. Five concentric very electron dense lamellae alternate with areas of much less electron dense material in the central region of the granule. This granule rests in an anterior nuclear depression.The nucleus is circular in shape and contains one or two unbound vacuoles which frequently contain a fine granular material. Posteriorly the nucleus is bounded by a large mitochondrion and an occasional Golgi complex. The proximal centriole which contains a lateral arm of dense material lies in a deep fossa projecting into the nucleus. The distal centriole lies posteriorly in the mitochondrial mass and gives rise to nine satellite projections and their Y-shaped connective extensions.The tail contains the 9 + 2 tubule arrangement and tapers at its distal end.This investigation was supported by a National Research Council grant to F. S. Chia.     

Catalogue of neritic-benthonic echinoderms (Echinodermata: Crinoidea, Echinoidea, Holothuroidea) from the Cuban Archipelago

This is an updated taxonomic catalogue of neritic-benthonic Echinodermata (Crinoidea, Echinoidea, Holothuroidea) from Cuba, that includes locality, range, habitat and depth where each species is found. The species list is based on the collections of the Oceanology Institute; Havana, and the "Felipe Poey" Museum (Havana University) Specimens for those collections were captured by a variety of field techniques, narcotized with Cloral hydrate, fixed in formalin and stored in 70-80% alcohol. A total of 53 species were recorded (eight Crinoids, 22 Echinoids and 23 Holothuroids), in 37 genera, 19 families and 12 orders. Ocnus suspectus (Ludwig, 1874) and Phyllophorus (Urodemella) occidentalis (Ludwig, 1885) are new records for Cuban waters.