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Close relationship between Asterina and Solasteridae (Asteroidea) supported by both nuclear and mitochondrial gene molecular phylogenies.

Phylogenetic relationships among asteroids remain to be extremely controversial in spite of many morphological and molecular studies have been applied to this issue. In the present study, especially focusing on resolving the relationship of Asterina and Solasteridae, we reconstructed the molecular phylogenetic tree of asteroids using nuclear 18S rDNA. A close relationship between Asterina and Solasteridae, which has been suggested from analyses of mitochondrial 12S rDNA and 16S rDNA, is supported here by the nuclear 18S rDNA dataset. The support is even stronger when the sequences of mitochondrial rDNAs and nuclear 18S rDNA are combined as a total dataset. The independent support from both nuclear 18S rDNA and mitochondrial rDNAs strongly argues for a close relationship between the Asterina and Solasteridae.

Animals↗

Gene-flow patterns in Atlantic and Mediterranean populations of the Lusitanian sea star Asterina gibbosa.

In this study, the population structure of the Lusitanian sea star Asterina gibbosa was assessed using amplified fragment length polymorphism (AFLP). One hundred and twenty-two AFLP loci were analysed in 159 individuals from eight populations from across the species' range and revealed high levels of genetic diversity, with all individuals but two harbouring a unique banding pattern. As reported for other marine invertebrates, we found high levels of genetic differentiation between the Atlantic and Mediterranean basins, suggesting that the Strait of Gibraltar represents a major barrier to dispersal for this sea star. Our assignment studies suggest that, in the Atlantic, a measurable degree of gene flow occurs between populations, which could result in the isolation-by-distance pattern of differentiation observed in this basin. In contrast, no evidence of contemporary gene flow was found in the Mediterranean, suggesting contrasting patterns of dispersal of Asterina gibbosa in the Atlantic and Mediterranean basins.

Animals↗

Two different sialidases, KDN-sialidase and regular sialidase in the starfish Asterina pectinifera.

We have found the coexistence of two different sialidases in the entrails of the starfish Asterina pectinifera: a regular sialidase (RS), which cleaves sialic acid from sialoglycoconjugates, and a KDN-sialidase (KS) which releases the sialic acid analogue KDN (2-keto-3-deoxy-D-glycero-d-galacto-nononic acid) from KDN-containing glycoconjugates that are resistant to RS. The 6700-fold purified KS and 1300-fold purified RS were prepared to study the properties of these two sialidases. KS and RS from Asterina starfish differ in several properties other than glycon specificity, including molecular mass, isoelectric point (pI) and susceptibility to competitive and non-competitive inhibitors. KS has a molecular mass of 31 kDa and a pI of 8.3 while RS has a molecular mass of 128 kDa and a pI of about 4.8. 2,3-dehydro-2-deoxy-N-acetylneuraminic acid (NeuAc2en), but not 2,3-dehydro-2-deoxy-KDN (KDN2en), is a potent competitive inhibitor of RS (Ki approximately 0.007 mM); however, both NeuAc2en and KDN2en are moderate inhibitors of KS (K1 approximately 0.04 mM). Hg2+ is a potent non-competitive inhibitor of RS but not of KS. KS and RS were examined for their ability to hydrolyse KDN- and NeuAc-containing glycoconjugates. KS hydrolyses 4-methyl-umbelliferyl-alpha-KDN (MU-KDN) 20 times faster than 4-methylumbelliferyl-alpha-NeuAc (MU-NeuAc), while RS hydrolyses MU-NeuAc 88 times faster than MU-KDN at the pH optimum of 4.0 KS effectively hydrolyses KDN-GM3 (where GM3 is NeuAc alpha 2 --> 3Gal beta 1 --> 4Glc beta 1-1' Cer, and Cer is ceramide), KDN alpha 2 --> 3lactose, KDN alpha 2 --> 6lactose, KDN alpha 2 --> 6N-acetylgalactosaminitol, KDN alpha 2 --> 6 (KDN alpha 2 --> 3)N-acetylgalactosaminitol and KDN alpha 2 --> 6(GlcNAc beta 1 --> 3) N-acetylgalactosaminitol. However, under the same conditions, these KDN-containing glycoconjugates are refractory to RS. Conversely, GM3, NeuAc alpha 2 --> 3lactose and NeuAc alpha 2 --> 6lactose are effectively hydrolysed by RS but not by KS.

Animals↗

Chemopreventive effect of protein extract of Asterina pectinifera in HT-29 human colon adenocarcinoma cells.

We investigated the effect of protein extract of Asterina pectinifera on the activity of 4 enzymes that may play a role in adenocarcinoma of the colon: quinone reductase (QR), glutathione S-transferase (GST), ornithine decarboxylase (ODC), and cyclooxygenase (COX)-2. QR and GST activity increased in HT-29 human colon adenocarcinoma cells increased that had been exposed to 4 concentrations of the protein extract (80, 160, 200, and 240 microg/mL). Additionally, 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ODC activity decreased significantly in cells exposed to the extract in concentrations of 160 microg/mL (p<0.05), 200 microg/mL (p<0.005), and 240 microg/mL (p<0.005). TPA-induced COX-2 activity also decreased in cells exposed to extract concentrations of 10, 20, 40, and 60 microg/mL. COX-2 expression was also inhibited in cells exposed to this extract. These results suggest that this protein extract of A pectinifera has chemopreventive activity in HT-29 human colon adenocarcinoma cells, and therefore, may have the potential to function as a chemopreventive agent in human colorectal cancer.

Adenocarcinoma↗

Evaluation of the attachment strength of individuals of Asterina gibbosa (Asteroidea, Echinodermata) during the perimetamorphic period.

A turbulent channel flow apparatus was used to determine the adhesion strength of the three perimetamorphic stages of the asteroid Asterina gibbosa, i.e. the brachiolaria larvae, the metamorphic individuals and the juveniles. The mean critical wall shear stresses (wall shear stress required to dislodge 50% of the attached individuals) necessary to detach larvae attached by the brachiolar arms (1.2 Pa) and juveniles attached by the tube feet (7.1 Pa) were one order of magnitude lower than the stress required to dislodge metamorphic individuals attached by the adhesive disc (41 Pa). This variability in adhesion strength reflects differences in the functioning of the adhesive organs for these different life stages of sea stars. Brachiolar arms and tube feet function as temporary adhesion organs, allowing repetitive cycles of attachment to and detachment from the substratum, whereas the adhesive disc is used only once, at the onset of metamorphosis, and is responsible for the strong attachment of the metamorphic individual, which can be described as permanent adhesion. The results confirm that the turbulent water channel apparatus is a powerful tool to investigate the adhesion mechanisms of minute organisms.

Animals↗

A novel polyhydroxyl sterol from Asterina pectinifera.

A novel polyhydroxyl sterol (1) along with one known polyhydroxyl sterol (2), and two known monoglycosides, asterosaponin P1 (3) and its desulfated monoglycoside (4), have been isolated from the whole bodies of a common Pacific starfish Asterina pectinifera. The structure of the new polyhydroxyl sterol was determined as 15beta,16beta-isopropylidenedioxy-5alpha-cholestane-3beta,4beta,6alpha,7alpha,8,26-hexaol by spectroscopic methods, including FABMS, HR-FABMS, 1D and 2D NMR techniques.

Animals↗

NGIWYamide-induced contraction of tube feet and distribution of NGIWYamide-like immunoreactivity in nerves of the starfish Asterina pectinifera.

NGIWYamide, a neuropeptide recently isolated from sea cucumbers, was tested on tube feet of the starfish Asterina pectinifera. NGIWYamide (10(-6)-10(-4) M) caused contraction of isolated tube feet. NGIWYamide-like immunoreactivity (NGIWYa-LI) was investigated with an antiserum against NGIWYamide. NGIWYa-LI was found in the radial nerve cord (RNC), the marginal nerve, and the tube feet. Both ectoneural and hyponeural parts of the RNC showed NGIWYa-LI. Immunoreactive cell bodies were found in both parts of RNC. Extensive labeling in the basal region of the ectoneural part suggests that a substantial proportion of axons in this part contains NGIWYamide or a similar substance. In tube feet, NGIWYa-LI was found in the sub-epithelial nerve plexus and in the basal nerve ring. Double labeling along with 1E11, a neuron-specific monoclonal antibody developed from A. pectinifera, indicated that the structures with NGIWYa-LI are neurons. These results suggest that NGIWYamide or an NGIWYamide-like peptide exists in starfish and functions as a neurotransmitter or a neuromodulator.

Animals↗

Adaptations to benthic development: functional morphology of the attachment complex of the brachiolaria larva in the sea star Asterina gibbosa.

The asteroid Asterina gibbosa lives all its life in close relation to the sea bottom. Indeed, this sea star possesses an entirely benthic, lecithotrophic development. The embryos adhere to the substratum due to particular properties of their jelly coat, and hatching occurs directly at the brachiolaria stage. Brachiolariae have a hypertrophied, bilobed attachment complex comprising two asymmetrical brachiolar arms and a central adhesive disc. This study aims at describing the ultrastructure of the attachment complex and possible adaptations, at the cellular level, to benthic development. Immediately after hatching, early brachiolariae attach by the arms. All along the anterior side of each arm, the epidermis encloses several cell types, such as secretory cells of two types (A and B), support cells, and sensory cells. Like their equivalents in planktotrophic larvae, type A and B secretory cells are presumably involved in a duo-glandular system in which the former are adhesive and the latter de-adhesive in function. Unlike what is observed in planktotrophic larvae, the sensory cells are unspecialized and presumably not involved in substratum testing. During the larval period, the brachiolar arms progressively increase in size and the adhesive disc becomes more prominent. At the onset of metamorphosis, brachiolariae cement themselves strongly to the substratum with the adhesive disc. The disc contains two main cell types, support cells and secretory cells, the latter being responsible for the cement release. During this metamorphosis, the brachiolar arms regress while post-metamorphic structures grow considerably, especially the tube feet, which take over the role of attachment to the substratum. The end of this period corresponds to the complete regression of the external larval structures, which also coincides with the opening of the mouth. This sequence of stages, each possessing its own adhesive strategy, is common to all asteroid species having a benthic development. In A. gibbosa, morphological adaptations to this mode of development include the hypertrophic growth of the attachment complex, its bilobed shape forming an almost completely adhesive sole, and the regression of the sensory equipment.

Animals↗

Ultrastructure of spermatogenesis in the sea star, Asterina minor.

The ultrastructural features of spermatogenesis were investigated in the hermaphroditic sea star Asterina minor. The primordial germ cells in the genital rachis contain small clusters of electron-dense material (nuage material) and a stack of annulate lamellae. They also have a flagellum and basal body complex situated close to the Golgi complex. After the development of the genital rachis into the ovotestis, spermatogenic cells increase in number and differentiation begins. Nuage material is observed in spermatogonia, but it gradually disappears in spermatocytes. The annulate lamellae do not exist beyond the early spermatogonial stage. By contrast, a flagellum and basal body complex are found throughout spermatogenesis. The Golgi-derived proacrosomal vesicles appear in the spermatocyte and coalesce to form an acrosomal vesicle in the early spermatid. The process of acrosome formation is as follows: (1) a lamella of endoplasmic reticulum (ER) continuous with the outer nuclear membrane encloses the posterior portion of the acrosomal vesicle; (2) the vesicle attaches to the cell membrane with its anterior portion; (3) periacrosomal material accumulates in the space between the acrosomal vesicle and the ER; (4) the nucleus proper changes its features to surround the acrosome; (5) amorphous, electron-dense material is deposited under the electron-dense disk; and (6) the nucleus forms a hollow opposite the electron-dense material.

Animals↗

Division of polar bodies induced by their enlargement in the starfish Asterina pectinifera.

The first polar body (FPB), which is formed at the first meiotic division during oogenesis, does not generally divide. We made a hypothesis that the amount of cytoplasm was not sufficient for the FPB to perform cell division, in spite of the same amount of genomes and centrosome as those of the secondary oocyte. To examine this hypothesis, hexylene glycol (HG) at a low concentration was applied to oocytes of the starfish Asterina pectinifera during the first meiotic division. Hence, FPBs were enlarged in their diameters, some of them divided once, and the division rate increased in proportion as their diameter extended. We further hypothesized that the difference between the second polar body (SPB) and the egg would be only the amount of cytoplasm and that if SPBs were enlarged, they would become eggs. When the secondary oocytes were treated with HG, large SPBs were obtained. Some of them, however, divided, and resultant daughter cells divided moreover, whereas eggs would not cleave unless they were fertilized. We discuss here the reason why the centrosome distributed during maturation division began to function in enlarged PBs.

Animals↗

A PCR survey of hox genes in the sea star, Asterina minor.

The sea star, Asterina minor, was surveyed for Hox genes using the method of PCR and subsequent sequence determination. Seven different Hox-type homeobox fragments and homeobox fragments of two other types, the Gbx-type and the Xlox-type, were identified. The results of comparative analysis with known homeobox sequences suggest that the sea star has only one Hox gene cluster including two genes of the anterior group, four genes of the medial group, and one gene of the posterior group. The existence of a gene of the cognate group 1 has not been known in echinoderm species. Each of the other fragments indicated a definite relationship with one of sea urchin homeoboxes. The hypothetical cluster in the sea star is consistent with the results published for another class of echinoderm, sea urchins, in the putative number of cluster. The present result provides strong evidence that a single Hox cluster is common to echinoderms and its structure in the anterior region is more similar to other deuterostomes than previously thought.

Amino Acid Sequence↗

Histofluorescence study and biochemical assay of catecholamines (dopamine and noradrenaline) during the course of arm-tip regeneration in the starfish, Asterina gibbosa (Echinodermata, Asteroidea).

In the starfish, Asterina gibbosa, the histofluorescence method gives evidence of an aminergic activity in the course of arm-tip regeneration. This activity can be detected as variations in the localization and intensity of a green fluorescence in the nerve structures of the arm, the radial nerve, and the circumoral nerve ring connected to it. Biochemical assays reveal that dopamine levels increase on the 2nd and 4th days of regeneration, when the blastema is forming and when differentiation commences. The level of noradrenaline also increases on the 2nd day after amputation of the arm.

Animals↗

Chromosomal behavior in starfish (Asterina pectinifera) zygotes under the effect of aphidicolin, an inhibitor of DNA polymerase.

When calf thymus histones were labeled fluorescently and microinjected into oocytes of the starfish, Asterina pectinifera, the labeled histones visualized chromosomes during maturation division and cleavage. In doing so, we confirmed the previously reported phenomenon that chromosomes became incompetent at the first cleavage in the aphidicolin-treated egg, although cleavage itself took place. Moreover, we found that chromosomes were aligned at the equator of the metaphase spindle of the first cleavage and that they did not separate into two groups at all, but made a lump in the middle of the spindle. Chromosomes finally entered one blastomere, although they did not participate in the following karyokinesis. DNA and microtubules were examined by cytochemistry and immunofluorescence in order to investigate the relation between chromosome movement and the microtubular cytoskeleton. The mitotic apparatus developed and grew in the aphidicolin-treated cells in the same manner as those in normal cells without normal chromatin condensation or chromosome movement during the first cleavage. However, the mitotic apparatus consisted of two asters without the spindle formed at subsequent cleavages. Electron microscopic study revealed that chromosomes did not condense normally and kinetochores were not detected during the first cleavage. These results indicate that the dynamic changes in microtubular structures during mitosis have poor relation with the chromosome behavior such as prophase chromosome condensation and anaphase chromosome movement.

Anaphase↗

Seasonal variations in the levels of polyhydroxysteroids and related glycosides in the digestive tissues of the starfish Patiria (Asterina) pectinifera.

Seasonal variations in the levels of polar steroids including polyhydroxylated steroids and related glycosides in digestive organs of the starfish Patiria (=Asterina) pectinifera have been studied. The concentration of polar steroids is related to the annual reproductive cycle of the starfish and periods of active feeding. Two peaks in concentrations of polar steroids in pyloric caeca and stomach were found, the first in winter during reorganization and the second in summer during intensive gametogenesis before spawning. Probable biological functions of polyhydroxysteroids and related glycosides are discussed. The data support the hypothesis these compounds are involved in digestion in the starfish.

Animals↗

Seasonal variations in polyhydroxysteroids and related glycosides from digestive tissues of the starfish Patiria (=Asterina) pectinifera.

Seasonal variations in the concentrations of individual polyhydroxysteroids and related low molecular weight glycosides in pyloric caeca and stomach of the starfish Patiria (=Asterina) pectinifera collected at one location near Vladivostok have been studied. HPLC analysis on the fractions containing these substances showed a fairly constant composition of steroids in digestive tissues of P. pectinifera in spite of small seasonal variations in the relative concentrations of individual compounds.

Animals↗

Pretreatment effects of jelly components on the sperm acrosome reaction and histone degradation in the starfish, Asterina pectinifera.

Acrosome reaction (AR) and histone degradation (HD) of Asterina pectinifera sperm are induced by co-operation of ARIS and a diffusible fraction (M8) of egg jelly. Once sperm are treated with ARIS or M8 separately for several minutes, they do not undergo the AR in response to the egg jelly. Preincubation of sperm with M8 at 0 degrees C is not effective to block the jelly-induced AR whereas inhibitory effects of ARIS remain at 0 degrees C. Jelly-induced HD is inhibited by pretreatment of sperm with ARIS but is not affected by the incubation with M8. The blockage of the jelly-induced reactions, both AR and HD, by ARIS- or M8-pretreatment can be bypassed by ionophores, A23187 and monensin.

Acrosome↗

Egg jelly components responsible for histone degradation and acrosome reaction in the starfish, Asterina pectinifera.

In the starfish, Asterina pectinifera, egg jelly induces the degradation of sperm histones as well as the acrosome reaction. We have isolated histone degradation-inducing components from the egg jelly. The histone degradation and the acrosome reaction are induced by a co-operative action of ARIS, which is an extremely large, sulfated glycoprotein with diffusible substance(s) in the jelly. Co-ARIS I, a steroidal saponin of the jelly, is effective to induce both reactions in the presence of ARIS.

Acrosome↗

Occurrence of paralytic shellfish poison (PSP) in the starfish Asterina pectinifera collected from the Kure Bay, Hiroshima Prefecture, Japan.

Assays were made for paralytic toxicity of marine invertebrates inhabiting at the coasts of Hiroshima Bay, where the infestation of bivalves such as cultured oysters with paralytic shellfish poison (PSP) has been occurred. The starfish Asterina pectinifera collected at the estuary of Nikoh River, Hiroshima Bay, was found to contain moderate levels of paralytic toxicity. Its highest toxicities as PSP found on July 30, 1999 were 12.5 MU/g for whole body, 11.0 MU/g for integument tissues and 3.9 MU/g for viscera, respectively. The toxicity of integument was changed from 3.6 to 11.0 MU/g in 1 year. Its paralytic toxin principles were identified as PSP toxins, composing mainly from saxitoxin (STX) group toxins such as carbamoyl-N-hydroxy neosaxitoxin (hyneoSTX), and STX, by HPLC and LC-MS, accounting for over 90 mol%. The PSP toxins contained in the starfish A. pectinifera considered to be transferred from bivalves or detritus living in the same area, which were contaminated with PSP. However, the involved pathway may be different from that of Asterias amurensis which was infested directly through food chain from its food bivalves, for its toxin pattern.

Animals↗