Mannan-Binding Lectin of the Sea Urchin Strongylocentrotus nudus

A novel lectin specific to low-branched mannans (MBL-SN) was isolated from coelomic plasma of the sea urchin Strongylocentrotus nudus by combining anion-exchange liquid chromatography on DEAE Toyopearl 650 M, affinity chromatography on mannan-Sepharose and gel filtration on the Sephacryl S-200. The molecular mass of MBL-SN was estimated by sodium dodecyl sulphate polyacrylamide gel electrophoresis under non-reducing conditions to be about 34 kDa. MBL-SN was shown to be a dimer with two identical subunits of about 17 kDa. The native MBL-SN exists as a tetramer. The physico-chemical properties of MBL-SN indicate that it belongs to C-type mannan-binding lectins. The cDNA encoding MBL-SN was cloned from the total cDNA of S. nudus coelomocytes and encodes a 17-kDa protein of 144 amino acid residues that contains a single carbohydrate-recognition domain of C-type lectins. Prediction of the MBL-SN tertiary structure using comparative modelling revealed that MBL-SN is an α/β-protein with eight β-strands and two α-helices. Comparison of the MBL-SN model with available three-dimensional structures of C-type lectins revealed that they share a common fold pattern.     

Dimorphism of Spermatozoa in the Sea Urchin Strongylocentrotus nudus

A phenomenon of dimorphism in spermatozoa has been revealed for the sea urchin Strongylocentrotus nudus. The spermatozoa are different in the configuration of the circular mitochondrion. The bulk of male gametes (73.4%) have a symmetrical mitochondrion, whereas the remainder spermatozoa (26.6%) have an asymmetrical one. No other ultrastructural differences in the structure of spermatozoa have been revealed. The two types of spermatozoa are supposed to represent different kinds of normal gametes capable of fertilization.     

Development of Serotonergic Neurons in Embryos of the Sea Urchin, Strongylocentrotus purpuratus

The development of the serotonergic component of the nervous system of larvae of S. purpuratus is traced using indirect immunofluorescence with a polyclonal antibody against the neurotransmitter serotonin. Initially one or two neuroblasts can be detected in the thickened epithelium of the animal plate of late gastrulae (56 hr). The number of immunoreactive cells increases to about eight during formation of the pluteus (85–90 hr). Immunoreactive axons appear simultaneously from all neuroblasts present in the 79 hr prism stage larva and form the apical ganglion. It is proposed that this component of the larval nervous system is derived from a small number of ectodermal cells associated with the apical tuft.     

The Time-Course of Hatching Enzyme Secretion in the Sea Urchin Strongylocentrotus purpuratus

By two independent methods, we have determined approximately the time-course of hatching enzyme secretion in the sea urchin Strongylocentrotus purpuratus . A quick-kill method indicates that a significant fraction of the enzyme is secreted between 90% and 97% of the fertilization-hatching interval. A direct assay method indicates that the remainder of the enzyme is secreted on either side of the 90–97%"window". The entire period of secretion spans from 75% to 100% or more of the fertilization-hatching interval. For embryos raised at 15°C this translates to an interval of 4.8 or more hr.     

Gastrulation in the Sea Urchin,Strongylocentrotus purpuratus,Is Disrupted by the Small Laminin Peptides YIGSR and IKVAV

Laminin is present on the apical and basolateral sides of epithelial cells of very early sea urchin blastulae. We investigated whether small laminin-peptides, known to have cell binding activities, alter the development of sea urchin embryos. The peptide YIGSR-NH₂ (850 μM) and the peptide PA22-2 (5 μM), which contains the peptide sequence IKVAV (Tashiro et al., J. Biol. Chem. 264, 16174, 1989), typically blocked archenteron formation when added to the sea water soon after fertilization. At lower doses, the YIGSR peptide allowed invagination of the archenteron but blocked archenteron extension and differentiation and evagination of the feeding arms. The effect of YIGSR and PA22-2 peptides declined when added to progressively older stages until no effect was seen when added at the mesenchyme blastula stage (24 hours after fertilization). Control peptides GRGDS, YIGSE, and SHA22, a dodeca-peptide with a scrambled IKVAV sequence, had no effect on development. The YIGSK peptide containing a conserved amino acid modification had only a small effect on gastrulation. The results suggest that YIGSR and IKVAV peptides specifically disrupt cell/extracellular matrix interactions required for normal development of the archenteron and feeding arms. Our recent finding that YTGIR is at the cell binding site of the B1 chain of S. purpuratus laminin supports this conclusion. Evidently, laminin or other laminin-like molecules are among the many extracellular matrix components needed for the invagination and extension of the archenteron during the gastrulation movements of these embryos.     

Evaluation of Sequence Variation and Selection in the Bindin Locus of the Red Sea Urchin, Strongylocentrotus franciscanus

Recent evidence suggests that gamete recognition proteins may be subjected to directed evolutionary pressure that enhances sequence variability. We evaluated whether diversity enhancing selection is operating on a marine invertebrate fertilization protein by examining the intraspecific DNA sequence variation of a 273-base pair region located at the 5′ end of the sperm bindin locus in 134 adult red sea urchins (Strongylocentrotus franciscanus). Bindin is a sperm recognition protein that mediates species-specific gamete interactions in sea urchins. The region of the bindin locus examined was found to be polymorphic with 14 alleles. Mean pairwise comparison of the 14 alleles indicates moderate sequence diversity (p-distance = 1.06). No evidence of diversity enhancing selection was found. It was not possible to reject the null hypothesis that the sequence variation observed in S. franciscanus bindin is a result of neutral evolution. Statistical evaluation of expected proportions of replacement and silent nucleotide substitutions, observed versus expected proportions of radical replacement substitutions, and conformance to the McDonald and Kreitman test of neutral evolution all indicate that random mutation followed by genetic drift created the polymorphisms observed in bindin. Observed frequencies were also highly similar to results expected for a neutrally evolving locus, suggesting that the polymorphism observed in the 5′ region of S. franciscanus bindin is a result of neutral evolution. Received: 19 June 1998 / Accepted: 2 August 2000     

Effects of Light and Covering Behavior on PAX6 Expression in the Sea Urchin Strongylocentrotus intermedius

We studied the diel expression pattern of PAX6 (a structural gene that is commonly involved in the eye development and photoreception of eye forming animals) and the effects of light and covering behavior on PAX6 expression in the sea urchin Strongylocentrotus intermedius. We confirmed that aphotic condition significantly reduced covering behavior in S. intermedius. The diel expression pattern of PAX6 was significantly different in S. intermedius under photic and aphotic conditions. The gene expression of PAX6 significantly deceased in covered S. intermedius both under natural light and in darkness. The present finding provides valuable insight into the probable link between covering and PAX6 expression of sea urchins. Further studies are required to investigate the detailed expression network of light detection involved genes in order to fully reveal the molecular mechanism of the light-induced covering behavior of sea urchins.     

Allozyme Variability of the Sea Urchin Strongylocentrotus droebachiensis in the Area of Volcanic Hydrotherms

Allozyme variability was studied in 11 polymorphic loci in three samples of the sea urchin Strongylocentrotus droebachiensis using the method of gel electrophoresis. One sample was taken from a population of normal environmental conditions (Avacha Bay, Kamchatka Peninsula), the other two were sampled in Kraternaya Bay (Yankicha Island, the Kuriles): the first sample was from a bottom site where the animals were subjected to continuous stress resulting from the products of volcanic hydrothermal vents and the second sample was from a bottom site where that stress effect was periodical. Statistically significant differences in the genotypic allele frequencies and in levels of heterozygosity of some loci were revealed between the Avacha Bay and Kraternaya Bay samples. Moreover, similar differences, but in other loci, were revealed for the Kraternaya Bay samples. The possible causes of these distinctions are discussed. A conclusion on the primary role of natural selection in the origin of the observed distinctions and on the conformity of the observed data to the hypothesis of selective significance of a part of the allozyme polymorphism is put forth.     

The Vertical Distribution of Larvae of the Sea Urchin Strongylocentrotus intermedius in Superficial Desalination Conditions

The vertical distribution of larvae of the sea urchin Strongylocentrotus intermedius in conditions of superficial desalination was studied under laboratory conditions. In the earlier stages of development, the larvae accumulated in the water column zone with a salinity below the optimal level. The long-term observations of the larval accumulations revealed the deceleration of, and abnormalities in, the development and death of the larvae. The larvae at the pre-settling stages changed their behavior: when they reached layers of a salinity unfavorable for their survival or development, the larvae did not enter them and instead moved down instead. Obviously, S. intermedius larvae are only capable of actively choosing their position within the water column with the salinity gradient at the later stages of development.     

Harvest Refugia in Marine Invertebrate Fisheries: Models and Applications to the Red Sea Urchin, Strongylocentrotus franciscanus

Benthic marine invertebrates with planktonic larvae may exhibitAllee effects in reproduction or recruitment. Hydrodynamic considerationsand experimental evidence suggest that species broadcastinggametes suffer greatly reduced fertilization efficiencies asdensities decline. There is also evidence for some species,including the red sea urchin, that post-dispersal recruitmentsuccess declines at low densities of adults, if adults provideshelter from predators or other sources of mortality. Speciesdisplaying strong Allee effects may be susceptible to catastrophicpopulation collapses with slight increases in mortality dueto harvesting or natural causes. A simple two patch logisticmodel suggests that the establishment of a harvest refuge isnecessary to prevent collapses and maintain sustainable catchesat high levels of harvesting effort. A more detailed, age structuredmodel based on the biology of the red sea urchin, Strongylocentrotusfranciscanus produces similar results. Effects of harvestingstrategies are sensitive to Allee effects caused by both fertilizationefficiency and the disruption of adult spine canopies. Theoreticalrequirements for the size and spacing of marine reserves dependsupon the dispersal abilities of the target species. Multiplereserves, spaced more closely than the average larval dispersaldistance, appear to be an effective and conservative strategyfor maintaining healthy populations and sustainable levels ofharvest.     

Sensitivity of Embryos and Larvae of the Sea Urchin Strongylocentrotus intermedius to Effects of Copper Ions

The effect of copper ions in seawater (0.02 mg/l) on the early stages of development of the sea urchin Strongylocentrotus intermedius was studied. Copper exposure from fertilization or the prism stage retarded development and growth and led to abnormalities in the morphology of the embryos and larvae. However, if development to the pluteus stage proceeded in clean seawater, an increased copper concentration did not inhibit the growth of larvae. If sea urchin embryos at fertilization and the prism stage were maintained for 1–2 days in seawater containing 0.02 mg Cu/l and then transferred to clean seawater, the adverse consequences of this exposure remained present after 48 h.     

Inheritance of Autogamy and the Killer Trait in Euplotes minuta

SYNOPSIS. Eight interfertile strains of Euplotes minuta collected from two localities in the Mediterranean Sea are shown to comprise a single breeding system of multiple mating types. One strain regularly passes through autogamy and, in crosses to non-autogamous strains, preliminary evidence for the genic control of autogamy was obtained. The cytogenetic events of conjugation and autogamy are briefly described. Three strains are "killers." Killer cells liberate a substance into the surrounding medium which has a lethal effect on cells of other strains called "sensitives." This substance, "euplotein," is not produced by sensitives. The inheritance of the killer trait is apparently under the control of a cytoplasmic factor but present attempts to identify the factor microscopically were without success. The discussion stresses the potential usefulness of this species for studies of protozoan genetics and development.     

Identification and Characterization of piRNA-Like Small RNAs in the Gonad of Sea Urchin (Strongylocentrotus nudus)

Piwi-interacting RNAs (piRNAs) and their partner PIWI proteins play an essential role in fertility, germline stem cell development, as well as the basic control and evolution of animal genomes. However, research was rare with regard to piRNA population in sea urchin, a model animal intensively used for development and genetics studies. Utilizing Solexa sequencing, we present an identification of 13,051 piRNA-like RNAs expressed in male gonad of Strongylocentrotus nudus. Out of 202 tested RNAs, 94 sequences were confirmed to express in female gonad using microarray assay, suggesting that both male and female gonads are piRNA-like RNA-enriched organs. These RNAs with "U" at the 5' end or "A" at position of 10, in size from 26 to 30 nucleotides, were predominantly 28?nt in length and tend to be clustered in small regions in genome, achieving the longest piRNA-like RNA-enriched region about 5.5?kb in scaffold78427. Alignment results showed 11 RNAs were homologous to the known piRNAs. Furthermore, BLASTn searching against sea urchin repeat element database showed these piRNA-like RNAs matched to 101 types of DNA transposons and retrotransposons, of which SPRP1, Harbinger-N2, piggyBac-N10, SINE2-1, and piggyBac-N11 were the most frequent hit elements, suggesting a transposon silencing function of these piRNA-like RNAs.     

Somatic and Oral Cortical Ultrastructure of the Plagiopylid Ciliates Lechriopyla mystax Lynch, 1930 and Plagiopyla minuta Powers, 19331

The somatic and oral cortical ultrastructure of the plagiopylid ciliates Lechriopyla mystax Lynch, 1930 and Plagiopyla minuta Powers, 1933 are described. The somatic kinetids are monokinetids with an anteriorly directed kinetodesmal fibril originating near triplets 5, 6, 7, a divergent postciliary ribbon originating at triplet 9, and an unusual transverse ribbon originating in dense material adjacent to triplets 1, 2, 3. The transverse ribbon extends beneath the right surface of the cortical ridge adjacent to the kinety from which it originated. The oral kinetids are also monokinetids from whose base rootlet fibrils extend inwards beneath the oral kineties and converge on the furcula. The striated band on these ciliates is composed of a series of short ridges orthogonal to the longitudinal axis of the band. The sides of the striated band groove are apparently supported by macrotubules. The cortical ultrastructure of the plagiopylids is discussed with reference both to the optical microscopy of the organisms and to the ultrastructure of other ciliate taxa. The plagiopylids are not clearly related to any other higher taxon and are placed incertae sedis in the Subphylum Cyrtophora Small, 1976.     

Fractal Organization of Cultivated in vitro Spiculogenous Cells and Larval Spicules of the Sea Urchin Strongylocentrotus nudus

The morphological patterns of the cultivated cells of primary mesenchyme and the spicules of the larval skeleton of the sea urchin Strongylocentrotus nudus were quantified, and the value of their fractal dimensions (D) was determined with ImageJ 1.20s software. It was shown that during cytodifferentiation, the values of D in the fractal (fractional) dimension, which reflects the complex spatial organization of the spiculogenous mesenchyme elements in two-dimensional space, increase to values close to 1.7. The invertible treatment with cytochalasin, which destroys the system of the actin filaments, suppresses the normal control of biomineralization and causes a complex form of spicules, the fractal dimension of which varies within 1.5–1.6. Thus, the determination of the fractal dimension value serves as evidence of the fractional essence of the patterns studied, quantifies the spatially complex organization of cells and their assemblies during morphogenesis, and allows us to estimate the variation in the spicule morphology after cytochalasin treatment.     

Effect of Fixed Daylengths on the Photoperiodic Regulation of Gametogenesis in the Sea Urchin Strongylocentrotus purpuratus

Summary

The gametogenic response of the sea urchin Strongylocentrotus purpuratus was compared under treatments of different photoperiod regimes of fixed and seasonal changing daylength. Quantification of gametogenic activity in en- zymatically dissagregated ovaries and in histological sections of testes have shown that the sea urchin is sensitive to both fixed and variable daylengths. After one year at fixed short day (8L:16D) or fixed neutral day (12L:12D) the gonads were ripe and in active gametogenesis, the same as were gonads of animals reared at changing photoperiod and sampled during the short-day phase of the cycle. Under fixed long day (16L:8D) the gonads lacked significant amount of gametes and had the cell constitution found in gonads of animals reared at changing photoperiod and sampled during the long-day phase of the cycle. Measurement of a critical daylength related to the autumnal equinox seems to be part of the mechanism in the photoperiodic regulation of vitellogenic oocyte growth and spermatogenesis in these animals.     

Coelomic System of the Sea Urchin Strongylocentrotus pallidus (Echinodermata,Echinoidea) and the Issue of Coelomic Metamery in Echinoderms

Doklady Biological Sciences - The representatives of the main phylogenetic clades of Bilateria are characterized by metamery. In Deuterostomia, metamery is presented in hemichordates and chordates....