Purple Sea Urchin Strongylocentrotus Purpuratus Research Articles

Self‐Sharpening Mechanism of the Sea Urchin Tooth

AbstractThe sea urchin tooth is a mosaic of calcite crystals shaped precisely into plates and fibers, cemented together by a robust calcitic polycrystalline matrix. The tooth is formed continuously at one end, while it grinds and wears at the opposite end, the sharp tip. Remarkably, these teeth enable the sea urchin to scrape and bore holes into rock, yet the teeth remain sharp rather than dull with use. Here we describe the detailed structure of the tooth of the California purple sea urchin Strongylocentrotus purpuratus, and focus on the self‐sharpening mechanism. Using high‐resolution X‐ray photoelectron emission spectromicroscopy (X‐PEEM), scanning electron microscopy (SEM), EDX analysis, nanoindentation, and X‐ray micro‐tomography, we deduce that the sea urchin tooth self‐sharpens by fracturing at discontinuities in the material. These are organic layers surrounding plates and fibers that behave as the “fault lines” in the tooth structure, as shown by nanoindentation. Shedding of tooth components at these discontinuities exposes the robust central part of the tooth, aptly termed “the stone”, which becomes the grinding tip. The precise design and position of the plates and fibers determines the profile of the tooth tip, so as the tooth wears it maintains a tip that is continually renewed and remains sharp. This strategy may be used for the top‐down or bottom‐up fabrication of lamellar materials, to be used for mechanical functions at the nano‐ and micrometer scale.

Problems of the gonad index and what can be done: analysis of the purple sea urchin Strongylocentrotus purpuratus

The gonad index, GI, is widely used as a measure of changes in reproductive state. There are, however, problems with its use because it is based on the implicit assumption of an isometric relationship between gonad size and some measure of total size. If, for example, gonad weight and total weight are used, the exponent for an allometric relationship usually is ignored and hence assumed to be 1.0. It is further assumed that this exponent is fixed for all states of the reproductive cycle and that gonads begin to develop at size = 0. Data for the purple sea urchin Strongylocentrotus purpuratus at Gregory Point, Oregon, USA, gathered over a period of 31 months showed that these assumptions cannot be supported. The relation- ship is better modeled with a function that (1) takes into account size of initial gonad production and (2) allows allometric exponents that vary with site or season. Thus, a better approach is to use a wide range of sizes to estimate size when gonads begin to develop and then, with this correction, ANCOVA to test for differences of gonad size among samples. Gonad changes at Gregory Point were estimated using fixed sizes of 5 cm diameter and 60 g total weight. Publishing means for X and Y, the standard error of the estimate, R 2 , and slope for each regression are shown to

Powerful workhorses for antimicrobial peptide expression and characterization

Discovery of antimicrobial peptides (AMP) is to a large extent based on screening of fractions of natural samples in bacterial growth inhibition assays. However, the use of bacteria is not limited to screening for antimicrobial substances. In later steps, bioengineered “bugs” can be applied to both production and characterization of AMPs. Here we describe the idea to use genetically modified Escherichia coli strains for both these purposes. This approach allowed us to investigate SpStrongylocins 1 and 2 from the purple sea urchin Strongylocentrotus purpuratus only based on sequence information from a cDNA library and without previous direct isolation or chemical synthesis of these peptides. The recombinant peptides are proved active against all bacterial strains tested. An assay based on a recombinant E. coli sensor strain expressing insect luciferase, revealed that SpStrongylocins are not interfering with membrane integrity and are therefore likely to have intracellular targets.

Purple sea urchins Strongylocentrotus purpuratus reduce grazing rates in response to risk cues from the spiny lobster Panulirus interruptus

The classical view of trophic cascades is that predators, by consuming herbivores, exert a positive indirect effect on plants. Although this form of trophic cascade has been demonstrated in a variety of terrestrial, aquatic, and marine systems, growing evidence suggests many trophic cascades are driven by anti-predator behaviors in prey. Despite abundant evidence of behavioral responses by sea urchins to predators, there has been little examination of how predation risk may influence urchin grazing rates. To determine if purple sea urchins Strongylocentrotus purpuratus graze less in the presence of predation risk, I monitored individual urchin grazing on the kelp Macrocystis pyrifera in the presence and absence of waterborne cues from damaged conspecifics and the predatory spiny lobster Panulirus interruptus. Sea urchin grazing rates were similar in the presence and absence of damaged conspecifics, but sea urchins exposed to lobster cues, regardless of lobster diet, reduced grazing rates by 44%. Given that trophic cascades involving herbivorous sea urchins exert an impor- tant influence on primary production in kelp forests, these results suggest that predation risk may play an important but under-appreciated role in the dynamics of kelp forest food webs and primary production.

Mechanism of Calcite Co-Orientation in the Sea Urchin Tooth

Sea urchin teeth are remarkable and complex calcite structures, continuously growing at the forming end and self-sharpening at the mature grinding tip. The calcite (CaCO(3)) crystals of tooth components, plates, fibers, and a high-Mg polycrystalline matrix, have highly co-oriented crystallographic axes. This ability to co-orient calcite in a mineralized structure is shared by all echinoderms. However, the physico-chemical mechanism by which calcite crystals become co-oriented in echinoderms remains enigmatic. Here, we show differences in calcite c-axis orientations in the tooth of the purple sea urchin ( Strongylocentrotus purpuratus ), using high-resolution X-ray photoelectron emission spectromicroscopy (X-PEEM) and microbeam X-ray diffraction (muXRD). All plates share one crystal orientation, propagated through pillar bridges, while fibers and polycrystalline matrix share another orientation. Furthermore, in the forming end of the tooth, we observe that CaCO(3) is present as amorphous calcium carbonate (ACC). We demonstrate that co-orientation of the nanoparticles in the polycrystalline matrix occurs via solid-state secondary nucleation, propagating out from the previously formed fibers and plates, into the amorphous precursor nanoparticles. Because amorphous precursors were observed in diverse biominerals, solid-state secondary nucleation is likely to be a general mechanism for the co-orientation of biomineral components in organisms from different phyla.

Scaling of feeding biomechanics in the horn shark Heterodontus francisci: ontogenetic constraints on durophagy

Organismal performance changes over ontogeny as the musculoskeletal systems underlying animal behavior grow in relative size and shape. As performance is a determinant of feeding ecology, ontogenetic changes in the former can influence the latter. The horn shark Heterodontus francisci consumes hard-shelled benthic invertebrates, which may be problematic for younger animals with lower performance capacities. Scaling of feeding biomechanics was investigated in H. francisci ( n=16, 19–59 cm standard length (SL)) to determine the biomechanical basis of allometric changes in feeding performance and whether this performance capacity constrains hard-prey consumption over ontogeny. Positive allometry of anterior (8–163 N) and posterior (15–382 N) theoretical bite force was attributed to positive allometry of cross-sectional area in two jaw adducting muscles and mechanical advantage at the posterior bite point (0.79–1.26). Mechanical advantage for anterior biting scaled isometrically (0.52). Fracture forces for purple sea urchins Strongylocentrotus purpuratus consumed by H. francisci ranged from 24 to 430 N. Comparison of these fracture forces to the bite force of H. francisci suggests that H. francisci is unable to consume hard prey early in its life history, but can consume the majority of S. purpuratus by the time it reaches maximum size. Despite this constraint, positive allometry of biting performance appears to facilitate an earlier entry into the durophagous niche than would an isometric ontogenetic trajectory. The posterior gape of H. francisci is significantly smaller than the urchins capable of being crushed by its posterior bite force. Thus, the high posterior bite forces of H. francisci cannot be fully utilized while consuming prey of similar toughness and size to S. purpuratus, and its potential trophic niche is primarily determined by anterior biting capacity.

Soluble Epoxide Hydrolase Homologs in Strongylocentrotus purpuratus Suggest a Gene Duplication Event and Subsequent Divergence

The mammalian soluble epoxide hydrolase (sEH) is a multidomain enzyme composed of C- and N-terminal regions that contain active sites for epoxide hydrolase (EH) and phosphatase activities, respectively. We report the cloning of two 60 kDa multidomain enzymes from the purple sea urchin Strongylocentrotus purpuratus displaying significant sequence similarity to both the N- and C-terminal domains of the mammalian sEH. While one urchin enzyme did not exhibit EH activity, the second enzyme hydrolyzed several lipid messenger molecules metabolized by the mammalian sEH, including the epoxyeicosatrienoic acids. Neither of the urchin enzymes displayed phosphatase activity. The urchin EH was inhibited by small molecule inhibitors of the mammalian sEH and is the likely ancestor of the enzyme. Sequence comparisons suggest that the urchin sEH homologs are the result of a gene fusion event between a gene encoding for an EH and a gene for an enzyme of undetermined function. This fusion event was followed by a duplication event to produce the urchin enzymes.

Population structure of purple sea urchinStrongylocentrotus purpuratusalong the Baja California peninsula

Purple sea urchin Strongylocentrotus purpuratus is fished from British Columbia, Canada to Punta Baja, Mexico. The North American population has been divided into northern and southern fishery stocks at the break of Point Conception, but little is known about its southernmost distribution along the Mexican Pacific coast of the Baja California peninsula. In this study purple sea urchin populations in six sites along the Baja California peninsula were analyzed using mitochondrial deoxyribonucleic acid restriction fragment length polymorphism (mtDNA RFLP). A homogeneous distribution of three common haplotypes among all sites was observed. A significant F ST value, however, indicated genetic structure mainly due to the haplotype array in San Miguel, Isla Todos Santos and Punta Baja sites, which were characterized by having high haplotype diversity and several unique haplotypes. Homogeneous distribution of haplotypes along the peninsula could have been influenced by the unidirectional California Current system, flowing north to south. Unique haplotypes in Punta Baja and the structure found could be the result of local oceanographic features specific to this major upwelling zone. It may be necessary to consider the Punta Baja populations individually when managing the purple sea urchin fishery in Baja California, as they show signs of being a unique stock.

Critical tissue copper residues for marine bivalve ( Mytilus galloprovincialis) and echinoderm ( Strongylocentrotus purpuratus) embryonic development: Conceptual, regulatory and environmental implications

Critical tissue copper (Cu) residues associated with adverse effects on embryo-larval development were determined for the Mediterranean mussel ( Mytilus galloprovincialis) and purple sea urchin ( Strongylocentrotus purpuratus) following laboratory exposure to Cu-spiked seawater collected from San Diego Bay, California, USA. Whole body no-observed-effect-residues (NOER) were similar, with means of 21 and 23 μg g −1 dw, for M. galloprovincialis and S. purpuratus, respectively. Mean whole body median effect residues (ER50) were 49 and 142 μg g −1 dw for M. galloprovincialis and S. purpuratus, respectively. The difference in ER50s between species was reduced to a factor of <2 when expressed as soft tissue residues. Coefficients of variation among whole body-ER50s were 3-fold lower than median waterborne effect concentrations (EC50) for both species exposed to samples varying in water quality characteristics. This suggests that tissue concentrations were a better predictor of toxicity than water concentrations. The CBRs described herein do not differentiate between the internal Cu concentrations that are metabolically available and those that are accumulated and then detoxified. They do appear, however, to be well enough related to the level of accumulation at the site of action of toxicity that they serve as useful surrogates for the copper concentration that affects embryonic development of the species tested. Results presented have potentially important implications for a variety of monitoring and assessment strategies. These include regulatory approaches for deriving saltwater ambient water quality criteria for Cu, contributions towards the development of a saltwater biotic ligand model, the conceptual approach of using CBRs, and ecological risk assessment.

The PHD domain of the sea urchin RAG2 homolog, SpRAG2L, recognizes dimethylated lysine 4 in histone H3 tails

V(D)J recombination is a somatic gene rearrangement process that assembles antigen receptor genes from individual segments during lymphocyte development. The access of the RAG1/RAG2 recombinase to these gene segments is regulated at the level of chromatin modifications, in particular histone tail modifications. Trimethylation of lysine 4 in histone H3 (H3K4me3) correlates with actively recombining gene elements, and this mark is recognized and interpreted by the plant homeodomain (PHD) of RAG2. Here we report that the PHD domain of the only known invertebrate homolog of RAG2, the SpRAG2L protein of the purple sea urchin (Strongylocentrotus purpuratus) also binds to methylated histones, but with a unique preference for H3K4me2. While the cognate substrate for the sea urchin RAG1L/RAG2L complex remains elusive, the affinity for histone tails and the nuclear localization of ectopically expressed SpRAG2L strongly support the model that this enzyme complex exerts its activity on DNA in the context of chromatin.

Toxicity of urban highway runoff with respect to storm duration

The toxicity of stormwater runoff during various time-based stages was measured in both grab and composite samples collected from three highly urbanized highway sites in Los Angeles, California between 2002 and 2005. Stormwater runoff samples were tested for toxicity using three freshwater species (the water flea Ceriodaphnia dubia, the fathead minnow Pimephales promelas, and the green algae Pseudokirchneriella subcapitatum) and two marine species (the purple sea urchin Strongylocentrotus purpuratus, and the luminescent bacteria Photobacterium phosphoreum using Microtox™). Toxicity results varied substantially throughout the storm events for both freshwater and marine species toxicity tests. In general, however, the first few samples were found to be more toxic compared with those collected during later stages of each storm event. In most cases, more than 40% of the toxicity was associated with the first 20% of discharged runoff volume. Furthermore, on average, 90% of the toxicity was observed during the first 30% of storm duration. Toxicity identification evaluation results found copper and zinc to be the primary cause of toxicity in about 90% of the samples evaluated with these procedures. Surfactants were also found to be the cause of toxicity in less than 10% of the samples.

A genomic view of TGF‐β signal transduction in an invertebrate deuterostome organism and lessons from the functional analyses of Nodal and BMP2/4 during sea urchin development

AbstractEchinoderms are marine invertebrates but are phylogenetically much more related to the chordates, to which we belong, than are other invertebrate model organisms like Drosophila or C. elegans. The recently available sequence of the sea urchin genome offers an opportunity to draw an inventory of the developmental genes represented in a non‐chordate deuterostome and to compare this complement with that present in vertebrates and other invertebrates. In the first part of this review, we draw an inventory of the genes acting in the TGF‐β signal transduction pathway and describe the sea urchin complement of ligands, receptors, Smads, extracellular modulators and transcriptional regulators present in the genome of the purple sea urchin Strongylocentrotus purpuratus. In the second part of this review, we describe the results of recent studies addressing the function of some of these TGF‐β during sea urchin embryogenesis. Finally, we discuss hypotheses regarding a possible ancestral function of the Nodal/Lefty signalling pathway in deuterostomes.

Assessing western gull predation on purple sea urchins in the rocky intertidal using optimal foraging theory

Purple sea urchins ( Strongylocentrotus purpuratus (Stimpson, 1857)) are abundant grazing invertebrates that can have a major impact on the rocky intertidal community. Predators can control the urchin population and indirectly reduce grazing activity. We determined the effects of western gull ( Larus occidentalis Audubon, 1839) predation on purple sea urchins in the rocky intertidal using the framework of optimal foraging theory and taking into account different prey-handling techniques. We recorded the foraging behavior of gulls, measured urchin availability, and estimated prey caloric content with bomb calorimetry. Western gulls selected purple sea urchins significantly more than other prey items (snails (genus Tegula Lesson, 1835), limpets (genus Collisella Dall, 1871), sea stars ( Pisaster giganteus (Stimpson, 1857) and Pisaster ochraceus (Brandt, 1835))). Larger urchins contained relatively more calories. Gulls foraged optimally when pecking by frequently selecting the most profitable size class. However, gulls chose smaller urchins than expected when air-dropping, which could have been influenced by group size and age. Gulls selected smaller purple sea urchins when foraging in larger groups likely owing to the risk of kleptoparasitism. Adults chose larger, and juveniles smaller, urchins when air-dropping, suggesting that juveniles are less experienced in foraging techniques. We estimated that gull predation could affect up to one third of the sea urchin populations locally, which could increase species diversity in the rocky intertidal community.

Ecological role of purple sea urchins.

Sea urchins are major components of marine communities. Their grazing limits algal biomass, and they are preyed upon by many predators. Purple sea urchins (Strongylocentrotus purpuratus) are among the best studied species. They live in environments that alternate between two stable states: luxuriant, species-rich kelp forests and sea urchin-dominated "barrens." The transition from one state to the other can be initiated by several factors, including the abundance of algal food, predators, storm intensities, and incidence of disease. Purple sea urchins compete with other grazers, some of which are important fishery resources (such as abalones and red sea urchins), and they are harvested for scientific research. Revelations from their genome will lead to a better understanding of how they maintain their ecological importance, and may in turn enhance their economic potential.

Intermediary metabolism in sea urchin: The first inferences from the genome sequence

The genome sequence of the purple sea urchin Strongylocentrotus purpuratus recently became available. We report the results of functional annotation and initial analysis of more than 2300 proteins predicted to be involved in metabolite transport and enzymatic conversion in sea urchin. The comparison of various reconstructed biosynthetic and catabolic pathways in sea urchin to those known in other genomes suggests the overall similarity of the sea urchin metabolism to that of the vertebrates, with relatively small but non-trivial differences from both vertebrates and protostomes. There are several examples of two parallel, non-orthologous solutions for the same molecular function in sea urchin, in contrast with the other completely sequenced metazoans that tend to contain just one version of the same function. There are also genes that appear to be close phylogenetic neighbors of plant or bacterial homologs, as opposed to homologs in other Metazoa. The evolutionary and functional significance of these variations is discussed.

A genome-wide analysis of biomineralization-related proteins in the sea urchin Strongylocentrotus purpuratus

Biomineralization, the biologically controlled formation of mineral deposits, is of widespread importance in biology, medicine, and engineering. Mineralized structures are found in most metazoan phyla and often have supportive, protective, or feeding functions. Among deuterostomes, only echinoderms and vertebrates produce extensive biomineralized structures. Although skeletons appeared independently in these two groups, ancestors of the vertebrates and echinoderms may have utilized similar components of a shared genetic “toolkit” to carry out biomineralization. The present study had two goals. First, we sought to expand our understanding of the proteins involved in biomineralization in the sea urchin, a powerful model system for analyzing the basic cellular and molecular mechanisms that underlie this process. Second, we sought to shed light on the possible evolutionary relationships between biomineralization in echinoderms and vertebrates. We used several computational methods to survey the genome of the purple sea urchin Strongylocentrotus purpuratus for gene products involved in biomineralization. Our analysis has greatly expanded the collection of biomineralization-related proteins. We have found that these proteins are often members of small families encoded by genes that are clustered in the genome. Most of the proteins are sea urchin-specific; that is, they have no apparent homologues in other invertebrate deuterostomes or vertebrates. Similarly, many of the vertebrate proteins that mediate mineral deposition do not have counterparts in the S. purpuratus genome. Our findings therefore reveal substantial differences in the primary sequences of proteins that mediate biomineral formation in echinoderms and vertebrates, possibly reflecting loose constraints on the primary structures of the proteins involved. On the other hand, certain cellular and molecular processes associated with earlier events in skeletogenesis appear similar in echinoderms and vertebrates, leaving open the possibility of deeper evolutionary relationships.

The evolution of embryonic gene expression in sea urchins

Many evolutionary modifications in development and life history derive from changes in embryonic gene expression. However, the genetic variation affecting gene expression in natural populations is not well understood, nor are the evolutionary mechanisms that operate on that variation. The early embryonic gene network of the purple sea urchin (Strongylocentrotus purpuratus) has been studied in considerable detail, providing an informative basis for analyzing the developmental and evolutionary mechanisms that alter gene expression. Comparative functional analyses have been carried out for several genes. These case studies indicate a complex relationship between sequence divergence and gene expression: in some cases, gene expression is conserved despite extensive divergence in cis-regulatory sequences, while in others the basis for a change in gene expression does not reside locally but rather in the expression or activity of transcription factors that regulate its expression. Diverse evolutionary mechanisms apparently operate on cis-regulatory regions, including negative, balancing, and stabilizing selection.

An ancient evolutionary origin of the Rag1/2 gene locus.

The diversity of antigen receptors in the adaptive immune system of jawed vertebrates is generated by a unique process of somatic gene rearrangement known as V(D)J recombination. The Rag1 and Rag2 proteins are the key mediators of this process. They are encoded by a compact gene cluster that has exclusively been identified in animal species displaying V(D)J-mediated immunity, and no homologous gene pair has been identified in other organisms. This distinctly restricted phylogenetic distribution has led to the hypothesis that one or both of the Rag genes were coopted after horizontal gene transfer and assembled into a Rag1/2 gene cluster in a common jawed vertebrate ancestor. Here, we identify and characterize a closely linked pair of genes, SpRag1L and SpRag2L, from an invertebrate, the purple sea urchin (Strongylocentrotus purpuratus) with similarity in both sequence and genomic organization to the vertebrate Rag1 and Rag2 genes. They are coexpressed during development and in adult tissues, and recombinant versions of the proteins form a stable complex with each other as well as with Rag1 and Rag2 proteins from several vertebrate species. We thus conclude that SpRag1L and SpRag2L represent homologs of vertebrate Rag1 and Rag2. In combination with the apparent absence of V(D)J recombination in echinoderms, this finding strongly suggests that linked Rag1- and Rag2-like genes were already present and functioning in a different capacity in the common ancestor of living deuterostomes, and that their specific role in the adaptive immune system was acquired much later in an early jawed vertebrate.

Thermal history-dependent expression of the hsp70 gene in purple sea urchins: Biogeographic patterns and the effect of temperature acclimation

Thermal variation associated with large spatial scales is thought to significantly affect the physiology of organisms in marine systems and consequently limit their distribution. To examine this phenomenon, this study compared the heat shock protein ( hsp70) mRNA expression of purple sea urchins ( Strongylocentrotus purpuratus) collected from different geographic locations and those acclimated to different temperatures. Tube feet were removed from S. purpuratus freshly collected from Fogarty Creek, Oregon (FC-OR) and Carpinteria, California (CRP-CA) and from urchins acclimated to 5 °C and 20 °C in the laboratory. Tissue samples were then immediately frozen or incubated at various temperatures between 5 °C and 36 °C for 30 min in vitro. Real-time PCR was used to measure the hsp70 mRNA levels of each sample. The results showed evidence for differential regulation of hsp70 between S. purpuratus in both field and laboratory studies. For example, the incubation temperature that induced maximum hsp70 mRNA expression ( T max) was higher in CRP-CA urchins (26 °C) than those from FC-OR (23 °C). Additionally, although the T max of the two acclimation groups was identical (26 °C), S. purpuratus acclimated to 5 °C failed to induce hsp70 mRNA after incubation at 32 °C, while tube feet of the 20 °C-acclimated urchins remained transcriptionally active. In general, the extent of hsp70 mRNA induction was substantial, exceeding 250-fold in the 5 °C-acclimated urchin tissues after incubation at 26 °C. These data suggest that the thermal variation encountered across S. purpuratus' range is sufficient to effect the regulation of thermally sensitive genes, like hsp70.

Cloning of a novel phospholipase C-δ isoform from pacific purple sea urchin ( Strongylocentrotus purpuratus) gametes and its expression during early embryonic development

Calcium (Ca 2+) is a ubiquitous intracellular messenger, controlling a diverse range of cellular processes, including fertilization and development of the embryo. One of the key mechanisms involved in triggering intracellular calcium release is the generation of the second messenger inositol-1,4,5-phosphate (IP 3) by the phospholipase C (PLC) class of enzymes. Although five distinct forms of PLC have been identified in mammals (β, γ, δ, ε, and ζ), only one, PLCγ, has thus far been detected in echinoderms. In the present study, we describe the isolation of a cDNA encoding a novel PLC isoform of the delta (δ) subclass, PLC-δsu, from the egg of the Pacific purple sea urchin Strongylocentrotus purpuratus. We also demonstrate the presence of this PLC within the sperm and in the early embryo. The PLC-δsu cDNA (2.44 kb) encodes a 742 amino acid polypeptide with an open reading frame of 84.6 kDa and a p I of 6.04. All of the characteristic domains found in mammalian PLCδ isoforms (PH domain, EF hands, an X–Y catalytic region, and a C2 domain) are present in PLC-δsu. A homology search revealed that PLC-δsu shares most sequence identity with bovine PLCδ2 (39%). We present evidence that PLC-δsu is expressed in unfertilized eggs, fertilized eggs, and in the early embryo. In addition to Northern and polymerase chain reaction (PCR) analyses, in situ hybridization experiments further demonstrated that the embryonic regions within which the PLC-δsu transcript can be detected during early embryonic development are associated with the highest levels of proliferative activity, suggesting a possible involvement with metabolism or cell cycle regulation.