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Sample records for abanico argopecten purpuratus

  1. Genome Sequence of Vibrio VPAP30, Isolated from an Episode of Massive Mortality of Reared Larvae of the Scallop Argopecten purpuratus

    PubMed Central

    Rojas, Rodrigo; Miranda, Claudio D.; Romero, Jaime; Asenjo, Freddy; Valderrama, Katherinne; Segovia, Cristopher

    2015-01-01

    We report here the 5.167-Mbp draft genome sequence of Vibrio VPAP30, isolated from an Argopecten purpuratus larval culture. Vibrio VPAP30 is the etiological agent of a vibriosis outbreak causing a complete collapse of a larval culture of the scallop A. purpuratus, which occurred in a commercial hatchery in Chile. PMID:26159530

  2. Genome Sequence of Vibrio VPAP30, Isolated from an Episode of Massive Mortality of Reared Larvae of the Scallop Argopecten purpuratus.

    PubMed

    Rojas, Rodrigo; Miranda, Claudio D; Romero, Jaime; Asenjo, Freddy; Valderrama, Katherinne; Segovia, Cristopher; Ugalde, Juan A; Santander, Javier

    2015-01-01

    We report here the 5.167-Mbp draft genome sequence of Vibrio VPAP30, isolated from an Argopecten purpuratus larval culture. Vibrio VPAP30 is the etiological agent of a vibriosis outbreak causing a complete collapse of a larval culture of the scallop A. purpuratus, which occurred in a commercial hatchery in Chile. PMID:26159530

  3. Genomic Characterization of Interspecific Hybrids between the Scallops Argopecten purpuratus and A. irradians irradians

    PubMed Central

    Hu, Liping; Huang, Xiaoting; Mao, Junxia; Wang, Chunde; Bao, Zhenmin

    2013-01-01

    The Peruvian scallop (Argopecten purpuratus) has been introduced to China and has successfully been hybridized with the bay scallop (A. irradians irradians). The F1 hybrids of these two scallops exhibited a large increase in production traits and some other interesting new characteristics. To understand the genetic basis of this heterosis, nuclear gene and partial mtDNA sequences, and genomic in situ hybridization (GISH) were employed to analyze the genomic organization of the hybrids. Amplification of the ribosomal DNA internal transcribed spacer (ITS) showed that the parental ITS sequences were present in all the hybrid individuals, illustrating that the hybrid offspring inherited nuclear DNA from both parents. Sequence analyses of the ITS region further confirmed that the hybrids harbored alleles from their parents; some recombinant variants were also detected, which revealed some alterations in the nuclear genetic material of the hybrids. The analysis of mitochondrial 16S rDNA showed that the hybrids possessed sequences that were identical to the 16S rDNA of the female parents, proving a matrilineal inheritance of mitochondrial genes in scallops. In addition, GISH clearly discriminated between the parental chromosomes and indicated a combination of haploid genomes of duplex parents in the hybrids. The genetic analyses in our study illustrated that the F1 hybrids inherited nuclear material from both parents and cytoplasmic genetic material maternally, and some variations occurred in the genome, which might contribute to a further understanding of crossbreeding and heterosis in scallop species. PMID:23620828

  4. Biomineralization changes with food supply confer juvenile scallops (Argopecten purpuratus) resistance to ocean acidification.

    PubMed

    Ramajo, Laura; Marbà, Núria; Prado, Luis; Peron, Sophie; Lardies, Marco A; Rodriguez-Navarro, Alejandro B; Vargas, Cristian A; Lagos, Nelson A; Duarte, Carlos M

    2016-06-01

    Future ocean acidification (OA) will affect physiological traits of marine species, with calcifying species being particularly vulnerable. As OA entails high energy demands, particularly during the rapid juvenile growth phase, food supply may play a key role in the response of marine organisms to OA. We experimentally evaluated the role of food supply in modulating physiological responses and biomineralization processes in juveniles of the Chilean scallop, Argopecten purpuratus, that were exposed to control (pH ~ 8.0) and low pH (pH ~ 7.6) conditions using three food supply treatments (high, intermediate, and low). We found that pH and food levels had additive effects on the physiological response of the juvenile scallops. Metabolic rates, shell growth, net calcification, and ingestion rates increased significantly at low pH conditions, independent of food. These physiological responses increased significantly in organisms exposed to intermediate and high levels of food supply. Hence, food supply seems to play a major role modulating organismal response by providing the energetic means to bolster the physiological response of OA stress. On the contrary, the relative expression of chitin synthase, a functional molecule for biomineralization, increased significantly in scallops exposed to low food supply and low pH, which resulted in a thicker periostracum enriched with chitin polysaccharides. Under reduced food and low pH conditions, the adaptive organismal response was to trade-off growth for the expression of biomineralization molecules and altering of the organic composition of shell periostracum, suggesting that the future performance of these calcifiers will depend on the trajectories of both OA and food supply. Thus, incorporating a suite of traits and multiple stressors in future studies of the adaptive organismal response may provide key insights on OA impacts on marine calcifiers. PMID:26644007

  5. The role of Argopecten purpuratus shells structuring the soft bottom community in shallow waters of southern Peru

    NASA Astrophysics Data System (ADS)

    Lomovasky, Betina J.; Gamero, Patricia A.; Romero, Leonardo; Firstater, Fausto N.; Gamarra Salazar, Alex; Hidalgo, Fernando; Tarazona, Juan; Iribarne, Oscar O.

    2015-12-01

    Accumulation of Argopecten purpuratus shells often occurs after El Niño events in shallow waters of Independencia Bay (14°17‧S-76°10‧W; Pisco, Peru). Here we experimentally investigate the effects of their shell accumulation on macrobenthos assemblages in soft bottom, shallow areas of the bay. A field experiment (from May 2006 to May 2007), including four treatments with different coverage levels of empty shells of A. purpuratus, were randomly arranged in: (1) areas devoid of shells ("Empty" treatment: experimental control), (2) 50% of the plot area covered with shells haphazardly distributed over the bottom ("medium" treatment), (3) 100% of the plot area covered with shells, forming a 10 cm valve layer ("full" treatment) and (4) "natural control". We found a total of 124 taxa throughout the experiment. Polychaetes, crustaceans and mollusks were the most abundant groups in "natural controls", dominated by the gastropod Nassarius gayi and the polychaetes Prionospio peruana, Platynereis bicanaliculata and Mediomastus branchiferus. The abundance of individuals (N) and the species richness (S) were higher in the "medium" treatment, but only in one month under positive sea bottom thermal anomalies. Similarity analysis (Bray-Curtis) showed that "natural control", "empty" and "full" treatments were more similar among them than the "medium" treatment. Multidimensional analysis showed no clear species association among treatments and a higher grouping among the samplings of Jun-06, Aug-06 and Nov-06. Our results also showed that the commercial crab Romaleon polyodon and the polyplacophora Tonicia elegans were positively affected by shell accumulations ("medium" treatment), while the limpet Fissurella crassa was negatively affected. Our study shows that directly by changing habitat structure or indirectly by changing sediment characteristics, the addition of scallop shells to the soft bottom can modify the macrobenthic assemblage; however, the seasonal oceanographic

  6. Tissue-specific Cd and Pb accumulation in Peruvian scallop (Argopecten purpuratus) transplanted to a suspended and bottom culture at Sechura Bay, Peru.

    PubMed

    Loaiza, Iván; Hurtado, Daniela; Miglio, Maria; Orrego, Henry; Mendo, Jaime

    2015-02-28

    In order to understand the effect of different culture systems on Cd and Pb accumulation, suspended long-line and bottom cultures of Argopecten purpuratus were conducted during January until April 2010 (120 days). The Cd tissue levels were the highest at the middle of the experiment (30-d till 70-d) for suspended-cultured individuals, while bottom-cultured individuals showed an increasing trend. Gonad Pb levels were also higher during the same period for all cultures, while adductor muscle exhibited no considerable variations. Cd and Pb tissue concentrations were mainly greater in deeper cultures. There were no significant differences in Cd and Pb accumulation between individual sizes. The Cd and Pb levels in edible tissue (gonad+adductor muscle) did not exceed the EU and FDA maximum levels. Based on the target hazard quotient (THQ) and the provisional tolerance weekly intake (PTWI), no risk (THQ<1 and %PTWI<30) was found for human consumption. PMID:25444617

  7. Molecular characterization of two ferritins of the scallop Argopecten purpuratus and gene expressions in association with early development, immune response and growth rate.

    PubMed

    Coba de la Peña, Teodoro; Cárcamo, Claudia B; Díaz, María I; Brokordt, Katherina B; Winkler, Federico M

    2016-08-01

    Ferritin is involved in several iron homoeostasis processes in molluscs. We characterized two ferritin homologues and their expression patterns in association with early development, growth rate and immune response in the scallop Argopecten purpuratus, a species of economic importance for Chile and Peru. Two ferritin subunits (Apfer1 and Apfer2) were cloned. Apfer1 cDNA is a 792bp clone containing a 516bp open reading frame (ORF) that corresponds to a novel ferritin subunit in A. purpuratus. Apfer2 cDNA is a 681bp clone containing a 522bp ORF that corresponds to a previously sequenced EST. A putative iron responsive element (IRE) was identified in the 5'-untranslated region of both genes. The deduced protein sequences of both cDNAs possessed the motifs and domains characteristic of functional ferritin subunits. Both genes showed differential expression patterns at tissue-specific and early development stage levels. Apfer1 expression level increased 40-fold along larval developmental stages, decreasing markedly after larval settlement. Apfer1 expression in mantle tissue was 2.8-fold higher in fast-growing than in slow-growing scallops. Apfer1 increased 8-fold in haemocytes 24h post-challenge with the bacterium Vibrio splendidus. Apfer2 expression did not differ between fast- and slow-growing scallops or in response to bacterial challenge. These results suggest that Apfer1 and Apfer2 may be involved in iron storage, larval development and shell formation. Apfer1 expression may additionally be involved in immune response against bacterial infections and also in growth; and thus would be a potential marker for immune capacity and for fast growth in A. purpuratus. PMID:27040527

  8. Sclerochronological records and daily microgrowth of the Peruvian scallop (Argopecten purpuratus, Lamarck, 1819) related to environmental conditions in Paracas Bay, Pisco, Peru

    NASA Astrophysics Data System (ADS)

    Aguirre Velarde, Arturo; Flye-Sainte-Marie, Jonathan; Mendo, Jaime; Jean, Fred

    2015-05-01

    We investigated the rhythm of micro-striae formation in the shell of Argopecten purpuratus and environmental influence on micro-growth increments by monitoring growth over a 98-day period between April and July 2007 under bottom and suspended culture (2 m above the bottom) rearing conditions. The transfer of individuals to the study site induced the formation of a notable growth mark that allowed us to count the number of micro-striae formed between transfer and sampling dates. Micro-striae counts showed a deposition rate of one stria per day independent of rearing condition. This result allowed us to analyse the relationships between growth increments and environmental conditions. We therefore examined the deviations between observed growth rates and growth rates predicted from a Von Bertalanffy growth function. Cross-correlation analysis revealed significant correlations, without time-lag, between these deviations and both particulate organic carbon and nitrogen concentrations in the bottom treatment. Additionally, we observed negative correlations with temperature and current speed at this depth with time-lags of 1 and 10 days respectively. In the suspended treatment, we observed a significant negative correlation with temperature, only with a 12-day lag-time. Our results show that growth response to environmental variability is not always instantaneous. This delay can be explained by the time delay over which metabolic processes need to be performed (e.g. digestion, use/movements of reserves, growth, reproduction). Further modeling studies could help to better understand these processes.

  9. Effect of diet and temperature upon muscle metabolic capacities and biochemical composition of gonad and muscle in Argopecten purpuratus Lamarck 1819.

    PubMed

    Martínez; Brokordt; Aguilera; Soto; Guderley

    2000-04-26

    Recently spawned Argopecten purpuratus broodstock were conditioned at two temperatures and fed three different diets (microalgae, microalgae mixed with lipids and microalgae mixed with carbohydrates) to examine changes in the biochemical composition of gonad and muscle as well as muscle metabolic capacities. During one experiment, scallops were fed at 3% of their dry mass per day whereas during a second experiment, they were fed at 6% of their dry mass per day. During both experiments, total gonadal levels of lipids and protein increased markedly during conditioning with the two mixed diets at 16 degrees C. These increases were less pronounced at 20 degrees C. Carbohydrate gonadal levels only increased during the second experiment at both temperatures and with the three diets. Of the major biochemical components of the adductor muscle, carbohydrate levels changed most during conditioning. Whereas muscle protein levels increased slightly with gonadal maturation, carbohydrate levels dropped considerably. Despite the marked drop in the levels of glycolytic substrates, only the activities of octopine dehydrogenase in the adductor muscle of the scallops conditioned at 16 degrees C consistently decreased. Muscle levels of glycogen phosphorylase were higher in mature than in recently spawned (control) scallops, suggesting a role in the transfer of glucose equivalents from the adductor muscle to other tissues. PMID:10727686

  10. Characterization and pathogenicity of Vibrio splendidus strains associated with massive mortalities of commercial hatchery-reared larvae of scallop Argopecten purpuratus (Lamarck, 1819).

    PubMed

    Rojas, Rodrigo; Miranda, Claudio D; Opazo, Rafael; Romero, Jaime

    2015-01-01

    Three strains (VPAP16, VPAP18 and VPAP23 strains) were isolated as the most predominant organisms from 3 different episodes of massive mortalities of larval cultures of the Chilean scallop Argopecten purpuratus occurred in different commercial hatcheries located in northern Chile. The main aims of this study were to identify the pathogenic strains and investigate their pathogenic activity. Based on selected phenotypic features and sequence identity of the 16S rRNA gene and the housekeeping gene, RNA polymerase α-chain rpoA, all pathogenic strains were identified as Vibrio splendidus. Healthy 10-day-old scallop larvae cultures exhibited mortality percentages of 69.61±3.35%, 79.78±6.11% and 61.73±3.71% after 48 h when were inoculated with 1×10(6) CFU (colony forming units)mL(-1) of VPAP16, VPAP18 and VPAP23 strains, respectively, and evidenced that concentrations ⩾10(4) CFU mL(-1) would probably be detrimental for the larval culture. The main clinical signs observed in challenged larvae for 24h were bacterial swarms on the margins of the larvae, extension and disruption of the velum, detachment of velum cilia cells and digestive tissue necrosis. Otherwise, challenge assays using pathogenic strains stained with 5-([4,6-dichlorotriazin-2-yl]amino)fluorescein hydrochloride (5-DTAF)evidenced that after 1h stained bacteria were detected in high density in the digestive gland and the margin of the shell. When larval cultures were inoculated with cell-free extracellular products (ECP) of V. splendidus strains, exhibited larval mortalities higher than 70% (VPAP16), 80% (VPAP18) and 50% (VPAP23) after 24 h, even when ECP were treated with proteinase K or heat, indicating that extracellular pathogenic activity is mainly mediated by non-proteic thermostable compounds. In this study all Koch's postulates were fulfilled and it was demonstrated for the first time the pathogenic activity of V. splendidus strains on reared-larvae of scallop A. purpuratus and prompt the

  11. Molecular characterization of an inhibitor of NF-κB in the scallop Argopecten purpuratus: First insights into its role on antimicrobial peptide regulation in a mollusk.

    PubMed

    Oyanedel, D; Gonzalez, R; Flores-Herrera, P; Brokordt, K; Rosa, R D; Mercado, L; Schmitt, P

    2016-05-01

    Inhibitors of nuclear factor kappa B (IκBs) are major control components of the Rel/NF-κB signaling pathway, a key regulator in the modulation of the expression of immune-related genes in vertebrates and invertebrates. The activation of the Rel/NF-κB signaling pathway depends largely in the degradation of IκB proteins and thus, IκBs are a main target for the identification of genes whose expression is controlled by Rel/NF-κB pathway. In order to identify such regulation in bivalve mollusks, the cDNA sequence encoding an IκB protein was characterized in the scallop Argopecten purpuratus, ApIκB. The cDNA sequence of ApIκB is comprised of 1480 nucleotides with a 1086 bp open reading frame encoding for 362 amino acids. Bioinformatics analysis showed that ApIκB displays the conserved features of IκB proteins. The deduced amino acid sequence consists of a 39.7 kDa protein, which has an N-terminal degradation motif, six ankyrin repeats and a C-terminal phosphorylation site motif. Phylogenetic analysis revealed a high degree of identity between ApIκB and other IκBs from mollusks, but also to arthropod cactus proteins and vertebrate IκBs. Tissue expression analysis indicated that ApIκB is expressed in all examined tissues and it is upregulated in circulating hemocytes from scallops challenged with the pathogenic Gram-negative bacterium Vibrio splendidus. After inhibiting ApIκB gene expression using the RNA interference technology, the gene expression of the antimicrobial peptide big defensin was upregulated in hemocytes from non-challenged scallops. Results suggest that ApIκB may control the expression of antimicrobial effectors such as big defensin via a putative Rel/NF-κB signaling pathway. This first evidence will help to deepen the knowledge of the Rel/NF-κB conserved pathway in scallops. PMID:26993612

  12. Fanning the Optimal Breeze with an Abanico

    NASA Astrophysics Data System (ADS)

    Goon, Grace; Marthelot, Joel; Reis, Pedro; MIT EGS Lab Team

    Flexible hand-held fans, or abanicos, are universally employed as cooling devices that are both portable and sustainable. Their to and fro axial motion about one's hand generates an airflow that increases the evaporation rate near the skin and refreshes. We study this problem in the context of fluid-structure interaction, through precision model experiments. We first characterize the elastic properties of a semi-circular thin plates with various thickness and evaluate their aerodynamic performance in a custom built apparatus. The air velocity profile that results from the flapping motion of the fan is characterized for different driving conditions. A systematic variation of the geometric and elastic parameters, along with an exploration of the parameter space of the periodic driving motion (amplitude and frequency), allows us to establish optimal design and operational conditions for maximal output of the generated airflow, while minimizing the input power.

  13. Fanning the Optimal Breeze with an Abanico

    NASA Astrophysics Data System (ADS)

    Goon, Grace; Marthelot, Joel; Reis, Pedro

    2015-11-01

    Flexible hand-held fans, or abanicos, are universally employed as cooling devices that are both portable and sustainable. Their to and fro axial motion about one's hand generates an airflow that increases the evaporation rate near the skin and refreshes. We study this problem in the context of fluid-structure interaction, through precision model experiments. We first characterize the elastic properties of a semi-circular thin plates with various thickness and evaluate their aerodynamic performance in a custom built apparatus. The air velocity profile that results from the flapping motion of the fan is characterized for different driving conditions. A systematic variation of the geometric and elastic parameters, along with an exploration of the parameter space of the periodic driving motion (amplitude and frequency), allows us to establish optimal design and operational conditions for maximal output of the generated airflow, while minimizing the input power.

  14. JUVENILE BAY SCALLOP (ARGOPECTEN IRRADIANS) HABITAT PREFERENCES

    EPA Science Inventory

    Habitat quality and quantity are known to be important for maintaining populations of bay scallops (Argopecten irradians), but data linking habitat attributes to bay scallop populations are lacking. This information is essential to understand the role of habitat alteration in th...

  15. Fanning the Optimal Breeze with an Abanico

    NASA Astrophysics Data System (ADS)

    Goon, Grace; Marthelot, Joel; Reis, Pedro

    2015-03-01

    Flexible hand-held fans, or abanicos, are universally employed as cooling devices that are both portable and sustainable. Their to and fro axial motion about one's hand generates an airflow that increases the evaporation rate near the skin and refreshes. We study this problem in the context of fluid-structure interaction, through precision model experiments. We first characterize the elastic properties of a number of commercially available fans and evaluate their aerodynamic performance in a custom built apparatus. The air velocity profile that results from the flapping motion of the fan is characterized for different driving conditions. We then fabricate our own analogue model fans that comprise a thin elastic plate, shaped as a circular section, with an underlying substructure of radial slats. A systematic variation of the geometric and elastic parameters, along with an exploration of the parameter space of the periodic driving motion (amplitude and frequency), allows us to establish optimal design and operational conditions for maximal output of the generated airflow, while minimizing the input power.

  16. A HABITAT SUITABILITY INDEX FOR THE BAY SCALLOP ARGOPECTEN IRRADIANS

    EPA Science Inventory

    Platform Presentation at a Scientific Meeting. A survey of Lagoon Pond, Martha's Vineyard, MA, USA was conducted in September 2005 to determine the combination of habitat factors most highly correlated with bay scallop (Argopecten irradians) abundance. A stratified random samplin...

  17. HABITAT ASSESSMENT MODELS FOR BAY SCALLOP, ARGOPECTEN IRRADIANS

    EPA Science Inventory

    Bay scallops (Argopecten irradians) inhabit shallow subtidal habitats along the Atlantic coast of the United States and require settlement substrates, such as submerged aquatic vegetation (SAV), for their early juvenile stages. The short lifespan of bay scallops (1-2 yr) coupled...

  18. A HABITAT SUITABILITY INDEX FOR THE BAY SCALLOP ARGOPECTEN IRRADIANS

    EPA Science Inventory

    Poster presentation at a scientific meeting. A survey of Lagoon Pond, Martha's Vineyard, MA, USA was conducted in September 2005 to determine the combination of habitat factors most highly correlated with the bay scallop Argopecten irradians) abundance. A stratified random samp...

  19. The complete mitochondrial genome of Saxidomus purpuratus (Veneroida: Veneridae).

    PubMed

    Bao, Xiangbo; He, Chongbo; Gao, Xianggang; Li, Yunfeng; Gao, Lei; Jiang, Bing; Liu, Weidong

    2016-09-01

    In this study, the complete mitochondrial genome of Saxidomus purpuratus is determined, which is the first complete mitochondrial genome in the genus Saxidomus. The genome was of 19 637 bp in length, including 2 rRNAs, 22 tRNAs and 12 protein-coding genes with the order of ND3 and ND5 reversed. Maximum likelihood tree based on nucleotide sequences of 12 mitochondrial PCGs was constructed, in which S. purpuratus was clustered with 3 Meretrix species. The results are expected to provide useful data for species identification and further studies of the genus Saxidomus. PMID:26329595

  20. The genome of the sea urchin Strongylocentrotus purpuratus.

    PubMed

    Sodergren, Erica; Weinstock, George M; Davidson, Eric H; Cameron, R Andrew; Gibbs, Richard A; Angerer, Robert C; Angerer, Lynne M; Arnone, Maria Ina; Burgess, David R; Burke, Robert D; Coffman, James A; Dean, Michael; Elphick, Maurice R; Ettensohn, Charles A; Foltz, Kathy R; Hamdoun, Amro; Hynes, Richard O; Klein, William H; Marzluff, William; McClay, David R; Morris, Robert L; Mushegian, Arcady; Rast, Jonathan P; Smith, L Courtney; Thorndyke, Michael C; Vacquier, Victor D; Wessel, Gary M; Wray, Greg; Zhang, Lan; Elsik, Christine G; Ermolaeva, Olga; Hlavina, Wratko; Hofmann, Gretchen; Kitts, Paul; Landrum, Melissa J; Mackey, Aaron J; Maglott, Donna; Panopoulou, Georgia; Poustka, Albert J; Pruitt, Kim; Sapojnikov, Victor; Song, Xingzhi; Souvorov, Alexandre; Solovyev, Victor; Wei, Zheng; Whittaker, Charles A; Worley, Kim; Durbin, K James; Shen, Yufeng; Fedrigo, Olivier; Garfield, David; Haygood, Ralph; Primus, Alexander; Satija, Rahul; Severson, Tonya; Gonzalez-Garay, Manuel L; Jackson, Andrew R; Milosavljevic, Aleksandar; Tong, Mark; Killian, Christopher E; Livingston, Brian T; Wilt, Fred H; Adams, Nikki; Bellé, Robert; Carbonneau, Seth; Cheung, Rocky; Cormier, Patrick; Cosson, Bertrand; Croce, Jenifer; Fernandez-Guerra, Antonio; Genevière, Anne-Marie; Goel, Manisha; Kelkar, Hemant; Morales, Julia; Mulner-Lorillon, Odile; Robertson, Anthony J; Goldstone, Jared V; Cole, Bryan; Epel, David; Gold, Bert; Hahn, Mark E; Howard-Ashby, Meredith; Scally, Mark; Stegeman, John J; Allgood, Erin L; Cool, Jonah; Judkins, Kyle M; McCafferty, Shawn S; Musante, Ashlan M; Obar, Robert A; Rawson, Amanda P; Rossetti, Blair J; Gibbons, Ian R; Hoffman, Matthew P; Leone, Andrew; Istrail, Sorin; Materna, Stefan C; Samanta, Manoj P; Stolc, Viktor; Tongprasit, Waraporn; Tu, Qiang; Bergeron, Karl-Frederik; Brandhorst, Bruce P; Whittle, James; Berney, Kevin; Bottjer, David J; Calestani, Cristina; Peterson, Kevin; Chow, Elly; Yuan, Qiu Autumn; Elhaik, Eran; Graur, Dan; Reese, Justin T; Bosdet, Ian; Heesun, Shin; Marra, Marco A; Schein, Jacqueline; Anderson, Michele K; Brockton, Virginia; Buckley, Katherine M; Cohen, Avis H; Fugmann, Sebastian D; Hibino, Taku; Loza-Coll, Mariano; Majeske, Audrey J; Messier, Cynthia; Nair, Sham V; Pancer, Zeev; Terwilliger, David P; Agca, Cavit; Arboleda, Enrique; Chen, Nansheng; Churcher, Allison M; Hallböök, F; Humphrey, Glen W; Idris, Mohammed M; Kiyama, Takae; Liang, Shuguang; Mellott, Dan; Mu, Xiuqian; Murray, Greg; Olinski, Robert P; Raible, Florian; Rowe, Matthew; Taylor, John S; Tessmar-Raible, Kristin; Wang, D; Wilson, Karen H; Yaguchi, Shunsuke; Gaasterland, Terry; Galindo, Blanca E; Gunaratne, Herath J; Juliano, Celina; Kinukawa, Masashi; Moy, Gary W; Neill, Anna T; Nomura, Mamoru; Raisch, Michael; Reade, Anna; Roux, Michelle M; Song, Jia L; Su, Yi-Hsien; Townley, Ian K; Voronina, Ekaterina; Wong, Julian L; Amore, Gabriele; Branno, Margherita; Brown, Euan R; Cavalieri, Vincenzo; Duboc, Véronique; Duloquin, Louise; Flytzanis, Constantin; Gache, Christian; Lapraz, François; Lepage, Thierry; Locascio, Annamaria; Martinez, Pedro; Matassi, Giorgio; Matranga, Valeria; Range, Ryan; Rizzo, Francesca; Röttinger, Eric; Beane, Wendy; Bradham, Cynthia; Byrum, Christine; Glenn, Tom; Hussain, Sofia; Manning, Gerard; Miranda, Esther; Thomason, Rebecca; Walton, Katherine; Wikramanayke, Athula; Wu, Shu-Yu; Xu, Ronghui; Brown, C Titus; Chen, Lili; Gray, Rachel F; Lee, Pei Yun; Nam, Jongmin; Oliveri, Paola; Smith, Joel; Muzny, Donna; Bell, Stephanie; Chacko, Joseph; Cree, Andrew; Curry, Stacey; Davis, Clay; Dinh, Huyen; Dugan-Rocha, Shannon; Fowler, Jerry; Gill, Rachel; Hamilton, Cerrissa; Hernandez, Judith; Hines, Sandra; Hume, Jennifer; Jackson, Laronda; Jolivet, Angela; Kovar, Christie; Lee, Sandra; Lewis, Lora; Miner, George; Morgan, Margaret; Nazareth, Lynne V; Okwuonu, Geoffrey; Parker, David; Pu, Ling-Ling; Thorn, Rachel; Wright, Rita

    2006-11-10

    We report the sequence and analysis of the 814-megabase genome of the sea urchin Strongylocentrotus purpuratus, a model for developmental and systems biology. The sequencing strategy combined whole-genome shotgun and bacterial artificial chromosome (BAC) sequences. This use of BAC clones, aided by a pooling strategy, overcame difficulties associated with high heterozygosity of the genome. The genome encodes about 23,300 genes, including many previously thought to be vertebrate innovations or known only outside the deuterostomes. This echinoderm genome provides an evolutionary outgroup for the chordates and yields insights into the evolution of deuterostomes. PMID:17095691

  1. The Genome of the Sea Urchin Strongylocentrotus purpuratus

    PubMed Central

    2011-01-01

    We report the sequence and analysis of the 814-megabase genome of the sea urchin Strongylocentrotus purpuratus, a model for developmental and systems biology. The sequencing strategy combined whole-genome shotgun and bacterial artificial chromosome (BAC) sequences. This use of BAC clones, aided by a pooling strategy, overcame difficulties associated with high heterozygosity of the genome. The genome encodes about 23,300 genes, including many previously thought to be vertebrate innovations or known only outside the deuterostomes. This echinoderm genome provides an evolutionary outgroup for the chordates and yields insights into the evolution of deuterostomes. PMID:17095691

  2. Spatial vision in the purple sea urchin Strongylocentrotus purpuratus (Echinoidea).

    PubMed

    Yerramilli, D; Johnsen, S

    2010-01-15

    Recent evidence that echinoids of the genus Echinometra have moderate visual acuity that appears to be mediated by their spines screening off-axis light suggests that the urchin Strongylocentrotus purpuratus, with its higher spine density, may have even more acute spatial vision. We analyzed the movements of 39 specimens of S. purpuratus after they were placed in the center of a featureless tank containing a round, black target that had an angular diameter of 6.5 deg. or 10 deg. (solid angles of 0.01 sr and 0.024 sr, respectively). An average orientation vector for each urchin was determined by testing the animal four times, with the target placed successively at bearings of 0 deg., 90 deg., 180 deg. and 270 deg. (relative to magnetic east). The urchins showed no significant unimodal or axial orientation relative to any non-target feature of the environment or relative to the changing position of the 6.5 deg. target. However, the urchins were strongly axially oriented relative to the changing position of the 10 deg. target (mean axis from -1 to 179 deg.; 95% confidence interval +/- 12 deg.; P<0.001, Moore's non-parametric Hotelling's test), with 10 of the 20 urchins tested against that target choosing an average bearing within 10 deg. of either the target center or its opposite direction (two would be expected by chance). In addition, the average length of the 20 target-normalized bearings for the 10 deg. target (each the vector sum of the bearings for the four trials) were far higher than would be expected by chance (P<10(-10); Monte Carlo simulation), showing that each urchin, whether it moved towards or away from the target, did so with high consistency. These results strongly suggest that S. purpuratus detected the 10 deg. target, responding either by approaching it or fleeing it. Given that the urchins did not appear to respond to the 6.5 deg. target, it is likely that the 10 deg. target was close to the minimum detectable size for this species. Interestingly

  3. Structure and mechanical properties of Saxidomus purpuratus biological shells.

    PubMed

    Yang, W; Zhang, G P; Zhu, X F; Li, X W; Meyers, M A

    2011-10-01

    The strength and fracture behavior of Saxidomus purpuratus shells were investigated and correlated with the structure. The shells show a crossed lamellar structure in the inner and middle layers and a fibrous/blocky and porous structure composed of nanoscaled particulates (~100 nm diameter) in the outer layer. It was found that the flexure strength and fracture mode are a function of lamellar organization and orientation. The crossed lamellar structure of this shell is composed of domains of parallel lamellae with approximate thickness of 200-600 nm. These domains have approximate lateral dimensions of 10-70 μm with a minimum of two orientations of lamellae in the inner and middle layers. Neighboring domains are oriented at specific angles and thus the structure forms a crossed lamellar pattern. The microhardness across the thickness was lower in the outer layer because of the porosity and the absence of lamellae. The tensile (from flexure tests) and compressive strengths were analyzed by means of Weibull statistics. The mean tensile (flexure) strength at probability of 50%, 80-105 MPa, is on the same order as the compressive strength (~50-150 MPa) and the Weibull moduli vary from 3.0 to 7.6. These values are significantly lower than abalone nacre, in spite of having the same aragonite structure. The lower strength can be attributed to a smaller fraction of the organic interlayer. The fracture path in the specimens is dominated by the orientation of the domains and proceeds preferentially along lamella boundaries. It also correlates with the color changes in the cross section of the shell. The cracks tend to undergo a considerable change in orientation when the color changes abruptly. The distributions of strengths, cracking paths, and fracture surfaces indicate that the mechanical properties of the shell are anisotropic with a hierarchical nature. PMID:21783161

  4. Presence of two mitochondrial genomes in the mytilid Perumytilus purpuratus: Phylogenetic evidence for doubly uniparental inheritance

    PubMed Central

    Vargas, Jaime; Pérez, Montse; Toro, Jorge; Astorga, Marcela P.

    2015-01-01

    This study presents evidence, using sequences of ribosomal 16S and COI mtDNA, for the presence of two mitochondrial genomes in Perumytilus purpuratus. This may be considered evidence of doubly uniparental mtDNA inheritance. The presence of the two types of mitochondrial genomes differentiates females from males. The F genome was found in the somatic and gonadal tissues of females and in the somatic tissues of males; the M genome was found in the gonads and mantle of males only. For the mitochondrial 16S region, ten haplotypes were found for the F genome (nucleotide diversity 0.004), and 7 haplotypes for the M genome (nucleotide diversity 0.001), with a distance Dxy of 0.125 and divergence Kxy of 60.33%. For the COI gene 17 haplotypes were found for the F genome (nucleotide diversity 0.009), and 10 haplotypes for the M genome (nucleotide diversity 0.010), with a genetic distance Dxy of 0.184 and divergence Kxy of 99.97%. Our results report the presence of two well-differentiated, sex-specific types of mitochondrial genome (one present in the male gonad, the other in the female gonad), implying the presence of DUI in P. purpuratus. These results indicate that care must be taken in phylogenetic comparisons using mtDNA sequences of P. purpuratus without considering the sex of the individuals. PMID:26273220

  5. Presence of two mitochondrial genomes in the mytilid Perumytilus purpuratus: Phylogenetic evidence for doubly uniparental inheritance.

    PubMed

    Vargas, Jaime; Pérez, Montse; Toro, Jorge; Astorga, Marcela P

    2015-05-01

    This study presents evidence, using sequences of ribosomal 16S and COI mtDNA, for the presence of two mitochondrial genomes in Perumytilus purpuratus. This may be considered evidence of doubly uniparental mtDNA inheritance. The presence of the two types of mitochondrial genomes differentiates females from males. The F genome was found in the somatic and gonadal tissues of females and in the somatic tissues of males; the M genome was found in the gonads and mantle of males only. For the mitochondrial 16S region, ten haplotypes were found for the F genome (nucleotide diversity 0.004), and 7 haplotypes for the M genome (nucleotide diversity 0.001), with a distance Dxy of 0.125 and divergence Kxy of 60.33%. For the COI gene 17 haplotypes were found for the F genome (nucleotide diversity 0.009), and 10 haplotypes for the M genome (nucleotide diversity 0.010), with a genetic distance Dxy of 0.184 and divergence Kxy of 99.97%. Our results report the presence of two well-differentiated, sex-specific types of mitochondrial genome (one present in the male gonad, the other in the female gonad), implying the presence of DUI in P. purpuratus. These results indicate that care must be taken in phylogenetic comparisons using mtDNA sequences of P. purpuratus without considering the sex of the individuals. PMID:26273220

  6. Effect of the Algaecide Palmitoleic Acid on the Immune Function of the Bay Scallop Argopecten irradians.

    PubMed

    Chi, Cheng; Giri, Sib Sankar; Jun, Jin Woo; Kim, Hyoun Joong; Kim, Sang Guen; Yun, Saekil; Park, Se Chang

    2016-01-01

    Palmitoleic acid (PA), an algicidal compound, is used against the toxin producing dinofagelate Alexandrium tamarense, however, its impact on the edible bay scallop (Argopecten irradians) is still unclear. Therefore, we investigated the impacts of effective algicidal concentrations (20, 40, and 80 mg/L) of PA on immune responses in A. irradians. Various immune parameters including acid phosphatase (ACP) activity, superoxide dismutase (SOD), lysozyme, phagocytic activity, total protein, malondialdehyde (MDA) level, and reactive oxygen species (ROS) production and the expression of immune-related genes (PrxV, CLT-6, MT, and BD) were measured at 3, 6, 12, 24, and 48 h post-exposure (hpe) to PA. Lysozyme activity was lower in scallops at 12-48 hpe to 80 mg/L. SOD, ACP activity, ROS production, the total protein, and MDA level was higher at 12 to 48 hpe with different concentrations of PA. Phagocytic activity increased at 6-12 hpe to 40-80 mg/L of PA, but decreased at 24-48 hpe. The expressions of genes PrxV, CLT-6, MT and BD down-regulated at 3 hpe were observed, while differential expressions from 6-48 hpe with different concentrations of PA. The present study demonstrated that immersing A. irradians in PA at effective concentrations could result in differential effects on non-specific immune responses and expressions of immune-related genes. PMID:27171074

  7. Marine Toxin Okadaic Acid Affects the Immune Function of Bay Scallop (Argopecten irradians).

    PubMed

    Chi, Cheng; Giri, Sib Sankar; Jun, Jin Woo; Kim, Hyoun Joong; Yun, Saekil; Kim, Sang Guen; Park, Se Chang

    2016-01-01

    Okadaic acid (OA) is produced by dinoflagellates during harmful algal blooms and is a diarrhetic shellfish poisoning toxin. This toxin is particularly problematic for bivalves that are cultured for human consumption. This study aimed to reveal the effects of exposure to OA on the immune responses of bay scallop, Argopecten irradians. Various immunological parameters were assessed (total hemocyte counts (THC), reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), lactate dehydrogenase (LDH), and nitric oxide (NO) in the hemolymph of scallops at 3, 6, 12, 24, and 48 h post-exposure (hpe) to different concentrations of OA (50, 100, and 500 nM). Moreover, the expression of immune-system-related genes (CLT-6, FREP, HSP90, MT, and Cu/ZnSOD) was also measured. Results showed that ROS, MDA, and NO levels and LDH activity were enhanced after exposure to different concentrations of OA; however, both THC and GSH decreased between 24-48 hpe. The expression of immune-system-related genes was also assessed at different time points during the exposure period. Overall, our results suggest that exposure to OA had negative effects on immune system function, increased oxygenic stress, and disrupted metabolism of bay scallops. PMID:27563864

  8. Purple sea urchin Strongylocentrotus purpuratus gamete manipulation using optical trapping and microfluidics

    NASA Astrophysics Data System (ADS)

    Chandsawangbhuwana, Charlie; Shi, Linda Z.; Zhu, Qingyuan; Berns, Michael W.

    2013-04-01

    A system has been developed that allows for optical and fluidic manipulation of gametes. The optical manipulation is performed by using a single-point gradient trap with a 40× oil immersion PH3 1.3 NA objective on a Zeiss inverted microscope. The fluidic manipulation is performed by using a custom microfluidic chamber designed to fit into the short working distance between the condenser and objective. The system is validated using purple sea urchin Strongylocentrotus purpuratus gametes and has the potential to be used for mammalian in vitro fertilization and animal husbandry.

  9. A STAGE-BASED POPULATION MODEL FOR BAY SCALLOPS (ARGOPECTEN IRRADIANS) AND IMPLICATIONS FOR POPULATION-LEVEL EFFECTS OF HABITAT ATLERATION.

    EPA Science Inventory

    Bay scallops (Argopecten irradians) inhabit shallow subtidal habitats along the Atlantic coast of the United States and require settlement substrates, such as submerged aquatic vegetation (SAV), for their early juvenile stages. The short lifespan of bay scallops (1-2 yr) coupled...

  10. Environmental economics reality check: a case study of the Abanico Medicinal Plant and Organic Agriculture Microenterprise Project.

    PubMed

    Isla, Ana; Thompson, Shirley

    2003-01-01

    This paper presents a case study of the Abanico Medicinal Plant and Organic Agriculture Microenterprise Project in the Arenal Conservation Area, Costa Rica. Microenterprise is the Sustainable Development and the Women in Development model for gender equity and environment of the World Bank, International Monetary Fund and large non-government organizations, like the World Wildlife Fund-Canada. The authors of this paper argue that debt-for-nature investment in microenterprise and ecological economic models are not distinct from neoclassical economic and development models that created the environmental, social and cultural crises in the first place. This case study shows that the world market accommodates only one model of development: unsustainable export-oriented production based on flexible labour markets, low wages, indebtedness and low cost production. Working standards in those micro-enterprises are eroded due to many factors,including indebtedness. What happened at a national level in non-industrial countries with the international debt crisis is now mirrored in individual indebtedness through microenterprise. Is current development policy creating a new form of indentured servitude? Medicinal plants, prior to commodification, were a source of women's power and upon commodification in international development projects, are the source of their exploitation. PMID:12859005

  11. Gene structure in the sea urchin Strongylocentrotus purpuratus based on transcriptome analysis.

    PubMed

    Tu, Qiang; Cameron, R Andrew; Worley, Kim C; Gibbs, Richard A; Davidson, Eric H

    2012-10-01

    A comprehensive transcriptome analysis has been performed on protein-coding RNAs of Strongylocentrotus purpuratus, including 10 different embryonic stages, six feeding larval and metamorphosed juvenile stages, and six adult tissues. In this study, we pooled the transcriptomes from all of these sources and focused on the insights they provide for gene structure in the genome of this recently sequenced model system. The genome had initially been annotated by use of computational gene model prediction algorithms. A large fraction of these predicted genes were recovered in the transcriptome when the reads were mapped to the genome and appropriately filtered and analyzed. However, in a manually curated subset, we discovered that more than half the computational gene model predictions were imperfect, containing errors such as missing exons, prediction of nonexistent exons, erroneous intron/exon boundaries, fusion of adjacent genes, and prediction of multiple genes from single genes. The transcriptome data have been used to provide a systematic upgrade of the gene model predictions throughout the genome, very greatly improving the research usability of the genomic sequence. We have constructed new public databases that incorporate information from the transcriptome analyses. The transcript-based gene model data were used to define average structural parameters for S. purpuratus protein-coding genes. In addition, we constructed a custom sea urchin gene ontology, and assigned about 7000 different annotated transcripts to 24 functional classes. Strong correlations became evident between given functional ontology classes and structural properties, including gene size, exon number, and exon and intron size. PMID:22709795

  12. Development of marine sediment toxicity identification evaluation methods using Strongylocentrotus purpuratus, Mytilus edulis, and Eohaustorius estuarius

    SciTech Connect

    Wortham, G.; Cotsifas, J.S.; Taberski, K.; Hansen, S.R.

    1994-12-31

    Widespread sediment toxicity, including ``clean`` reference sites, dictates that the causes of toxicity in sediments be determined. Toxicity Identification Evaluations (TIE) are useful tools in characterizing compounds responsible for toxicity, but were unavailable for sediment samples. TIE methods were developed for sediment porewater and included the following components: determination of an appropriate porewater extraction process; control TIE tests using marine water and porewater evaluating species sensitivities to the fractionation procedures; validation experiments investigating the removal efficiencies of organics using C18 solid phase extraction, and metals chelation using EDTA and STS; spiking experiments to determine the effectiveness of the TIE procedure in identifying multiple toxicants. The authors determined that fractionation procedures could be applied to both marine water and porewater using S. purpuratus, M. edulis and E. estuarius as biological detectors.

  13. Developmental cis-regulatory analysis of the cyclin D gene in the sea urchin Strongylocentrotus purpuratus

    PubMed Central

    McCarty, Christopher M.

    2013-01-01

    Cyclin D genes regulate the cell cycle, growth and differentiation in response to intercellular signaling. While the promoters of vertebrate cyclin D genes have been analyzed, the cis-regulatory sequences across an entire cyclin D locus have not. Doing so would increase understanding of how cyclin D genes respond to the regulatory states established by developmental gene regulatory networks, linking cell cycle and growth control to the ontogenetic program. Therefore, we conducted a cis-regulatory analysis on the cyclin D gene, SpcycD, of the sea urchin, Strongylocentrotus purpuratus, during embryogenesis, identifying upstream and intronic sequences, located within six defined regions bearing one or more cis-regulatory modules each. PMID:24090975

  14. Genomic in situ hybridization identifies parental chromosomes in hybrid scallop (Bivalvia, Pectinoida, Pectinidae) between female Chlamys farreri and male Argopecten irradians irradians

    PubMed Central

    Huang, Xiaoting; Bi, Ke; Lu, Wei; Wang, Shi; Zhang, Lingling; Bao, Zhenmin

    2015-01-01

    Abstract Interspecific crossing was artificially carried out between Chlamys farreri (Jones & Preston, 1904) ♀ and Argopecten irradians irradians (Lamarck, 1819) ♂, two of the dominant cultivated scallop species in China. Genomic in situ hybridization (GISH) was used to examine the chromosome constitution and variation in hybrids at early embryonic stage. The number of chromosomes in 66.38% of the metaphases was 2n = 35 and the karyotype was 2n = 3 m + 5 sm + 16 st + 11 t. After GISH, two parental genomes were clearly distinguished in hybrids, most of which comprised 19 chromosomes derived from their female parent (Chlamys farreri) and 16 chromosomes from their male parent (Argopecten irradians irradians). Some chromosome elimination and fragmentation was also observed in the hybrids. PMID:26140161

  15. DspA from Strongylocentrotus purpuratus: The first biochemically characterized haloalkane dehalogenase of non-microbial origin.

    PubMed

    Fortova, Andrea; Sebestova, Eva; Stepankova, Veronika; Koudelakova, Tana; Palkova, Lenka; Damborsky, Jiri; Chaloupkova, Radka

    2013-11-01

    Haloalkane dehalogenases are known as bacterial enzymes cleaving a carbon-halogen bond in halogenated compounds. Here we report the first biochemically characterized non-microbial haloalkane dehalogenase DspA from Strongylocentrotus purpuratus. The enzyme shows a preference for terminally brominated hydrocarbons and enantioselectivity towards β-brominated alkanes. Moreover, we identified other putative haloalkane dehalogenases of eukaryotic origin, representing targets for future experiments to discover dehalogenases with novel catalytic properties. PMID:23939220

  16. Ocean acidification research in the 'post-genomic' era: Roadmaps from the purple sea urchin Strongylocentrotus purpuratus.

    PubMed

    Evans, Tyler G; Padilla-Gamiño, Jacqueline L; Kelly, Morgan W; Pespeni, Melissa H; Chan, Francis; Menge, Bruce A; Gaylord, Brian; Hill, Tessa M; Russell, Ann D; Palumbi, Stephen R; Sanford, Eric; Hofmann, Gretchen E

    2015-07-01

    Advances in nucleic acid sequencing technology are removing obstacles that historically prevented use of genomics within ocean change biology. As one of the first marine calcifiers to have its genome sequenced, purple sea urchins (Strongylocentrotus purpuratus) have been the subject of early research exploring genomic responses to ocean acidification, work that points to future experiments and illustrates the value of expanding genomic resources to other marine organisms in this new 'post-genomic' era. This review presents case studies of S. purpuratus demonstrating the ability of genomic experiments to address major knowledge gaps within ocean acidification. Ocean acidification research has focused largely on species vulnerability, and studies exploring mechanistic bases of tolerance toward low pH seawater are comparatively few. Transcriptomic responses to high pCO₂ seawater in a population of urchins already encountering low pH conditions have cast light on traits required for success in future oceans. Secondly, there is relatively little information on whether marine organisms possess the capacity to adapt to oceans progressively decreasing in pH. Genomics offers powerful methods to investigate evolutionary responses to ocean acidification and recent work in S. purpuratus has identified genes under selection in acidified seawater. Finally, relatively few ocean acidification experiments investigate how shifts in seawater pH combine with other environmental factors to influence organism performance. In S. purpuratus, transcriptomics has provided insight into physiological responses of urchins exposed simultaneously to warmer and more acidic seawater. Collectively, these data support that similar breakthroughs will occur as genomic resources are developed for other marine species. PMID:25773301

  17. Phosphoproteomic network analysis in the sea urchin Strongylocentrotus purpuratus reveals new candidates in egg activation.

    PubMed

    Guo, Hongbo; Garcia-Vedrenne, Ana Elisa; Isserlin, Ruth; Lugowski, Andrew; Morada, Anthony; Sun, Alex; Miao, Yishen; Kuzmanov, Uros; Wan, Cuihong; Ma, Hongyue; Foltz, Kathy; Emili, Andrew

    2015-12-01

    Fertilization triggers a dynamic symphony of molecular transformations induced by a rapid rise in intracellular calcium. Most prominent are surface alterations, metabolic activation, cytoskeletal reorganization, and cell-cycle reentry. While the activation process appears to be broadly evolutionarily conserved, and protein phosphorylation is known to play a key role, the signaling networks mediating the response to fertilization are not well described. To address this gap, we performed a time course phosphoproteomic analysis of egg activation in the sea urchin Strongylocentrotus purpuratus, a system that offers biochemical tractability coupled with exquisite synchronicity. By coupling large-scale phosphopeptide enrichment with unbiased quantitative MS, we identified striking changes in global phosphoprotein patterns at 2- and 5-min postfertilization as compared to unfertilized eggs. Overall, we mapped 8796 distinct phosphosite modifications on 2833 phosphoproteins, of which 15% were differentially regulated in early egg activation. Activated kinases were identified by phosphosite mapping, while enrichment analyses revealed conserved signaling cascades not previously associated with egg activation. This work represents the most comprehensive study of signaling associated with egg activation to date, suggesting novel mechanisms that can be experimentally tested and providing a valuable resource for the broader research community. All MS data have been deposited in the ProteomeXchange with identifier PXD002239 (http://proteomecentral.proteomexchange.org/dataset/PXD002239). PMID:26227301

  18. Identification and developmental expression of the ets gene family in the sea urchin (Strongylocentrotus purpuratus).

    PubMed

    Rizzo, Francesca; Fernandez-Serra, Montserrat; Squarzoni, Paola; Archimandritis, Aristea; Arnone, Maria I

    2006-12-01

    A systematic search in the available scaffolds of the Strongylocentrotus purpuratus genome has revealed that this sea urchin has 11 members of the ets gene family. A phylogenetic analysis of these genes showed that almost all vertebrate ets subfamilies, with the exception of one, so far found only in mammals, are each represented by one orthologous sea urchin gene. The temporal and spatial expression of the identified ETS factors was also analyzed during embryogenesis. Five ets genes (Sp-Ets1/2, Sp-Tel, Sp-Pea, Sp-Ets4, Sp-Erf) are also maternally expressed. Three genes (Sp-Elk, Sp-Elf, Sp-Erf) are ubiquitously expressed during embryogenesis, while two others (Sp-Gabp, Sp-Pu.1) are not transcribed until late larval stages. Remarkably, five of the nine sea urchin ets genes expressed during embryogenesis are exclusively (Sp-Ets1/2, Sp-Erg, Sp-Ese) or additionally (Sp-Tel, Sp-Pea) expressed in mesenchyme cells and/or their progenitors. Functional analysis of Sp-Ets1/2 has previously demonstrated an essential role of this gene in the specification of the skeletogenic mesenchyme lineage. The dynamic, and in some cases overlapping and/or unique, developmental expression pattern of the latter five genes suggests a complex, non-redundant function for ETS factors in sea urchin mesenchyme formation and differentiation. PMID:16997294

  19. The chemical defensome: Environmental sensing and response genes in the Strongylocentrotus purpuratus genome

    PubMed Central

    Goldstone, J.V.; Hamdoun, A.; Cole, B.J.; Howard-Ashby, M.; Nebert, D.W.; Scally, M.; Dean, M.; Epel, D.; Hahn, M.E.; Stegeman, J.J.

    2011-01-01

    Metazoan genomes contain large numbers of genes that participate in responses to environmental stressors. We surveyed the sea urchin Strongylocentrotus purpuratus genome for homologs of gene families thought to protect against chemical stressors; these genes collectively comprise the ‘chemical defensome.’ Chemical defense genes include cytochromes P450 and other oxidases, various conjugating enzymes, ATP-dependent efflux transporters, oxidative detoxification proteins, and transcription factors that regulate these genes. Together such genes account for more than 400 genes in the sea urchin genome. The transcription factors include homologs of the aryl hydrocarbon receptor, hypoxia-inducible factor, nuclear factor erythroid-derived 2, heat shock factor, and nuclear hormone receptors, which regulate stress-response genes in vertebrates. Some defense gene families, including the ABCC, the UGT, and the CYP families, have undergone expansion in the urchin relative to other deuterostome genomes, whereas the stress sensor gene families do not show such expansion. More than half of the defense genes are expressed during embryonic or larval life stages, indicating their importance during development. This genome-wide survey of chemical defense genes in the sea urchin reveals evolutionary conservation of this network combined with lineage-specific diversification that together suggest the importance of these chemical stress sensing and response mechanisms in early deuterostomes. These results should facilitate future studies on the evolution of chemical defense gene networks and the role of these networks in protecting embryos from chemical stress during development. PMID:17097629

  20. Genetic analysis of the purplish Washington clam (Saxidomus purpuratus Sowerby) of Korean coastal waters.

    PubMed

    Cho, Eun-Seob; Seo, Young-Il; Suh, Young-Sang

    2013-05-01

    To investigate the genetic structure of the purplish Washington clam population, Saxidomus purpuratus Sowerby, in Korea. A portion of mitochondrial COI gene sequences (605 bp) for phylogenetic comparison was determined. Sequence analysis of 62 individuals collected from six regions revealed 13 haplotypes. Phylogenetic analysis using Phylogeny Inference Package (PHYLIP) subdivided the purplish Washington clam into two clades (termed clade A and B), weak supported groups (< 65 of bootstrap value). This haplotype subdivision was also in accordance with geographic separation; one each at Masan, Yeosu, Samcheonpo, Jubyeon and Geojedo, and the other at Sineju. Population genetic analysis subdivided these two population groups with a geographic distance (d = 0.431, p = 0.379). Furthermore, in the Sineju population, the maximum sequence divergence (2.67%) and minimum nucleotide diversity (0.0012426) were shown in which might be reflective of a relatively small population size and the geographical isolation of the population as compared with other populations. However, a very high migration rate (N(m) = 59.62-infinite) and a very low level of geographic distance (F(ST) = -0.076-0.055) were noted to exist among the South and East Sea populations, suggesting that individuals between populations should show a significantly active genetic mixing and migration regardless of geography. These findings allowed us to conclude that the purplish Washington clam populations occurring in the South and East Sea were formed with randomly dispersed individuals. PMID:24617150

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

    PubMed Central

    Harris, Todd R.; Aronov, Pavel A.

    2008-01-01

    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. PMID:18554159

  2. Spatial and temporal variation in results of purple urchin (Strongylocentrotus purpuratus) toxicity tests with zinc

    SciTech Connect

    Phillips, B.M.; Anderson, B.S.; Hunt, J.W.

    1998-03-01

    Purple urchins (Strongylocentrotus purpuratus) were maintained in year-round spawning condition in the laboratory for use in fertilization and larval development experiments designed to assess temporal variability in response to zinc. Results of these tests were compared to those from tests using gametes obtained from a field-collected population. Fertilization and larval development tests were also conducted comparing field-collected purple urchins from three geographically distinct groups on the West Coast of the United States. Fertilization tests conducted to assess temporal variability produced variable median effects concentrations (EC50s) ranging from 4.1 to >100 {micro}g/L zinc. Larval development tests did not demonstrate significant differences in response to zinc between geographically distinct purple urchin populations. Fertilization test variability was examined in terms of sperm concentration and sperm collection method during two seasons. Reduced variability was found with dry sperm collection in tests conducted in March 1995 but increased again in tests conducted in June 1995, regardless of sperm collection method. Increased variability in response to zinc may be caused by seasonal temperature effects.

  3. Characterization of the proteins comprising the integral matrix of Strongylocentrotus purpuratus embryonic spicules

    NASA Technical Reports Server (NTRS)

    Killian, C. E.; Wilt, F. H.

    1996-01-01

    In the present study, we enumerate and characterize the proteins that comprise the integral spicule matrix of the Strongylocentrotus purpuratus embryo. Two-dimensional gel electrophoresis of [35S]methionine radiolabeled spicule matrix proteins reveals that there are 12 strongly radiolabeled spicule matrix proteins and approximately three dozen less strongly radiolabeled spicule matrix proteins. The majority of the proteins have acidic isoelectric points; however, there are several spicule matrix proteins that have more alkaline isoelectric points. Western blotting analysis indicates that SM50 is the spicule matrix protein with the most alkaline isoelectric point. In addition, two distinct SM30 proteins are identified in embryonic spicules, and they have apparent molecular masses of approximately 43 and 46 kDa. Comparisons between embryonic spicule matrix proteins and adult spine integral matrix proteins suggest that the embryonic 43-kDa SM30 protein is an embryonic isoform of SM30. An adult 49-kDa spine matrix protein is also identified as a possible adult isoform of SM30. Analysis of the SM30 amino acid sequences indicates that a portion of SM30 proteins is very similar to the carbohydrate recognition domain of C-type lectin proteins.

  4. Cell surface of sea urchin micromeres and primary mesenchyme. [Arbacia punctulata; Strongylocentrotus drobachiensis; Strongylocentrotus purpuratus

    SciTech Connect

    DeSimone, D.W.

    1985-01-01

    The cell surface and extracellular matrix (ECM) of the sea urchin embryo were studied during the early morphogenetic events involved in the differentiation of the micromere cell lineage. Sixteen-cell and early cleavage stage blastomeres were isolated and the protein composition of their cell surfaces examined by /sup 125/I-labelling followed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Micromere-specific cell surface proteins are reported for Arbacia punctulata, Strongylocentrotus droebachiensis, and Strongylocentrotus purpuratus. Cell surface glycoproteins were characterized on the basis of lectin binding specificity with a novel lectin affinity transfer technique. Using this procedure, cell-type specific surface proteins, which are also lectin-binding specific, can be detected. In addition, fluorescein conjugated lectins were microinjected into the blastocoels of living S. drobachiensis and Lytechinus pictus embryos and the patterns of lectin bindings observed by fluorescence microscopy. The evidence presented in this thesis suggests that the differentiation of the primary mesenchyme cells is correlated with changes in the molecular composition of the cell-surface and the ECM.

  5. Geographical variation of shell thickness in the mussel Perumytilus purpuratus along the southeast Pacific coast.

    PubMed

    Briones, Carolina; Rivadeneira, Marcelo M; Fernández, Miriam; Guiñez, Ricardo

    2014-12-01

    At broad geographical scales, the variation in bivalve shell thickness can be modulated by environmental factors that vary with latitude, such as sea surface temperature (SST), seawater pH, or calcium carbonate availability. Mussels usually form multilayered beds, and shell thickness is also expected to be affected by density and layering due to intraspecific competition. In this work, we explored the geographical variation of shell thickness in the intertidal mussel Perumytilus purpuratus between 18° and 42°S along the southeastern Pacific coast. We tested the hypothesis that there was a positive relationship between shell thickness and SST, and then we explored other variables that could have an effect on thickness, such as density, number of layers, and others environmental variables (pH and calcite concentration). The expected positive linear relationship between shell thickness and sea surface temperature was not found, but when the other population variables were included in the analysis, an unexpected inverse SST-thickness relationships appeared as significant, probably because this species could be adapted to colder and more acid seawater as are those of the tips of South America. Thickness was also negatively affected by density, which was expected for a gregarious species showing high intraspecific competition. Finally, our results highlight the importance of including density and crowding effects when macroscale patterns are explored, particularly in gregarious species, since these patterns could also be modulated by density-dependent processes, which might then override latitudinal trends of shell thickness when they are not included in the analyses. PMID:25572210

  6. Elimination of 2,4,5,2',4',5'-hexachlorobiphenyl by the purple sea urchin, Strongylocentrotus purpuratus, following single exposure

    SciTech Connect

    Tjeerdema, R.S.; Jacobs, R.S.

    1987-06-01

    Understanding the fate of a single PCB isomer in a resident species may aid in assessing the risk to the marine community. Therefore, the elimination of 2,4,5,2',4',5'-hexachlorobiphenyl (HCBP) by the purple sea urchin, Strongylocentrotus purpuratus, following a single exposure, was investigated. The purple sea urchin was chosen because of its economic importance and ability to proliferate in certain polluted conditions. Single exposure may best mimic the effects of intermittent oceanic incineration or disposal, and 2,4,5,2',4',5'-HCBP was chosen due to its presence in common PCB mixtures and high chlorine content, thus strong lipophilicity.

  7. Manipulation of Developing Juvenile Structures in Purple Sea Urchins (Strongylocentrotus purpuratus) by Morpholino Injection into Late Stage Larvae

    PubMed Central

    2014-01-01

    Sea urchins have been used as experimental organisms for developmental biology for over a century. Yet, as is the case for many other marine invertebrates, understanding the development of the juveniles and adults has lagged far behind that of their embryos and larvae. The reasons for this are, in large part, due to the difficulty of experimentally manipulating juvenile development. Here we develop and validate a technique for injecting compounds into juvenile rudiments of the purple sea urchin, Strongylocentrotus purpuratus. We first document the distribution of rhodaminated dextran injected into different compartments of the juvenile rudiment of sea urchin larvae. Then, to test the potential of this technique to manipulate development, we injected Vivo-Morpholinos (vMOs) designed to knock down p58b and p16, two proteins involved in the elongation of S. purpuratus larval skeleton. Rudiments injected with these vMOs showed a delay in the growth of some juvenile skeletal elements relative to controls. These data provide the first evidence that vMOs, which are designed to cross cell membranes, can be used to transiently manipulate gene function in later developmental stages in sea urchins. We therefore propose that injection of vMOs into juvenile rudiments, as shown here, is a viable approach to testing hypotheses about gene function during development, including metamorphosis. PMID:25436992

  8. Geographical Variations and Genetic Distances of Three Saxidomus purpuratus Populations ascertained by PCR Analysis.

    PubMed

    Yoon, Jong-Man

    2015-12-01

    Genomic DNA samples isolated from geographical purplish Washington clam (Saxidomus purpuratus) were obtained from three different regions in the Korean Peninsula: Geoje (Geoje population; GJP), Gunsan (Gunsan population; GSP) and a site of North Korea (North Korea population; NKP). The seven primers generated the total 369 loci that can be scored from the GSP clam population. 356 fragments were generated from the NKP clam population. The complexity of the banding patterns varies dramatically between the primers and three localities. In this study, 319 loci were identified in the purplish Washington clam from Geoje and 369 in the clam population from Gunsan: 221 specific loci (69.3%) in the GJP clam population and 300 (81.3%) in the GSP population. These results demonstrate that the primer detected a large quantity of specific fragments, suggesting that the genetic variation in the GSP is higher than in the GJP population. In particular, the BION-28 primer gave DNA profiles with more fragments than the other six primers in the NKP population. The oligonucleotides primer BION-75 produced 21 unique loci to each population, which were ascertaining each population, approximately 250 bp, 300 bp and 400 bp, in the GJP population. Outstandingly, the primer BION-50 detected 21 shared loci by the three populations, major and/or minor fragments of sizes 150 bp, which were matching in all samples. With regard to average bandsharing value (BS) results, individuals from GJP population (0.743) displayed higher bandsharing values than did individuals from GSP population (0.606). In the present study, the dendrogram gained by the seven oligonucleotides primers indicates three genetic clusters: cluster 1 (GEOJE 01 ~ GEOJE 07), cluster 2 (GUNSAN 08 ~ GUNSAN 14), cluster 3 (N.KOREA 15 ~ N.KOREA 21). Among the twenty one clams, the shortest genetic distance that revealed significant molecular differences was between individuals 08 and 09 from the NKP population (genetic distance = 0

  9. Early exposure of bay scallops (Argopecten irradians) to high CO₂ causes a decrease in larval shell growth.

    PubMed

    White, Meredith M; McCorkle, Daniel C; Mullineaux, Lauren S; Cohen, Anne L

    2013-01-01

    Ocean acidification, characterized by elevated pCO₂ and the associated decreases in seawater pH and calcium carbonate saturation state (Ω), has a variable impact on the growth and survival of marine invertebrates. Larval stages are thought to be particularly vulnerable to environmental stressors, and negative impacts of ocean acidification have been seen on fertilization as well as on embryonic, larval, and juvenile development and growth of bivalve molluscs. We investigated the effects of high CO₂ exposure (resulting in pH = 7.39, Ω(ar) = 0.74) on the larvae of the bay scallop Argopecten irradians from 12 h to 7 d old, including a switch from high CO₂ to ambient CO₂ conditions (pH = 7.93, Ω(ar) = 2.26) after 3 d, to assess the possibility of persistent effects of early exposure. The survival of larvae in the high CO₂ treatment was consistently lower than the survival of larvae in ambient conditions, and was already significantly lower at 1 d. Likewise, the shell length of larvae in the high CO₂ treatment was significantly smaller than larvae in the ambient conditions throughout the experiment and by 7 d, was reduced by 11.5%. This study also demonstrates that the size effects of short-term exposure to high CO₂ are still detectable after 7 d of larval development; the shells of larvae exposed to high CO₂ for the first 3 d of development and subsequently exposed to ambientCO₂ were not significantly different in size at 3 and 7 d than the shells of larvae exposed to high CO₂ throughout the experiment. PMID:23596514

  10. Early Exposure of Bay Scallops (Argopecten irradians) to High CO2 Causes a Decrease in Larval Shell Growth

    PubMed Central

    White, Meredith M.; McCorkle, Daniel C.; Mullineaux, Lauren S.; Cohen, Anne L.

    2013-01-01

    Ocean acidification, characterized by elevated pCO2 and the associated decreases in seawater pH and calcium carbonate saturation state (Ω), has a variable impact on the growth and survival of marine invertebrates. Larval stages are thought to be particularly vulnerable to environmental stressors, and negative impacts of ocean acidification have been seen on fertilization as well as on embryonic, larval, and juvenile development and growth of bivalve molluscs. We investigated the effects of high CO2 exposure (resulting in pH = 7.39, Ωar = 0.74) on the larvae of the bay scallop Argopecten irradians from 12 h to 7 d old, including a switch from high CO2 to ambient CO2 conditions (pH = 7.93, Ωar = 2.26) after 3 d, to assess the possibility of persistent effects of early exposure. The survival of larvae in the high CO2 treatment was consistently lower than the survival of larvae in ambient conditions, and was already significantly lower at 1 d. Likewise, the shell length of larvae in the high CO2 treatment was significantly smaller than larvae in the ambient conditions throughout the experiment and by 7 d, was reduced by 11.5%. This study also demonstrates that the size effects of short-term exposure to high CO2 are still detectable after 7 d of larval development; the shells of larvae exposed to high CO2 for the first 3 d of development and subsequently exposed to ambient CO2 were not significantly different in size at 3 and 7 d than the shells of larvae exposed to high CO2 throughout the experiment. PMID:23596514

  11. What is the core oscillator in the speract-activated pathway of the Strongylocentrotus purpuratus sperm flagellum?

    PubMed

    Aguilera, Luis U; Galindo, Blanca E; Sánchez, Daniel; Santillán, Moisés

    2012-06-01

    Sperm chemotaxis has an important role in fertilization. Most of our knowledge regarding this phenomenon comes from studies in organisms whose fertilization occurs externally, like sea urchins. Sea urchin spermatozoa respond to sperm-activating peptides, which diffuse from the egg jelly coat and interact with their receptor in the flagellum, triggering several physiological responses: changes in membrane potential, intracellular pH, cyclic nucleotide levels, and intracellular Ca2+ concentration ([Ca2+]). In particular, flagellar [Ca2+] has been shown to oscillate. These [Ca2+] oscillations are correlated with changes in the flagellar shape and so with the regulation of the sperm swimming paths. In this study, we demonstrate, from a mathematical modeling perspective, that the reported speract-activated signaling pathway in Strongylocentrotus purpuratus (speract being a sperm-activating peptide specific to this species) has the necessary elements to replicate the reported [Ca2+] oscillations. We further investigate which elements of this signaling pathway constitute the core oscillator. PMID:22713563

  12. Phosphoproteomes of Strongylocentrotus purpuratus shell and tooth matrix: identification of a major acidic sea urchin tooth phosphoprotein, phosphodontin

    PubMed Central

    2010-01-01

    Background Sea urchin is a major model organism for developmental biology and biomineralization research. However, identification of proteins involved in larval skeleton formation and mineralization processes in the embryo and adult, and the molecular characterization of such proteins, has just gained momentum with the sequencing of the Strongylocentrotus purpuratus genome and the introduction of high-throughput proteomics into the field. Results The present report contains the determination of test (shell) and tooth organic matrix phosphoproteomes. Altogether 34 phosphoproteins were identified in the biomineral organic matrices. Most phosphoproteins were specific for one compartment, only two were identified in both matrices. The sea urchin phosphoproteomes contained several obvious orthologs of mammalian proteins, such as a Src family tyrosine kinase, protein kinase C-delta 1, Dickkopf-1 and other signal transduction components, or nucleobindin. In most cases phosphorylation sites were conserved between sea urchin and mammalian proteins. However, the majority of phosphoproteins had no mammalian counterpart. The most interesting of the sea urchin-specific phosphoproteins, from the perspective of biomineralization research, was an abundant highly phosphorylated and very acidic tooth matrix protein composed of 35 very similar short sequence repeats, a predicted N-terminal secretion signal sequence, and an Asp-rich C-terminal motif, contained in [Glean3:18919]. Conclusions The 64 phosphorylation sites determined represent the most comprehensive list of experimentally identified sea urchin protein phosphorylation sites at present and are an important addition to the recently analyzed Strongylocentrotus purpuratus shell and tooth proteomes. The identified phosphoproteins included a major, highly phosphorylated protein, [Glean3:18919], for which we suggest the name phosphodontin. Although not sequence-related to such highly phosphorylated acidic mammalian dental

  13. Responses to two-way selection on growth in mass-spawned F1 progeny of Argopecten irradians concentricus (Say)

    NASA Astrophysics Data System (ADS)

    Wang, Hui; Liu, Jin; Li, Yanhong; Zhu, Xiaowen; Liu, Zhigang

    2014-03-01

    In the present study, the effect of one-generation divergent selection on the growth and survival of the bay scallop ( Argopecten irradians concentricus) was examined to evaluate the efficacy of a selection program currently being carried out in Beibu Bay in the South China Sea. A total of 146 adult scallops were randomly selected from the same cultured population of A. i. concentricus, and divided into two groups in shell length (anterior-posterior measurement): large (4.91-6.02 cm, n=74) and small (3.31-4.18 cm, n=72). At the same time, a control group was also randomly sampled (4.21-4.88 cm, n =80). Mass-spawned F 1 progenies from the three size groups were obtained and reared under identical conditions at all growth phases. The effects of two-way (or upward-downward) selection on fertilization rate, hatching rate, survival rate, daily growth in shell length and body weight were assessed in the three size groups. Results show that significant differences ( P<0.01) were found in hatching rate, survival rate and daily growth of F1 progenies, but not in fertilization rate ( P>0.05), among the three groups. The hatching rate, survival rate and daily growth of the progeny of large-sized parents were greater than those of the control group ( P<0.05), which in turn were larger than those of small-sized group ( P<0.05). Responses to selection by shell length and body weight were 0.32 ± 0.04 cm and 2.18 ± 0.05 g, respectively, for the upward selection, and -0.14 ± 0.03 cm and -2.77 ± 0.06 g, respectively, for the downward selection. The realized heritability estimates of shell length and body weight were 0.38 ± 0.06 cm and 0.22 ± 0.07 g for the upward selection, and 0.24 ± 0.06 cm and 0.37 ± 0.09 g for the downward selection, respectively. The change in growth by bidirectional selection suggests that high genetic variation may be present in the cultured bay scallop population in China.

  14. Developmental expression of a cell surface protein involved in sea urchin skeleton formation. [Strongylocentrotus purpuratus; Lytechinus pictus

    SciTech Connect

    Farach, M.C.; Valdizan, M.; Park, H.R.; Decker, G.L.; Lennarz, W.J.

    1986-05-01

    The authors have previously used a monoclonal antibody (1223) to identify a 130 Kd cell surface protein involved in skeleton formation is sea urchin embryos. In the current study the authors have examined the expression of the 1223 antigen over the course of development of embryos of two species, Strongylocentrotus purpuratus and Lytechinus pictus. The 130 Kd protein is detected in S. purp eggs on immunoblots. Labeling with (/sup 3/H) leucine and immunoaffinity chromatography show that it also is synthesized shortly after fertilization. Immunofluroescence reveals that at this early stage the 1223 antigen is uniformly distributed on all of the cells. Synthesis decreases to a minimum by the time of hatching (18 h), as does the total amount of antigen present in the embryo. A second period of synthesis commences at the mesenchyme blastula stage, when the spicule-forming primary mesenchyme cells (PMCs) have appeared. During this later stage, synthesis and cell surface expression are restricted to the PMCs. In contrast to S. purp., in L. pictus the 130 Kd protein does not appear until the PMCs are formed. Hybrid embryos demonstrate a pattern of expression of the maternal species. These results suggest that early expression of 1223 antigen in S. purp. is due to utilization of maternal transcripts present in the egg. In both species later expression in PMCs appears to be the result of cell-type specific synthesis, perhaps encoded by embryonic transcripts.

  15. Protein tyrosine and serine–threonine phosphatases in the sea urchin, Strongylocentrotus purpuratus: Identification and potential functions

    PubMed Central

    Byrum, C.A.; Walton, K.D.; Robertson, A.J.; Carbonneau, S.; Thomason, R.T.; Coffman, J.A.; McClay, D.R.

    2011-01-01

    Protein phosphatases, in coordination with protein kinases, play crucial roles in regulation of signaling pathways. To identify protein tyrosine phosphatases (PTPs) and serine–threonine (ser–thr) phosphatases in the Strongylocentrotus purpuratus genome, 179 annotated sequences were studied (122 PTPs, 57 ser–thr phosphatases). Sequence analysis identified 91 phosphatases (33 conventional PTPs, 31 dual specificity phosphatases, 1 Class III Cysteine-based PTP, 1 Asp-based PTP, and 25 ser–thr phosphatases). Using catalytic sites, levels of conservation and constraint in amino acid sequence were examined. Nine of 25 receptor PTPs (RPTPs) corresponded to human, nematode, or fly homologues. Domain structure revealed that sea urchin-specific RPTPs including two, PTPRLec and PTPRscav, may act in immune defense. Embryonic transcription of each phosphatase was recorded from a high-density oligonucleotide tiling microarray experiment. Most RPTPs are expressed at very low levels, whereas nonreceptor PTPs (NRPTPs) are generally expressed at moderate levels. High expression was detected in MAP kinase phosphatases (MKPs) and numerous ser–thr phosphatases. For several expressed NRPTPs, MKPs, and ser–thr phosphatases, morpholino antisense-mediated knockdowns were performed and phenotypes obtained. Finally, to assess roles of annotated phosphatases in endomesoderm formation, a literature review of phosphatase functions in model organisms was superimposed on sea urchin developmental pathways to predict areas of functional activity. PMID:17087928

  16. Effects of increased pCO2 and geographic origin on purple sea urchin (Strongylocentrotus purpuratus) calcite elemental composition

    NASA Astrophysics Data System (ADS)

    LaVigne, M.; Hill, T. M.; Sanford, E.; Gaylord, B.; Russell, A. D.; Lenz, E. A.; Hosfelt, J. D.; Young, M. K.

    2012-12-01

    Ocean acidification will likely have negative impacts on invertebrates producing skeletons composed of calcium carbonate. Skeletal solubility is partly controlled by the incorporation of "foreign" ions (such as Mg and Sr) into the crystal lattice of these skeletal structures, a process that is sensitive to a variety of biological and environmental factors. Here we explore the effects of life stage, oceanographic region of origin, and changes in the partial pressure of carbon dioxide in seawater (pCO2) on trace elemental composition in the purple sea urchin (Strongylocentrotus purpuratus). We show that, similar to other urchin taxa, adult purple sea urchins have the ability to precipitate skeleton composed of a range of biominerals spanning low to high magnesium calcites. Mg/Ca and Sr/Ca ratios were substantially lower in adult spines compared to adult tests. On the other hand, trace elemental composition was invariant among adults collected from four oceanographically distinct regions along the US west coast (Oregon, Northern California, Central California, and Southern California). Skeletons of newly settled juvenile urchins that originated from adults from the four regions exhibited intermediate Mg/Ca and Sr/Ca between adult spine and test endmembers, indicating that skeleton precipitated during early life stages is more soluble than adult spines and less soluble than adult tests. Mean skeletal Mg/Ca or Sr/Ca of juvenile skeleton did not vary with source region when larvae were reared under present-day, global-average seawater carbonate conditions (400 ppm; pH = 8.02 ± 0.03 1 SD; Ωcalcite = 3.3 ± 0.2 1 SD). However, when reared under elevated CO2 (900 ppm; pH = 7.72 ± 0.03; Ωcalcite = 1.8 ± 0.1), skeletal Sr/Ca in juveniles exhibited increased variance across the four regions. Although larvae from the northern populations (Oregon, Northern California, Central California) did not exhibit differences in Mg or Sr incorporation under elevated CO2 (Sr/Ca = 2

  17. Effects of an oil production effluent on gametogenesis and gamete performance in the purple sea urchin (Strongylocentrotus purpuratus Stimpson)

    SciTech Connect

    Krause, P.R. . Dept. of Biological Sciences)

    1994-07-01

    Adult organisms subjected to chronic discharges from a point source of pollution may exhibit several sublethal responses. One such response is the impairment of gamete production. This may be expressed in the amount and/or quality of gametes produced by adults. In this study the effects of chronic exposure to produced water (an oil production effluent) on the gametogenesis and gamete performance of the purple sea urchin (Strongylocentrotus purpuratus Stimpson) were examined using an in situ caging experiment. Adult purple sea urchins were kept in benthic cages arrayed down-field from a discharging diffuser at 13 sites, with distances ranging from 5 to 1,000 m. Cage exposures were maintained in the field for eight weeks, and each cage held 25 animals. Gametogenesis was examined for each sex by comparing a size-independent measure of relative gonads ass as determined by analysis of covariance. Results showed that there was a significant negative relationship between these estimates of relative gonad mass and distance from the outfall for both sexes, indicating that sea urchins living closer to the outfall produced significantly larger gonads. Gamete performance was measured through a fertilization kinetics bioassay that held the concentration of eggs constant and varied the amount of sperm added. The proportion of eggs fertilized under each sperm concentration was determined and the response fit to a model of fertilizability showed a positive relationship with distance away from the outfall. These findings indicate that although adult sea urchins exposed to a produced water outfall exhibit larger gonads, they suffer a marked decrease in a gamete performance.

  18. Effects of five southern California macroalgal diets on consumption, growth, and gonad weight, in the purple sea urchin Strongylocentrotus purpuratus

    PubMed Central

    Byrnes, Jarrett E.K.; Reed, Daniel C.

    2015-01-01

    Consumer growth and reproductive capacity are direct functions of diet. Strongylocentrotid sea urchins, the dominant herbivores in California kelp forests, strongly prefer giant kelp (Macrocystis pyrifera), but are highly catholic in their ability to consume other species. The biomass of Macrocystis fluctuates greatly in space and time, and the extent to which urchins can use alternate species of algae or a mixed diet of multiple algal species to maintain fitness when giant kelp is unavailable is unknown. We experimentally examined the effects of single and mixed species diets on consumption, growth and gonad weight in the purple sea urchin Strongylocentrotus purpuratus. Urchins were fed single species diets consisting of one of four common species of macroalgae (the kelps Macrocystis pyrifera and Pterygophora californica, and the red algae Chondracanthus corymbiferus and Rhodymenia californica (hereafter referred to by genus)) or a mixed diet containing all four species ad libitum over a 13-week period in a controlled laboratory setting. Urchins fed Chondracanthus, Macrocystis and a mixed diet showed the highest growth (in terms of test diameter, wet weight and jaw length) and gonad weight, while urchins fed Pterygophora and Rhodymenia showed the lowest. Urchins consumed their preferred food, Macrocystis, at the highest rate when offered a mixture, but consumed Chondracanthus or Macrocystis at similar rates when the two algae were offered alone. The differences in urchin feeding behavior and growth observed between these diet types suggest the relative availability of the algae tested here could affect urchin populations and their interactions with the algal assemblage. The fact that the performance of urchins fed Chondracanthus was similar or higher than those fed the preferred Macrocystis suggests that the availability of the former could could sustain growth and reproduction of purple sea urchins during times of low Macrocystis abundance as is common following

  19. Analysis of histone gene expression in adult tissues of the sea urchins Strongylocentrotus purpuratus and Lytechinus pictus: tissue-specific expression of sperm histone genes.

    PubMed Central

    Lieber, T; Weisser, K; Childs, G

    1986-01-01

    We analyzed the histone mRNA population found in several adult tissues of the sea urchin Strongylocentrotus purpuratus and in testis of Lytechinus pictus. Unique species of H1 and H2b mRNAs encoding the sperm-specific histone subtypes can be found exclusively in testis RNA. S. purpuratus contains two distinct testis-specific H1 transcripts, while L. pictus contains one such transcript. Each of these mRNAs is larger than either early or late embryonic H1 mRNAs. Other somatic adult tissues contain transcripts derived from members of the late embryonic H1 histone gene family. S. purpuratus contains one H2b transcript found exclusively in testis, while L. pictus contains two such H2b mRNAs. Similarly, in tissues other than testis, late H2b transcripts were found. While there is no sperm-specific H2a protein, a limited set of late histone H2a genes encoding primarily the H2a-beta subtype is expressed in testis. The majority of the H2a protein found in diploid adult tissues is also the H2a-beta subtype; however, the size of the H2a transcripts differs between testis and other tissues. We conclude that different members of the late H2a gene family are differentially expressed in embryos and adult tissues. We prepared and characterized cDNA clones encoding the sperm-specific H2b protein as well as the H2a-beta protein found in testis. Images PMID:3785204

  20. Identification and mapping of amplified fragment length polymorphism markers linked to shell color in bay scallop, Argopecten irradians irradians (Lamarck, 1819).

    PubMed

    Qin, Yanjie; Liu, Xiao; Zhang, Haibin; Zhang, Guofan; Guo, Ximing

    2007-01-01

    Amplified fragment length polymorphisms (AFLP) were used to study the inheritance of shell color in Argopecten irradians. Two scallops, one with orange and the other with white shells, were used as parents to produce four F(1) families by selfing and outcrossing. Eighty-eight progeny, 37 orange and 51 white, were randomly selected from one of the families for segregation and mapping analysis with AFLP and microsatellite markers. Twenty-five AFLP primer pairs were screened, yielding 1138 fragments, among which 148 (13.0%) were polymorphic in two parents and segregated in progeny. Six AFLP markers showed significant (P < 0.05) association with shell color. All six loci were mapped to one linkage group. One of the markers, F1f335, is completely linked to the gene for orange shell, which we designated as Orange1, without any recombination in the progeny we sampled. The marker was amplified in the orange parent and all orange progeny, but absent in the white parent and all the white progeny. The close linkage between F1f335 and Orange1 was validated using bulk segregation analysis in two natural populations, and all our data indicate that F1f335 is specific for the shell color gene, Orange1. The genomic mapping of a shell color gene in bay scallop improves our understanding of shell color inheritance and may contribute to the breeding of molluscs with desired shell colors. PMID:17160637

  1. Accumulation, Biotransformation, Histopathology and Paralysis in the Pacific Calico Scallop Argopecten ventricosus by the Paralyzing Toxins of the Dinoflagellate Gymnodinium catenatum

    PubMed Central

    Escobedo-Lozano, Amada Y.; Estrada, Norma; Ascencio, Felipe; Contreras, Gerardo; Alonso-Rodriguez, Rosalba

    2012-01-01

    The dinoflagellate Gymnodinium catenatum produces paralyzing shellfish poisons that are consumed and accumulated by bivalves. We performed short-term feeding experiments to examine ingestion, accumulation, biotransformation, histopathology, and paralysis in the juvenile Pacific calico scallop Argopecten ventricosus that consume this dinoflagellate. Depletion of algal cells was measured in closed systems. Histopathological preparations were microscopically analyzed. Paralysis was observed and the time of recovery recorded. Accumulation and possible biotransformation of toxins were measured by HPLC analysis. Feeding activity in treated scallops showed that scallops produced pseudofeces, ingestion rates decreased at 8 h; approximately 60% of the scallops were paralyzed and melanin production and hemocyte aggregation were observed in several tissues at 15 h. HPLC analysis showed that the only toxins present in the dinoflagellates and scallops were the N-sulfo-carbamoyl toxins (C1, C2); after hydrolysis, the carbamate toxins (epimers GTX2/3) were present. C1 and C2 toxins were most common in the mantle, followed by the digestive gland and stomach-complex, adductor muscle, kidney and rectum group, and finally, gills. Toxin profiles in scallop tissue were similar to the dinoflagellate; biotransformations were not present in the scallops in this short-term feeding experiment. PMID:22822356

  2. Seasonal variation of biochemical components in clam ( Saxidomus purpuratus Sowerby 1852) in relation to its reproductive cycle and the environmental condition of Sanggou Bay, China

    NASA Astrophysics Data System (ADS)

    Bi, Jinhong; Li, Qi; Zhang, Xinjun; Zhang, Zhixin; Tian, Jinling; Xu, Yushan; Liu, Wenguang

    2016-04-01

    Seasonal variation of biochemical components in clam ( Saxidomus purpuratus Sowerby 1852) was investigated from March 2012 to February 2013 in relation to environmental condition of Sanggou Bay and the reproductive cycle of clam. According to the histological analysis, the reproductive cycle of S. purpuratus includes two distinctive phases: a total spent and inactive stage from November to January, and a gametogenesis stage, including ripeness and spawning, during the rest of the year. Gametes were generated at a low temperature (2.1°C) in February. Spawning took place once a year from June to October. The massive spawning occurred in August when the highest water temperature and chlorophyll a level could be observed. The key biochemical components (glycogen, protein and lipid) in five tissues (gonad, foot, mantle, siphon and adductor muscle) were analyzed. The glycogen content was high before gametogenesis, and decreased significantly during the gonad development in the gonad, mantle and foot of both females and males, suggesting that glycogen was an important energy source for gonad development. The protein and lipid contents increased in the ovary during the gonad development, demonstrating that they are the major organic components of oocytes. The lipid and protein contents decreased in the testis, implying that they can provide energy and material for spermatogenesis. The results also showed that protein stored in the mantle and foot could support the reproduction after the glycogen was depleted.

  3. Natural variation and the capacity to adapt to ocean acidification in the keystone sea urchin Strongylocentrotus purpuratus.

    PubMed

    Kelly, Morgan W; Padilla-Gamiño, Jacqueline L; Hofmann, Gretchen E

    2013-08-01

    A rapidly growing body of literature documents the potential negative effects of CO2 -driven ocean acidification (OA) on marine organisms. However, nearly all this work has focused on the effects of future conditions on modern populations, neglecting the role of adaptation. Rapid evolution can alter demographic responses to environmental change, ultimately affecting the likelihood of population persistence, but the capacity for adaptation will differ among populations and species. Here, we measure the capacity of the ecologically important purple sea urchin Strongylocentrotus purpuratus to adapt to OA, using a breeding experiment to estimate additive genetic variance for larval size (an important component of fitness) under future high-pCO2 /low-pH conditions. Although larvae reared under future conditions were smaller than those reared under present-day conditions, we show that there is also abundant genetic variation for body size under elevated pCO2 , indicating that this trait can evolve. The observed heritability of size was 0.40 ± 0.32 (95% CI) under low pCO2 , and 0.50 ± 0.30 under high-pCO2 conditions. Accounting for the observed genetic variation in models of future larval size and demographic rates substantially alters projections of performance for this species in the future ocean. Importantly, our model shows that after incorporating the effects of adaptation, the OA-driven decrease in population growth rate is up to 50% smaller, than that predicted by the 'no-adaptation' scenario. Adults used in the experiment were collected from two sites on the coast of the Northeast Pacific that are characterized by different pH regimes, as measured by autonomous sensors. Comparing results between sites, we also found subtle differences in larval size under high-pCO2 rearing conditions, consistent with local adaptation to carbonate chemistry in the field. These results suggest that spatially varying selection may help to maintain genetic variation necessary

  4. The Polymorphism in the Promoter of HSP70 Gene Is Associated with Heat Tolerance of Two Congener Endemic Bay Scallops (Argopecten irradians irradians and A. i. concentricus)

    PubMed Central

    Yang, Chuanyan; Wang, Lingling; Wang, Jingjing; Jiang, Qiufen; Qiu, Limei; Zhang, Huan; Song, Linsheng

    2014-01-01

    Background The heat shock protein 70 (HSP70) is one kind of molecular chaperones, which plays a key role in protein metabolism under normal and stress conditions. Methodology In the present study, the mRNA expressions of HSP70 under normal physiological condition and after acute heat stress were investigated in gills of two bay scallop populations (Argopecten irradians irradians and A. i. concentricus). The heat resistant scallops A. i. concentricus showed significantly lower basal level and higher induction of HSP70 compared with that of the heat sensitive scallops A. i. irradians. The promoter sequence of HSP70 gene from bay scallop (AiHSP70) was cloned and the polymorphisms within this region were investigated to analyze their association with heat tolerance. Totally 11 single nucleotide polymorphisms (SNPs) were identified, and four of them (−967, −480, −408 and −83) were associated with heat tolerance after HWE analysis and association analysis. Based on the result of linkage disequilibrium analysis, the in vitro transcriptional activities of AiHSP70 promoters with different genotype were further determined, and the results showed that promoter from A. i. concentricus exhibited higher transcriptional activity than that from A. i. irradians (P<0.05). Conclusions The results provided insights into the molecular mechanisms underlying the thermal adaptation of different congener endemic bay scallops, which suggested that the increased heat tolerance of A. i. concentricus (compared with A. i. irradians) was associated with the higher expression of AiHSP70. Meanwhile, the −967 GG, −480 AA, −408 TT and −83 AG genotypes could be potential markers for scallop selection breeding with higher heat tolerance. PMID:25028964

  5. A detailed staging scheme for late larval development in Strongylocentrotus purpuratus focused on readily-visible juvenile structures within the rudiment

    PubMed Central

    2014-01-01

    Background The purple sea urchin, Strongylocentrotus purpuratus, has long been the focus of developmental and ecological studies, and its recently-sequenced genome has spawned a diversity of functional genomics approaches. S. purpuratus has an indirect developmental mode with a pluteus larva that transforms after 1–3 months in the plankton into a juvenile urchin. Compared to insects and frogs, mechanisms underlying the correspondingly dramatic metamorphosis in sea urchins remain poorly understood. In order to take advantage of modern techniques to further our understanding of juvenile morphogenesis, organ formation, metamorphosis and the evolution of the pentameral sea urchin body plan, it is critical to assess developmental progression and rate during the late larval phase. This requires a staging scheme that describes developmental landmarks that can quickly and consistently be used to identify the stage of individual living larvae, and can be tracked during the final two weeks of larval development, as the juvenile is forming. Results Notable structures that are easily observable in developing urchin larvae are the developing spines, test and tube feet within the juvenile rudiment that constitute much of the oral portion of the adult body plan. Here we present a detailed staging scheme of rudiment development in the purple urchin using soft structures of the rudiment and the primordia of these juvenile skeletal elements. We provide evidence that this scheme is robust and applicable across a range of temperature and feeding regimes. Conclusions Our proposed staging scheme provides both a useful method to study late larval development in the purple urchin, and a framework for developing similar staging schemes across echinoderms. Such efforts will have a high impact on evolutionary developmental studies and larval ecology, and facilitate research on this important deuterostome group. PMID:24886415

  6. A Consensus Microsatellite-Based Linkage Map for the Hermaphroditic Bay Scallop (Argopecten irradians) and Its Application in Size-Related QTL Analysis

    PubMed Central

    Li, Hongjun; Liu, Xiao; Zhang, Guofan

    2012-01-01

    Bay scallop (Argopecten irradians) is one of the most economically important aquaculture species in China. In this study, we constructed a consensus microsatellite-based genetic linkage map with a mapping panel containing two hybrid backcross-like families involving two subspecies of bay scallop, A. i. irradians and A. i. concentricus. One hundred sixty-one microsatellite and one phenotypic (shell color) markers were mapped to 16 linkage groups (LGs), which corresponds to the haploid chromosome number of bay scallop. The sex-specific map was 779.2 cM and 781.6 cM long in female and male, respectively, whereas the sex-averaged map spanned 849.3 cM. The average resolution of integrated map was 5.9 cM/locus and the estimated coverage was 81.3%. The proportion of distorted markers occurred more in the hybrid parents, suggesting that the segregation distortion was possibly resulted from heterospecific interaction between genomes of two subspecies of bay scallop. The overall female-to-male recombination rate was 1.13∶1 across all linked markers in common to both parents, and considerable differences in recombination also existed among different parents in both families. Four size-related traits, including shell length (SL), shell height (SH), shell width (SW) and total weight (TW) were measured for quantitative trait loci (QTL) analysis. Three significant and six suggestive QTL were detected on five LGs. Among the three significant QTL, two (qSW-10 and qTW-10, controlling SW and TW, respectively) were mapped on the same region near marker AiAD121 on LG10 and explained 20.5% and 27.7% of the phenotypic variance, while the third (qSH-7, controlling SH) was located on LG7 and accounted for 15.8% of the phenotypic variance. Six suggestive QTL were detected on four different LGs. The linkage map and size-related QTL obtained in this study may facilitate marker-assisted selection (MAS) in bay scallop. PMID:23077533

  7. The elemental composition of purple sea urchin (Strongylocentrotus purpuratus) calcite and potential effects of pCO2 during early life stages

    NASA Astrophysics Data System (ADS)

    LaVigne, M.; Hill, T. M.; Sanford, E.; Gaylord, B.; Russell, A. D.; Lenz, E. A.; Hosfelt, J. D.; Young, M. K.

    2013-06-01

    Ocean acidification will likely have negative impacts on invertebrates producing skeletons composed of calcium carbonate. Skeletal solubility is partly controlled by the incorporation of "foreign" ions (e.g. magnesium) into the crystal lattice of these skeletal structures, a process that is sensitive to a variety of biological and environmental factors. Here we explore effects of life stage, oceanographic region of origin, and changes in the partial pressure of carbon dioxide in seawater (pCO2) on trace elemental composition in the purple sea urchin (Strongylocentrotus purpuratus). We show that, similar to other urchin taxa, adult purple sea urchins have the ability to precipitate skeleton composed of a range of biominerals spanning low- to high-Mg calcites. Mg / Ca and Sr / Ca ratios were substantially lower in adult spines compared to adult tests. On the other hand, trace elemental composition was invariant among adults collected from four oceanographically distinct regions spanning a range of carbonate chemistry conditions (Oregon, Northern California, Central California, and Southern California). Skeletons of newly settled juvenile urchins that originated from adults from the four regions exhibited intermediate Mg / Ca and Sr / Ca between adult spine and test endmembers, indicating that skeleton precipitated during early life stages is more soluble than adult spines and less soluble than adult tests. Mean skeletal Mg / Ca or Sr / Ca of juvenile skeleton did not vary with source region when larvae were reared under present-day, global-average seawater carbonate conditions (400 μatm; pHT = 8.02 ± 0.03 1 SD; Ωcalcite = 3.3 ± 0.2 1 SD). However, when reared under elevated pCO2 (900 μatm; pHT = 7.73 ± 0.03; Ωcalcite = 1.8 ± 0.1), skeletal Sr / Ca in juveniles exhibited increased variance across the four regions. Although larvae from the northern populations (Oregon, Northern California, Central California) did not exhibit differences in Mg or Sr

  8. Precise cis-regulatory control of spatial and temporal expression of the alx-1 gene in the skeletogenic lineage of s. purpuratus.

    PubMed

    Damle, Sagar; Davidson, Eric H

    2011-09-15

    Deployment of the gene-regulatory network (GRN) responsible for skeletogenesis in the embryo of the sea urchin Strongylocentrotus purpuratus is restricted to the large micromere lineage by a double negative regulatory gate. The gate consists of a GRN subcircuit composed of the pmar1 and hesC genes, which encode repressors and are wired in tandem, plus a set of target regulatory genes under hesC control. The skeletogenic cell state is specified initially by micromere-specific expression of these regulatory genes, viz. alx1, ets1, tbrain and tel, plus the gene encoding the Notch ligand Delta. Here we use a recently developed high throughput methodology for experimental cis-regulatory analysis to elucidate the genomic regulatory system controlling alx1 expression in time and embryonic space. The results entirely confirm the double negative gate control system at the cis-regulatory level, including definition of the functional HesC target sites, and add the crucial new information that the drivers of alx1 expression are initially Ets1, and then Alx1 itself plus Ets1. Cis-regulatory analysis demonstrates that these inputs quantitatively account for the magnitude of alx1 expression. Furthermore, the Alx1 gene product not only performs an auto-regulatory role, promoting a fast rise in alx1 expression, but also, when at high levels, it behaves as an auto-repressor. A synthetic experiment indicates that this behavior is probably due to dimerization. In summary, the results we report provide the sequence level basis for control of alx1 spatial expression by the double negative gate GRN architecture, and explain the rising, then falling temporal expression profile of the alx1 gene in terms of its auto-regulatory genetic wiring. PMID:21723273

  9. Precise cis-regulatory control of spatial and temporal expression of the alx-1 gene in the skeletogenic lineage of s. purpuratus

    PubMed Central

    Damle, Sagar; Davidson, Eric H.

    2011-01-01

    Deployment of the gene regulatory network (GRN) responsible for skeletogenesis in the embryo of the sea urchin Strongylocentrotus purpuratus is restricted to the large micromere lineage by a double negative regulatory gate. The gate consists of a GRN subcircuit composed of the pmar1 and hesC genes, which encode repressors and are wired in tandem, plus a set of target regulatory genes under hesC control. The skeletogenic cell state is specified initially by micromere-specific expression of these regulatory genes, viz. alx1, ets1, tbrain and tel, plus the gene encoding the Notch ligand Delta. Here we use a recently developed high throughput methodology for experimental cis-regulatory analysis to elucidate the genomic regulatory system controlling alx1 expression in time and embryonic space. The results entirely confirm the double negative gate control system at the cis-regulatory level, including definition of the functional HesC target sites, and add the crucial new information that the drivers of alx1 expression are initially Ets1, and then Alx1 itself plus Ets1. Cis-regulatory analysis demonstrates that these inputs quantitatively account for the magnitude of alx1 expression. Furthermore, the Alx1 gene product not only performs an auto-regulatory role, promoting a fast rise in alx1 expression, but also, when at high levels, it behaves as an autorepressor. A synthetic experiment indicates that this behavior is probably due to dimerization. In summary, the results we report provide the sequence level basis for control of alx1 spatial expression by the double negative gate GRN architecture, and explain the rising, then falling temporal expression profile of the alx1 gene in terms of its auto-regulatory genetic wiring. PMID:21723273

  10. Effects of produced water on reproduction and early life stages of the purple sea urchin (Strongylocentrotus purpuratus): Field and laboratory tests

    SciTech Connect

    Krause, P.R.

    1993-01-01

    This dissertation focuses on the effects of produced water (an oil-production effluent) on reproduction in the purple sea urchin (Strongy-locentrotus purpuratus) using both field and laboratory experiments. The author investigated the effects of chronic exposure to produced water on the gametogenesis and gamete performance using an in-situ caging experiment. He found a significant negative relationship between gonad mass and cage distance for both sexes, indicating that urchins living closer to the outfall produced significantly larger gonads. He also found significant differences in the fertilizability of eggs between cages and this showed a positive relationship with distance from the outfall. These findings indicate that while urchins exposed to a produced water outfall produce large gonads, they suffer a marked decrease in gamete performance. In a subsequent study the author explored whether and how brief exposure to a range of concentrations of produced water affected gametes and early larval stages of the purple sea urchin. Specifically, he exposed separately and together, eggs, sperm, and zygotes to ascertain the relative sensitivities of these life stages to produced water at durations and concentrations realistic to each state. He also explored the nature of the biological responses, and the potential for delayed expression. I found that both apparent fertilization and embryonic developmental success showed decreased performance with increasing produced water concentrations. Produced water exposure effectively slowed embryological developmental rates, but did not affect embryo survivorship. The spatial and temporal variability in toxicity of receiving waters was addressed using a fertilization bioassay. Receiving waters were collected along a transect down-field from the discharge on three dates while the outfall was actively discharging, and on one date while the plant was not discharging.

  11. Introduction, spat-rearing and experimental culture of bay scallop, Argopecten irradians lamarck

    NASA Astrophysics Data System (ADS)

    Zhang, Fusui; He, Yichao; Liu, Xiangsheng; Ma, Jianghu; Li, Shuying; Qi, Lingxin

    1991-06-01

    Bay scallops were introduced from the east coast of the U.S. into China to shorten the economic turn-over from 2 to 1 year. Parent scallops were carried to Qingdao on Dec. 20, 1982 and stocked in indoor tanks at controlled temperature and fed with a mixture of Phaeodictylum tricornutum, Pyramimonas sp. and Chlorella sp. They spawned on Jan. 26 of the next year. The larvae were reared at a temperature of 18 21°C and fed with Isochrysis galbana, Pyramimonas sp. and Chorella sp. In 4 weeks’ growing, the spats averaged 827 μm. They attained a height of 6.9 mm on May 9. In the middle of May, the seed scallops were transferred to Luoyuan Bay in Fujian Province, and Jiaozhou Bay, as well as the area off Taiping Jiao Cape, Qingdao, Shandong Province for experimental culture in plastic netcages suspended on a single line raft. Bay scallops cultured in Luoyuan Bay grew to an averaged shell height of 10.4 mm. In Jiaozhou Bay and the Taiping Jiao Cape culture area, they grew to 50 mm in average shell height (marketable size) and 26 g in average weight by late September; and attained 59 mm in average shell height (R. 39 75 mm) and 46 g in average weight in late December. The ovary and testis could be distinguished by color in August. In early September, eggs and sperms were collected for our laboratory where the second generation of seed scallops was successfully reared to suitable size for growing outdoor and breeding. This species can be harvested within a year after fertilization of the eggs, so we consider it suitable for culture in the Yellow Sea and East China Sea. This is the first reported successful introduction of Atlantic mollusks to the China Seas.

  12. Effects of environmental oxygen deficiency on embryos and larvae of bay scallop, Argopecten irradians irradians

    NASA Astrophysics Data System (ADS)

    Chun-de, Wang; Fu-Sui, Zhang

    1995-12-01

    The experimental results showed that: 1) The embryonic development of bay scallop is inhibited at a dissolved oxygen range of 1.38-3.64×10-3 at 23°C, and completely blocked below the lower limit. 2) The tolerance of larvae to anoxia increased with larval sizes and was related to their oxygen debt. 3) The scallop larvae exhibited specific behavioral responses to oxygen deficiency, which finally led to velum disintegration and larval death. The possible relationship between environmental oxygen deficiency and the disease of disintegration of the larval velum is also discussed. In this study, considerable oxygen debt was found in bay scallop larvae, which was greater in small animals. Based on the works of previous authors, a new concept is proposed for the estimation of oxygen debt, namely, the compensatory rate of oxygen debt (CROD). This can be used in intra—or interspecific comparison of oxygen debt. The results can be helpful in the management of water quality and for the prevention of larval diseases encountered in scallop culture.

  13. ASSESSING THE EFFECTS OF HABITAT ALTERATION ON THE BAY SCALLOP, ARGOPECTEN IRRADIANS

    EPA Science Inventory

    The U.S. EPA's National Health and Environmental Effects Laboratory is evaluating approaches for protecting and restoring the ecological integrity of aquatic habitats, such as marine wetlands and coastal zones, from the impacts of multiple aquatic stressors. We are developing ha...

  14. Bioconversion of (13)C-labeled microalgal phytosterols to cholesterol by the Northern Bay scallop, Argopecten irradians irradians.

    PubMed

    Giner, José-Luis; Zhao, Hui; Dixon, Mark S; Wikfors, Gary H

    2016-02-01

    Bivalve mollusks lack de novo cholesterol biosynthesis capabilities and therefore rely upon dietary sources of sterols for rapid growth. Microalgae that constitute the main source of nutrition for suspension-feeding bivalves contain a diverse array of phytosterols, in most cases lacking cholesterol. Rapid growth of bivalves on microalgal diets with no cholesterol implies that some phytosterols can satisfy the dietary requirement for cholesterol through metabolic conversion to cholesterol, but such metabolic pathways have not been rigorously demonstrated. In the present study, stable isotope-labeled phytosterols were used to supplement a unialgal diet of Rhodomonas sp. and their biological transformation to cholesterol within scallop tissues was determined using (13)C-NMR spectroscopy. Scallops efficiently dealkylated ∆(5) C29 (24-ethyl) sterols to cholesterol, and the only C28 sterol that was dealkylated efficiently possessed the 24(28)-double bond. Non-metabolized dietary phytosterols accumulated in the soft tissues. Observed formation of ∆(5,7) sterols (provitamin D) from ∆(5) sterols may represent initiation of steroid hormone (possibly ecdysone) biosynthesis. These findings provide a key component necessary for formulation of nutritionally complete microalgal diets for hatchery production of seed for molluscan aquaculture. PMID:26577022

  15. Sublethal mechanisms of Pb and Zn toxicity to the purple sea urchin (Strongylocentrotus purpuratus) during early development.

    PubMed

    Tellis, Margaret S; Lauer, Mariana M; Nadella, Sunita; Bianchini, Adalto; Wood, Chris M

    2014-01-01

    In order to understand sublethal mechanisms of lead (Pb) and zinc (Zn) toxicity, developing sea urchins were exposed continuously from 3h post-fertilization (eggs) to 96 h (pluteus larvae) to 55 (±2.4) μgPb/L or 117 (±11)μgZn/L, representing ~ 70% of the EC50 for normal 72 h development. Growth, unidirectional Ca uptake rates, whole body ion concentrations (Na, K, Ca, Mg), Ca(2+) ATPase activity, and metal bioaccumulation were monitored every 12h over this period. Pb exhibited marked bioaccumulation whereas Zn was well-regulated, and both metals had little effect on growth, measured as larval dry weight, or on Na, K, or Mg concentrations. Unidirectional Ca uptake rates (measured by (45)Ca incorporation) were severely inhibited by both metals, resulting in lower levels of whole body Ca accumulation. The greatest disruption occurred at gastrulation. Ca(2+) ATPase activity was also significantly inhibited by Zn but not by Pb. Interestingly, embryos exposed to Pb showed some capacity for recovery, as Ca(2+)ATPase activities increased, Ca uptake rates returned to normal intermittently, and whole body Ca levels were restored to control values by 72-96 h of development. This did not occur with Zn exposure. Both Pb and Zn rendered their toxic effects through disruption of Ca homeostasis, though likely through different proximate mechanisms. We recommend studying the toxicity of these contaminants periodically throughout development as an effective way to detect sublethal effects, which may not be displayed at the traditional toxicity test endpoint of 72 h. PMID:24326189

  16. Induction of DNA-protein cross-links in developing embryos of the purple sea urchin, Strongylocentrotus purpuratus

    SciTech Connect

    Garman, G.D.; Cherr, G.N.; Anderson, S.L.

    1994-12-31

    Exposure to environmental agents during embryonic development may result in DNA-protein cross-linking (DPC), as has been demonstrated for mammalian cell lines. In the latter, formation of DPC`s upon exposure to a wide variety of agents, including some metals, has been observed. To determine whether DPCs could be detected in the sea urchin embryo during development, the authors adapted a mammalian cell assay utilizing potassium-SDS precipitation and a DNA fluorochrome to quantify relative amounts of free and protein-bound DNA. Sea urchin embryos exposed to a known DPC agent, nickel, through gastrulation exhibited a dose-dependent increase in DPCs, as well as an increase in developmental abnormalities. Morphological studies demonstrated that stage-specific exposure to Ni prior to gastrulation resulted in similar levels of abnormal pluteus larval development as compared to embryos exposed through gastrulation. Sea urchin embryos exhibit temporal differences in DNA transcription and gene expression during development, and these could be affected by modifications in DNA-protein interactions. Therefore, the authors are investigating the hypothesis that the similarities in morphological responses observed may relate to susceptibility of a critical stage of development.

  17. Residual Circulation in a Chilean Equatorward Facing Embayment: Tongoy Bay

    NASA Astrophysics Data System (ADS)

    Moraga-Opazo, J.; Valle-Levinson, A.

    2006-12-01

    Underway current velocity profiles were measured along a pair of transects during a period of 48 hours in an equatorward facing bay with the objective of characterizing the impact of circulation in bay aquaculture activities. The bay is Tongoy, located in north-central Chile (30° 15´S, 71° 35´W) and oriented northward. The bay entrance is oriented in the east-west direction and its width is 12 km while its maximum depth is 100 m. The bay is near a permanent upwelling focus that is likely to supply nutrients. The eastern portion of the bay is used for aquaculture (suspended system) of the scallop Argopecten purpuratus and is the most important aquaculture area in northern Chile. The velocity measurements were obtained during austral autumn and spring, in April and December 2005. Mean flows showed inflow over the eastern side of the entrance and outflow on the western side, following the bay's morphology. The mean flows were almost unidirectional in the autumn but changed more dramatically with depth in the spring. Results of a simple analytical model that compares the relative influence of frictional and Earth's rotational effects suggest that these mean circulations were close to being in geostrophic balance. This meant that frictional influences were negligible, corresponding to a low Ekman number regime. The mean circulation suggested that larvae should be concentrated near the coastline, in depths < 20 m, with greatest abundances to the southwest of the bay.

  18. Permanent genetic resources added to Molecular Ecology Resources Database 1 August 2011-30 September 2011.

    PubMed

    A'Hara, S W; Amouroux, P; Argo, Emily E; Avand-Faghih, A; Barat, Ashoktaru; Barbieri, Luiz; Bert, Theresa M; Blatrix, R; Blin, Aurélie; Bouktila, D; Broome, A; Burban, C; Capdevielle-Dulac, C; Casse, N; Chandra, Suresh; Cho, Kyung Jin; Cottrell, J E; Crawford, Charles R; Davis, Michelle C; Delatte, H; Desneux, Nicolas; Djieto-Lordon, C; Dubois, M P; El-Mergawy, R A A M; Gallardo-Escárate, C; Garcia, M; Gardiner, Mary M; Guillemaud, Thomas; Haye, P A; Hellemans, B; Hinrichsen, P; Jeon, Ji Hyun; Kerdelhué, C; Kharrat, I; Kim, Ki Hwan; Kim, Yong Yul; Kwan, Ye-Seul; Labbe, Ellen M; LaHood, Eric; Lee, Kyung Mi; Lee, Wan-Ok; Lee, Yat-Hung; Legoff, Isabelle; Li, H; Lin, Chung-Ping; Liu, S S; Liu, Y G; Long, D; Maes, G E; Magnoux, E; Mahanta, Prabin Chandra; Makni, H; Makni, M; Malausa, Thibaut; Matura, Rakesh; McKey, D; McMillen-Jackson, Anne L; Méndez, M A; Mezghani-Khemakhem, M; Michel, Andy P; Paul, Moran; Muriel-Cunha, Janice; Nibouche, S; Normand, F; Palkovacs, Eric P; Pande, Veena; Parmentier, K; Peccoud, J; Piatscheck, F; Puchulutegui, Cecilia; Ramos, R; Ravest, G; Richner, Heinz; Robbens, J; Rochat, D; Rousselet, J; Saladin, Verena; Sauve, M; Schlei, Ora; Schultz, Thomas F; Scobie, A R; Segovia, N I; Seyoum, Seifu; Silvain, J-F; Tabone, Elisabeth; Van Houdt, J K J; Vandamme, S G; Volckaert, F A M; Wenburg, John; Willis, Theodore V; Won, Yong-Jin; Ye, N H; Zhang, W; Zhang, Y X

    2012-01-01

    This article documents the addition of 299 microsatellite marker loci and nine pairs of single-nucleotide polymorphism (SNP) EPIC primers to the Molecular Ecology Resources (MER) Database. Loci were developed for the following species: Alosa pseudoharengus, Alosa aestivalis, Aphis spiraecola, Argopecten purpuratus, Coreoleuciscus splendidus, Garra gotyla, Hippodamia convergens, Linnaea borealis, Menippe mercenaria, Menippe adina, Parus major, Pinus densiflora, Portunus trituberculatus, Procontarinia mangiferae, Rhynchophorus ferrugineus, Schizothorax richardsonii, Scophthalmus rhombus, Tetraponera aethiops, Thaumetopoea pityocampa, Tuta absoluta and Ugni molinae. These loci were cross-tested on the following species: Barilius bendelisis, Chiromantes haematocheir, Eriocheir sinensis, Eucalyptus camaldulensis, Eucalyptus cladocalix, Eucalyptus globulus, Garra litaninsis vishwanath, Garra para lissorhynchus, Guindilla trinervis, Hemigrapsus sanguineus, Luma chequen. Guayaba, Myrceugenia colchagüensis, Myrceugenia correifolia, Myrceugenia exsucca, Parasesarma plicatum, Parus major, Portunus pelagicus, Psidium guayaba, Schizothorax richardsonii, Scophthalmus maximus, Tetraponera latifrons, Thaumetopoea bonjeani, Thaumetopoea ispartensis, Thaumetopoea libanotica, Thaumetopoea pinivora, Thaumetopoea pityocampa ena clade, Thaumetopoea solitaria, Thaumetopoea wilkinsoni and Tor putitora. This article also documents the addition of nine EPIC primer pairs for Euphaea decorata, Euphaea formosa, Euphaea ornata and Euphaea yayeyamana. PMID:22136175

  19. The occurrence of domoic acid linked to a toxic diatom bloom in a new potential vector: the tunicate Pyura chilensis (piure).

    PubMed

    López-Rivera, Américo; Pinto, Maricela; Insinilla, Andrea; Suárez Isla, Benjamín; Uribe, Eduardo; Alvarez, Gonzalo; Lehane, Mary; Furey, Ambrose; James, Kevin J

    2009-11-01

    The tunicate Pyura chilensis (Molina, 1782); Phylum Chordata; Subphylum Urochordata; Class Ascidiacea, common local name "piure" or sea squirt; a filter-feeder (plankton and suspended particles) sessile species; may play an important role in monitoring domoic acid (DA) the principal toxic component of Amnesic Shellfish Poisoning (ASP). Significant DA concentrations have been determined in tunicate samples, collected during a recent ASP outbreak in Bahía Inglesa, an important scallop (Argopecten purpuratus) farming area. Several infaunal species were tested for the presence of DA, in addition to the usual scallop monitoring programme. DA was found at sub-toxic levels in filtering bivalves such as mussels (Mytilus chilensis), large mussels (Aulacomya ater) and clams (Protothaca thaca) (6.4, 5.4 and 4.7 microg DA/g tissue respectively). Of particular interest was the observation of significant accumulations of toxic Pseudo-nitzschia sp. diatoms in the internal siphon and atrium spaces of the tunicate. Toxin distribution within major tunicate organs was heterogeneous with 8.7-15.5 microg DA/g in edible tissues, 14.9-17.9 microg DA/g in the fecal material and 13.6-32.7 microg DA/g in the gut content. DA was determined by HPLC-UV and confirmed by diode-array detection and LC-MS/MS analysis. This is the first report of the presence of DA in a tunicate that is regularly consumed by coastal populations. These results confirm the need to include these organisms in sanitation programs for marine toxins. PMID:19505493

  20. Trophic and environmental drivers of the Sechura Bay Ecosystem (Peru) over an ENSO cycle

    NASA Astrophysics Data System (ADS)

    Taylor, Marc H.; Wolff, Matthias; Vadas, Flora; Yamashiro, Carmen

    2008-03-01

    Interannual environmental variability in Peru is dominated by the El Niño Southern Oscillation (ENSO). The most dramatic changes are associated with the warm El Niño (EN) phase (opposite the cold La Niña phase), which disrupts the normal coastal upwelling and affects the dynamics of many coastal marine and terrestrial resources. This study presents a trophic model for Sechura Bay, located at the northern extension of the Peruvian upwelling system, where ENSO-induced environmental variability is most extreme. Using an initial steady-state model for the year 1996, we explore the dynamics of the ecosystem through the year 2003 (including the strong EN of 1997/98 and the weaker EN of 2002/03). Based on support from literature, we force biomass of several non-trophically-mediated ‘drivers’ (e.g. Scallops, Benthic detritivores, Octopus, and Littoral fish) to observe whether the fit between historical and simulated changes (by the trophic model) is improved. The results indicate that the Sechura Bay Ecosystem is a relatively inefficient system from a community energetics point of view, likely due to the periodic perturbations of ENSO. A combination of high system productivity and low trophic level target species of invertebrates (i.e. scallops) and fish (i.e. anchoveta) results in high catches and an efficient fishery. The importance of environmental drivers is suggested, given the relatively small improvements in the fit of the simulation with the addition of trophic drivers on remaining functional groups’ dynamics. An additional multivariate regression model is presented for the scallop Argopecten purpuratus, which demonstrates a significant correlation between both spawning stock size and riverine discharge-mediated mortality on catch levels. These results are discussed in the context of the appropriateness of trophodynamic modeling in relatively open systems, and how management strategies may be focused given the highly environmentally influenced marine

  1. The 185/333 gene family is a rapidly diversifying host-defense gene cluster in the purple sea urchin Strongylocentrotus purpuratus.

    PubMed

    Buckley, K M; Munshaw, S; Kepler, T B; Smith, L C

    2008-06-13

    The genome of the purple sea urchin contains numerous large gene families with putative immunological functions. One gene family, known as 185/333, is characterized by extraordinary molecular diversity resulting from single nucleotide polymorphisms and the presence or the absence of 27 large blocks of sequences known as elements. The mosaic composition of elements, known as element patterns, that is present within the members of this gene family is encoded entirely in the second of two exons. Many of the elements correspond to one of six types of repeats that are present throughout the genes. The sequence diversity and variation in element patterns led us to investigate the evolution of the 185/333 gene family. The work presented here suggests that the element patterns are the result of both recombination and duplication and/or deletion of intragenic repeats. Each element is composed of a limited number of similar but distinct sequences, and their distribution among the 185/333 genes suggests frequent recombination within this gene family. Phylogenetic analyses of five 185/333 elements and two regions of the intron were performed using two tests: incongruence length difference and incongruence permutation. Results indicated that each pair of sequence segments was incongruent, suggesting that recombination occurs frequently along the length of the genes, including both the intron and the second exon, and that recombination is not restricted to intact elements. Paradoxically, the high level of similarity among the elements indicated that the 185/333 genes appear to be the result of a recent diversification. These results add to the growing body of evidence suggesting that invertebrate immune systems are not simple and static, but are dynamic and highly complex, and may employ group-specific mechanisms for diversification. PMID:18482736

  2. Copper transfer and influence on a marine food chain

    SciTech Connect

    Edding, M.; Tala, F.

    1996-12-31

    Copper is an essential element, required for normal growth by all plants and animals; and a regular constituent in the environment (Lewis and Cave 1982; Lewis 1994). This heavy metal is an essential micronutrient that at higher concentrations can be deleterious to algae and other aquatic biota (Chang and Sibley 1993). Copper toxicity to algae depends upon the individual species, their physiological and environmental conditions, and the chemical forms of metal in the medium (Sunda and Gullard 1976). When copper is accumulated by phytoplankton it can be transferred and may produce toxic effects on zooplankton (Wikfors and Ukeles 1982). Different species of microalgae present different capacities of resistance to copper. Cyanophyceae pre-cultured in a Cu-enriched medium (635 {mu}gCu{center_dot}L{sup {minus}1}) showed an EC{sub 50} that could reach 318 {mu}gCu{center_dot}L{sup {minus}1} for Plectonema radiosum and 339 {mu}gCu{center_dot}L{sup {minus}1} in Phormidium sp. (Takamura et al. 1990). Scenedesmus, Selenastrum and Chlorella were reported able to accumulate copper and other metal ions with an efficiency of 67-98% (Brady et al. 1994). Also, Dunaliella resisted concentrations form 0.38 mgCu{center_dot}L{sup {minus}1} (D. minuta) up to 50.8 mgCu{center_dot}L{sup {minus}1} (D. acidophila), depending on the pH of the medium (Grimmler et al. 1991). One the microalgae are copper-enriched, the copper that is part of the cell can be transferred to the surrounding water and to its predator producing uncertain effects. This study observed the effect of copper on the growth of Dunaliella tertiolecta and Isochrysis galbana that are currently used as food for hatchery-grown scallop larvae (Argopecten purpuratus). We observed the path of copper form the water column into the microalgal cell and the effect of copper-enriched food on the scallop larvae. 16 refs., 3 figs., 2 tabs.

  3. Distinctive expression patterns of 185/333 genes in the purple sea urchin, Strongylocentrotus purpuratus: an unexpectedly diverse family of transcripts in response to LPS, β-1,3-glucan, and dsRNA

    PubMed Central

    Terwilliger, David P; Buckley, Katherine M; Brockton, Virginia; Ritter, Nicole J; Smith, L Courtney

    2007-01-01

    Background A diverse set of transcripts called 185/333 is strongly expressed in sea urchins responding to immune challenge. Optimal alignments of full-length 185/333 cDNAs requires the insertion of large gaps that define 25 blocks of sequence called elements. The presence or absence of individual elements also defines a specific element pattern for each message. Individual sea urchins were challenged with pathogen associated molecular patterns (PAMPs) (lipopolysaccharide, β-1,3-glucan, or double stranded RNA), and changes in the 185/333 message repertoire were followed over time. Results Each animal expressed a diverse set of 185/333 messages prior to challenge and a 0.96 kb message was the predominant size after challenge. Sequence analysis of the cloned messages indicated that the major element pattern expressed in immunoquiescent sea urchins was either C1 or E2.1. In contrast, most animals responding to lipopolysaccharide, β-1,3-glucan or injury, predominantly expressed messages of the E2 pattern. In addition to the major patterns, extensive element pattern diversity was observed among the different animals before and after challenge. Nucleotide sequence diversity of the transcripts increased in response to β-1,3-glucan, double stranded RNA and injury, whereas diversity decreased in response to LPS. Conclusion These results illustrate that sea urchins appear to be able to differentiate among different PAMPs by inducing the transcription of different sets of 185/333 genes. Furthermore, animals may share a suite of 185/333 genes that are expressed in response to common pathogens, while also maintaining a large number of unique genes within the population. PMID:17331248

  4. SpTie1/2 is expressed in coelomocytes, axial organ and embryos of the sea urchin Strongylocentrotus purpuratus, and is an orthologue of vertebrate Tie1 and Tie2.

    PubMed

    Stevens, Margaret E; Dhillon, Jatinder; Miller, Chase A; Messier-Solek, Cynthia; Majeske, Audrey J; Zuelke, Dustin; Rast, Jonathan P; Smith, L Courtney

    2010-08-01

    A full length cDNA sequence expressed in coelomocytes shows significant sequence match to vertebrate Tie1 and Tie2/TEK. Vertebrate Tie2/TEK is the receptor for the angiopoietins and plays an important role in angiogenesis and hematopoiesis, whereas Tie1 regulates the activity of Tie2. The deduced sequence of the SpTie1/2 protein has a similar order and organization of domains to the homologous vertebrate proteins including a highly conserved receptor tyrosine kinase domain in the cytoplasmic tail. The N terminus of the ectodomain has one immunoglobulin (Ig)-Tie2_1 domain, followed by an Ig domain, four epidermal growth factor domains, a second Ig domain, and three fibronectin type III domains. The SpTie1/2 gene is expressed in coelomocytes and the axial organ, whereas other organs do not show significant expression. The timing of embryonic expression corresponds with the differentiation of blastocoelar cells, the embryonic and larval immune cells. Searches of the sea urchin genome show several gene models encoding putative ligands and signaling proteins that might interact with SpTie1/2. We speculate that SpTie1/2 may be involved in the proliferation of sea urchin immune cells in both adults and embryos. PMID:20363251

  5. Restoration Lessons Learned from Bay Scallop Habitat Models

    EPA Science Inventory

    Habitat quality and quantity are important factors to consider when restoring bay scallop (Argopecten irradians) populations; however, data linking habitat attributes to bay scallop populations are lacking. This information is essential to guide restoration efforts to reverse sc...

  6. RECENT BIOGENIC PHOSPHORITE: CONCRETIONS IN MOLLUSK KIDNEYS

    EPA Science Inventory

    Phosphorite concretions have been detected in the kidneys of two widespread species of mollusks. Mercenaria mercenaria and Argopecten irradians, which have relatively high population densities. These concretions are the first documentation of the direct biogenic formation of phos...

  7. CADMIUM TOXICITY TO THREE SPECIES OF ESTUARINE INVERTEBRATES

    EPA Science Inventory

    Three species of estuarine invertebrates, Palaemonetes pugio (grass shrimp), Pagurus longicarpus (hermit crab) and Argopecten irradians (bay scallop), were exposed to Cd in flowing seawater at concentrations of 0.06, 0.12, 0.25, 0.5 and 1.0 mg/litre. Incipient LC50 values of 0.53...

  8. Using a System Level Approach to Bay Scallop Enhancement and Management

    EPA Science Inventory

    Within the last twenty years total harvests of bay scallops (Argopecten irradians) have dwindled in the southern New England Region. Management of scallop populations has been underway on Martha’s Vineyard Island to provide the public with a fishery, and have included a com...

  9. STATUS AND APPLICATIONS OF ECHINOID (PHYLUM ECHINODERMATA) TOXICITY TEST METHODS

    EPA Science Inventory

    The use of echinoderms for toxicity testing has focused primarily on sea urchins and sand dollars (Strongylocentrolus purpuratus, Arbacia punctulata, Lytechinus pictus, and Dendraster excentricus, for example). he status and relative sensitivity of various test methods are descri...

  10. Recent biogenic phosphorite: Concretions in mollusk kidneys

    USGS Publications Warehouse

    Doyle, L.J.; Blake, N.J.; Woo, C.C.; Yevich, P.

    1978-01-01

    Phosphorite concretions have been detected in the kidneys of two widespread species ofmollusks, Mercenaria mercenaria and Argopecten irradians, which have relatively high population densities. These concretions are thefirst documentation of the direct biogenic formation of phosphorite grains. The concretions are principally amorphous calcium phosphate, which upon being heated yields an x-ray diffraction pattern which is essentially that of chlorapatite. These concretions appear to be a normal formation of the excretory process of mollusks under reproductive, environmental, or pollutant-induced stress. Biogenic production of phosphorite concretions over long periods of time and diagenetic change from amorphous to crystalline structure, coupled with secondary enrichment, may account for the formation of some marine phosphorite desposits which are not easily explained by the chemical precipitation- replacement hypothesis. Copyright ?? 1978 AAAS.

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

    PubMed

    Kolmann, Matthew A; Huber, Daniel R

    2009-01-01

    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, 19-59cm 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 (8-163N) and posterior (15-382N) 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 430N. 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. PMID:19428230

  12. Toxicity of lead and zinc to developing mussel and sea urchin embryos: critical tissue residues and effects of dissolved organic matter and salinity.

    PubMed

    Nadella, Sunita R; Tellis, Margaret; Diamond, Rachael; Smith, Scott; Bianchini, Adalto; Wood, Chris M

    2013-08-01

    Lead (Pb) EC50 values in the very sensitive early development phases (48-72h post-fertilization) of the mussels Mytilus galloprovincialis and Mytilus trossolus and sea urchin Strongylocentrotus purpuratus in 100% sea water were: M. trossolus - 45 (95% C.I.=22-72) μgL(-1); M. galloprovincialis - 63 (36-94) μgL(-1); S. purpuratus - 74 (50-101) μgL(-1). Salinity thresholds for normal development varied: M. trossolus>21ppt; M. galloprovincialis>28ppt; S. purpuratus≥30ppt. Addition of two spectroscopically distinct dissolved organic matters (DOM) from fresh water (Nordic Reservoir) and sea water (Inshore) moderately decreased the toxicity of Pb to both mussels, but not in a concentration-dependent fashion, with only an approximate doubling of EC50 over the range of 1.4-11.2mgCL(-1). Independent Pb binding capacity determinations for DOC explained the lack of a relationship between DOM concentration and toxicity. Salinity had no effect on Pb toxicity down to 21ppt in M. trossolus, and low salinity (21ppt) did not enhance the protective effect of DOC. Both DOMs increased the toxicity of Pb in developing sea urchin embryos, in contrast to mussels. Relative to Pb, the organisms were 6-9 fold less sensitive to Zn on a molar basis in 100% seawater with the following Zn EC50s: M. trossolus - 135 (103-170) μgL(-1); M. galloprovincialis - 172 (126-227) μgL(-1), S. purpuratus - 151 (129-177) μgL(-1). Nordic Reservoir and Inshore DOM (2-12mgCL(-1)) had no significant effect on Zn toxicity to mussels, in accord with voltammetry data showing an absence of any strong ligand binding for Zn by DOMs. As with Pb, DOMs increased Zn toxicity to urchin larvae. Critical Tissue Residues (CTR) based on whole body concentrations of Pb and Zn were determined for M. galloprovincialis at 48h and S. purpuratus at 72h. The median lethal CTR values (LA50s), useful parameters for development of saltwater Biotic Ligand Models (BLMs), were approximately 4-fold higher on a molar basis for Zn than

  13. [Genome sequencing in the sea urchin embryo: what is new concerning the cell cycle?].

    PubMed

    Genevière, Anne-Marie; Aze, Antoine; Even, Yasmine

    2007-01-01

    Sea urchin is a classical research model system in developmental biology; moreover, the external fertilization and growth of embryos, their rapid division cycle, their transparency and the accessibility of these embryos to molecular visualization methods, made them good specimens to analyze the regulatory mechanisms of cell division. These features as well as the phylogenetic position of sea urchin, close to vertebrates but in an outgroup within the deuterostomes, led scientists working on this model to sequence the genome of the species S. purpuratus. The genome contains a full repertoire of cell cycle control genes. A comparison of this toolkit with those from vertebrates, nematodes, drosophila, as well as tunicates, provides new insight into the evolution of cell cycle control. While some gene subtypes have undergone lineage-specific expansions in vertebrates (i.e. cyclins, mitotic kinases,...), others seem to be lost in vertebrates, for instance the novel cyclin B identified in S. purpuratus. On the other hand, some genes which were previously thought to be vertebrate innovations, are also found in sea urchins (i.e. MCM9). To note is also the absence of cell cycle inhibitors of the INK type, which are apparently confined to vertebrates. The uncovered genomic repertoire of cell-cycle regulators will thus provide molecular tools that should further enhance future research on cell cycle control and developmental regulation in this model. PMID:17762822

  14. Mytilid mussels: global habitat engineers in coastal sediments

    NASA Astrophysics Data System (ADS)

    Buschbaum, Christian; Dittmann, Sabine; Hong, Jae-Sang; Hwang, In-Seo; Strasser, Matthias; Thiel, Martin; Valdivia, Nelson; Yoon, San-Pil; Reise, Karsten

    2009-03-01

    Dense beds of mussels of the family Mytilidae occur worldwide on soft-bottoms in cold and warm temperate coastal waters and have usually been considered hot spots of biodiversity. We examined intertidal mussel beds at four distant locations around the globe with the same sampling method, to find out whether this “hot spot” designation holds universally. We studied species assemblages within the matrices of byssally interconnected mussels engineered by Mytilus edulis in the North Sea, by mixed Perumytilus purpuratus and Mytilus chilensis at the southern Chilean coast, by Musculista senhousia in the Yellow Sea and by Xenostrobus inconstans at the coast of southern Australia. In all cases, species assemblages inside mussel beds were significantly different from those outside with many species being restricted to one habitat type. However, species richness and diversity were not generally higher in mussel beds than in ambient sediments without mussels. In the North Sea ( M. edulis) and at the Chilean coast ( P. purpuratus, M. chilensis), mussel beds have markedly higher species numbers and diversities than surrounding sediments, but this was not the case for mussel beds in Australia ( X. inconstans) and the Yellow Sea ( M. senhousia) where numbers of associated species were only slightly higher and somewhat lower than in adjacent sediments, respectively. In conclusion, although soft bottom mytilid mussels generally enhance habitat heterogeneity and species diversity at the ecosystem level, mussel beds themselves are not universal centres of biodiversity, but the effects on associated species are site specific.

  15. A comparison of the copper sensitivity of six invertebrate species in ambient salt water of varying dissolved organic matter concentrations.

    PubMed

    Arnold, W Ray; Cotsifas, Jeffrey S; Ogle, R Scott; Depalma, Sarah G S; Smith, D Scott

    2010-02-01

    The copper sensitivity of four saltwater invertebrates (the mussel Mytilus galloprovincialis, the oyster Crassostrea virginica, the sand dollar Dendraster excentricus, and the sea urchin Strongylocentrotus purpuratus) was determined experimentally using chronic-estimator embryo-larval test procedures. The effect of sample dissolved organic matter (DOM) content on Cu bioavailability was determined for these species using commonly prescribed test procedures. Comparisons were made among these test results and test results reported previously for two other invertebrate species: the mussel Mytilus edulis and the copepod Eurytemora affinis. All six species exhibited a direct and significant relationship between the sample dissolved organic carbon (DOC; a surrogate measure of DOM) and either the dissolved Cu median lethal concentration (LC50) values or median effect concentration (EC50) values. This relationship is significant even when the DOM has different quality as evidenced by molecular fluorescence spectroscopy. Once normalized for the effects of DOM, the Cu sensitivity of these species from least to most sensitive were E. affinis < D. excitricus < C. virginica approximately S. purpuratus approximately M. edulis approximately M. galloprovincialis. This ranking of species sensitivity differs from the saltwater species sensitivity distribution proposed in 2003 by the U.S. Environmental Protection Agency. These results support the need to account for factors that modify Cu bioavailability in future saltwater Cu criteria development efforts. More specifically, Cu saltwater species sensitivity distribution data will need to be normalized by factors affecting Cu bioavailability to assure that accurate and protective criteria are subsequently developed for saltwater species and their uses. PMID:20821449

  16. Sensitivity and precision of whole effluent toxicity (WET) tests

    SciTech Connect

    Denton, D.; Chapman, G.; Fulk, F.

    1995-12-31

    The US Environmental Protection Agency`s test method manuals recommend reference toxicant test be performed to determine test sensitivity and precision within a test and among tests over time. The levels of intra and interlaboratory precision with two reference toxicants (zinc and copper) were examined with six marine test species (Macrocystis pyrifera, Haliotis rufescens, Strongylocentrotus purpuratus, Dendraster excentricus, Mytilus spp., and Menidia beryllina) and with two freshwater test species (Ceriodaphnia dubia and Pimephales promelas). Data from the EPA`s Reference Toxicant Database were used in the analysis. Data was used if the tests met the specified test acceptability criteria. The test sensitivity was examined by calculation of the minimum significant difference (e.g. MSD) and will be discussed. Results were compared to the national final chronic value (FCV) for copper and zinc. Greater than 99% of the EC25 values were above the FCV for copper with Strongylocentrotus purpuratus, Dendraster excentricus and Macrocystis pyrifera. However, greater than 99% of the EC25 values were below the FCV for zinc with Haliotis rufescens.

  17. Annual reversible plasticity of feeding structures: cyclical changes of jaw allometry in a sea urchin

    PubMed Central

    Ebert, Thomas A.; Hernández, José Carlos; Clemente, Sabrina

    2014-01-01

    A wide variety of organisms show morphologically plastic responses to environmental stressors but in general these changes are not reversible. Though less common, reversible morphological structures are shown by a range of species in response to changes in predators, competitors or food. Theoretical analysis indicates that reversible plasticity increases fitness if organisms are long-lived relative to the frequency of changes in the stressor and morphological changes are rapid. Many sea urchin species show differences in the sizes of jaws (demi-pyramids) of the feeding apparatus, Aristotle's lantern, relative to overall body size, and these differences have been correlated with available food. The question addressed here is whether reversible changes of relative jaw size occur in the field as available food changes with season. Monthly samples of the North American Pacific coast sea urchin Strongylocentrotus purpuratus were collected from Gregory Point on the Oregon (USA) coast and showed an annual cycle of relative jaw size together with a linear trend from 2007 to 2009. Strongylocentrotus purpuratus is a long-lived species and under field conditions individuals experience multiple episodes of changes in food resources both seasonally and from year to year. Their rapid and reversible jaw plasticity fits well with theoretical expectations. PMID:24500161

  18. The life cycle of Prosorhynchoides carvajali (Trematoda: Bucephalidae) involving species of bivalve and fish hosts in the intertidal zone of central Chile.

    PubMed

    Muñoz, G; Valdivia, I; López, Z

    2015-09-01

    We describe the life cycle of the bucephalid Prosorhynchoides carvajali from the intertidal rocky zone of central Chile. To elucidate the life cycle of this digenean, two mytilid bivalves, Semimytilus algosus and Perumytilus purpuratus, and ten intertidal fish species belonging to the families Blenniidae, Tripterygiidae, Labrisomidae, Kyphosidae and Gobiesocidae were analysed for natural infections. In addition, experimental infections of fish were undertaken and molecular analyses were performed of several developmental stages of the digeneans in various host species. Experimental infections of fish were made from infected mytilids to determine which fish species were suitable for the metacercarial stage of Prosorhynchoides. We also determined the abundance and prevalence of metacercariae in natural infections in fish and found that they were lower than in the experimental infections. A molecular analysis showed that sporocysts from S. algosus were identical to metacercariae from five fish species and P. carvajali adults. Sporocysts isolated from P. purpuratus were similar to metacercaria found in one fish species only (G. laevifrons) but were different from P. carvajali, with 1.9-2.0% genetic divergence. Therefore, the complete life cycle of P. carvajali consists of the mytilid species S. algosus as the first intermediate host, at least five intertidal fish species as second intermediate hosts (Scartichthys viridis, Auchenionchus microcirrhis, Hypsoblennius sordidus, Helcogrammoides chilensis and Gobiesox marmoratus), two carnivorous fish as definitive hosts (Auchenionchus microcirrhis and A. variolosus) and one occasional definitive host (Syciases sanguineus). This is the second description of a life cycle of a marine digenean from Chile. PMID:25033223

  19. Sulfide tolerance of four marine bioassay test species used to evaluate sediment toxicity

    SciTech Connect

    Knezovich, J.P.; Jelinski, J.A.; Anderson, S.L.; Steichen, D.J.

    1994-12-31

    Hydrogen sulfide is a potential positive interference associated with marine sediment toxicity tests. It is necessary, therefore, to evaluate the possible contribution of sulfides to sediment toxicity before toxicity attributed to anthropogenic contaminants can be determined. The sulfide tolerances of several marine bioassay test species (R. abronius, E. estuaris, M. edulis and S. purpuratus) commonly used in bulk sediment and porewater toxicity tests have not been reported. The authors used a flow-through exposure system to expose these organisms to constant concentrations of sulfide during 48-h exposure periods. Significant abnormalities occurred at 3 to 5 micromoles total sulfide per liter ({mu}M/L) in M. edulis embryos, and at 6{mu}M/L in S. purpuratus embryos. Survivorship of amphipods was significantly reduced at total sulfide concentrations of 45 and 126 {mu}M/L for R. abronius and E estuaris, respectively. These tolerances provide guidelines for evaluating the potential for sulfide toxicity prior to the initiation of toxicity tests. Because sulfide concentrations in bulk sediment and pore-water may exceed these values, care must be taken to ensure that sulfide toxicity is not contributing to toxicity observed when using these test organisms.

  20. Scanning electron microscopy study of adhesion in sea urchin blastulae. M.S. Thesis

    NASA Technical Reports Server (NTRS)

    Crowther, Susan D.

    1988-01-01

    The dissociation supernatant (DS) isolated by disaggregating Strongylocentrotus purpuratus blastulae in calcium- and magnesium-free seawater specifically promotes reaggregation of S. purpuratus blastula cells. The purpose of this study was to use scanning electron microscopy to examine the gross morphology of aggregates formed in the presence of DS to see if it resembles adhesion in partially dissociated blastulae. A new reaggregation procedure developed here, using large volumes of cell suspension and a large diameter of rotation, was utilized to obtain sufficient quantities of aggregates for scanning electron microscopy. The results indicate that aggregates formed in the presence of DS resemble partially dissociated intact embryos in terms of the direct cell-cell adhesion observed. DS did not cause aggregation to form as a result of the entrapment of cells in masses of extracellular material. These studies provide the groundwork for further studies using transmission electron microscopy to more precisely define the adhesive contacts made by cells in the presence of the putative adhesion molecules present in DS.

  1. Immunofluorescence localization of dissociation supernatant and extracellular matrix components in Lytechinus pictus sectioned embryos. M.S. Thesis

    NASA Technical Reports Server (NTRS)

    Garciaflack, Ana Leticia

    1988-01-01

    Indirect immunofluorescence was used to localize specific extracellular components in embryos of the sea urchin Lytechinus pictus. Hyalin and S2 (a group of components found in the disaggregation supernatant from Strongylocentrotus purpuratus blastulae) were uniformly present at all stages (unfertilized up to 32 hr) except hyalin could not be detected at the 12 hour early blastula stage. Laminin was found in 16 cell, 32 cell, 6 hour, 18 hour, 24 hour, and 32 hour stages, with especially bright fluorescence at 18 hours. Collagen I was present at all stages (freshly fertilized up to 32 hour) except little was detected at 12 hours. Fibronectin was uniformly present in blastocoelar fibers stained with anto-collagen I and anti-fibronectin. These results were compared with those for S. purpuratus to produce an overview of the localization of specific extracellular matrix components during development of two species of sea urchins. The results set the stage for future studies that will examine the function of these components at the various developmental stages.

  2. Kelp forest monitoring 1993 annual report. Channel Islands National Park. Final report

    SciTech Connect

    Kushner, D.; Walder, R.; Gorodezky, L.; Lerma, D.; Richards, D.

    1993-06-01

    The 1993 results of the Channel Islands National Park Kelp Forest Monitoring Project are described in this report. Population dynamics of 68 taxa or categories of algea, fish, and invertebrates were measured at 16 permanent sites around the five islands within the park. Survey techniques utilized SCUBA and surface-supplied-air, and included quadrats, band transects, random contacts, fish transects, video transects, size frequency measurements, artificial recruitment modules, and species list surveys. Temperature data was collected using Sea Data batheothermographs, and HOBOTEMP temperature loggers. Temperature loggers were installed at each of the sixteen sites. Size frequency measurements were taken from artifical recruitment modules at nine sites. In 1993, 13 sites had giant kelp, Macrocysts pyrifera, forests, one site was dominated by the aggregating red sea cucumber, pachythyone rubra, one site was dominated by red sea urchins, Strongylocentrotus franciscanus, and another by purple sea urchins, S. purpuratus. The 13 sites with kelp forests consisted of 10 mature and three young kelp forests. Wasting disease was observed in sea stars and wasting syndrome was apparent in sea urchins. Sea urchins wasting syndrome appears to have caused mass mortality of purple sea urchins, S. purpuratus, at two Santa Barbara Island sites.

  3. Rocky Intertidal Zonation Pattern in Antofagasta, Chile: Invasive Species and Shellfish Gathering

    PubMed Central

    Castilla, Juan Carlos; Manríquez, Patricio H.; Delgado, Alejandro; Ortiz, Verónica; Jara, María Elisa; Varas, Manuel

    2014-01-01

    Background Biological invasions affecting rocky intertidal zonation patterns, yield information on species interactions. In the Bay of Antofagasta, northern Chile, the non-indigenous tunicate Pyura praeputialis, originally from Australia, has invaded (in the past century or so) and monopolized a major portion of the mid-intertidal rocky shore, displacing upshore the native mussel Perumytilus purpuratus. In Antofagasta the tunicate is subjected to intensive exploitation. Monitoring protocols show that in the past 10 years Antofagasta's tunicate population has experienced a drastic decline, affecting the intertidal zonation pattern. Methodology/Principal Findings A 12.5 km of coastline, on the southern eastern shore of the Bay of Antofagasta, was studied. Eight sites were systematically (1993–1994) or sporadically (2003–2014) monitored for the seaward-shoreward expansion or reduction of the tunicate Pyura praeputialis, and native mussel and barnacle bands. A notable reduction in the mid-intertidal band of P. praeputialis and a seaward expansion of the mussel, Perumytilus purpuratus, and barnacle bands was observed. We suggest that the major cause for the decline in the tunicate is due to its intensive exploitation by rocky shore Pyura-gathers. The rate of extraction of tunicates by professional Pyura-gathers ranged between 256–740 tunicates hour−1. Between 2009–2014 the density of professional Pyura-gather ranged between 0.5–4.5 km−1 per low tide. Hence, 10 professional Pyura-gathers working 1 h for 10 low tides per month, during 6 months, will remove between 307–888 m2 of tunicates. A drastic decline in tunicate recruitment was observed and several P. praeputialis ecosystems services have been lost. Conclusion and Significance In Antofagasta, the continuous and intensive intertidal gathering of the invasive tunicate Pyura praeputialis, has caused a drastic reduction of its population modifying the zonation pattern. Thereby, native mussel Perumytilus

  4. Modulation of pumping rate by two species of marine bivalve molluscs in response to neurotransmitters: Comparison of in vitro and in vivo results.

    PubMed

    Frank, Dana M; Deaton, Lewis; Shumway, Sandra E; Holohan, Bridget A; Ward, J Evan

    2015-07-01

    Most studies regarding the neuroanatomy and neurophysiology of molluscan ctenidia have focused on isolated ctenidial tissue preparations. This study investigated how bivalve molluscs modulate their feeding rates by examining the effects of a variety of neurotransmitters, including serotonin, dopamine, and the dopamine agonist apomorphine on both isolated ctenidial tissue and in intact members of two commercially important bivalve species: the blue mussel, Mytilus edulis; and the bay scallop Argopecten irradians. In particular, we examined the effect of changes in: 1) beat of the lateral cilia (in vitro), 2) distance between ctenidial filaments and/or plicae (in vivo), and 3) diameter of the siphonal openings (in vivo) on alteration of bulk water flow through the mantle cavity. Important differences were found between isolated tissue and whole animals, and between species. Drugs that stimulated ciliary beat in vitro did not increase water processing rate in vivo. None of the treatments increased water flow through the mantle cavity of intact animals. Results suggest that A. irradians was primarily modulating lateral ciliary activity, while M. edulis appeared to have a number of ways to control water processing activity, signifying that the two species may have different compensatory and regulatory mechanisms controlling feeding activity. PMID:25847101

  5. Effects of Elevated Temperature and Carbon Dioxide on the Growth and Survival of Larvae and Juveniles of Three Species of Northwest Atlantic Bivalves

    PubMed Central

    Talmage, Stephanie C.; Gobler, Christopher J.

    2011-01-01

    Rising CO2 concentrations and water temperatures this century are likely to have transformative effects on many coastal marine organisms. Here, we compared the responses of two life history stages (larval, juvenile) of three species of calcifying bivalves (Mercenaria mercenaria, Crassostrea virginica, and Argopecten irradians) to temperatures (24 and 28°C) and CO2 concentrations (∼250, 390, and 750 ppm) representative of past, present, and future summer conditions in temperate estuaries. Results demonstrated that increases in temperature and CO2 each significantly depressed survival, development, growth, and lipid synthesis of M. mercenaria and A. irradians larvae and that the effects were additive. Juvenile M. mercenaria and A. irradians were negatively impacted by higher temperatures while C. virginica juveniles were not. C. virginica and A. irradians juveniles were negatively affected by higher CO2 concentrations, while M. mercenaria was not. Larvae were substantially more vulnerable to elevated CO2 than juvenile stages. These findings suggest that current and future increases in temperature and CO2 are likely to have negative consequences for coastal bivalve populations. PMID:22066018

  6. Effects of past, present, and future ocean carbon dioxide concentrations on the growth and survival of larval shellfish

    PubMed Central

    Gobler, Christopher J.

    2010-01-01

    The combustion of fossil fuels has enriched levels of CO2 in the world’s oceans and decreased ocean pH. Although the continuation of these processes may alter the growth, survival, and diversity of marine organisms that synthesize CaCO3 shells, the effects of ocean acidification since the dawn of the industrial revolution are not clear. Here we present experiments that examined the effects of the ocean’s past, present, and future (21st and 22nd centuries) CO2 concentrations on the growth, survival, and condition of larvae of two species of commercially and ecologically valuable bivalve shellfish (Mercenaria mercenaria and Argopecten irradians). Larvae grown under near preindustrial CO2 concentrations (250 ppm) displayed significantly faster growth and metamorphosis as well as higher survival and lipid accumulation rates compared with individuals reared under modern day CO2 levels. Bivalves grown under near preindustrial CO2 levels displayed thicker, more robust shells than individuals grown at present CO2 concentrations, whereas bivalves exposed to CO2 levels expected later this century had shells that were malformed and eroded. These results suggest that the ocean acidification that has occurred during the past two centuries may be inhibiting the development and survival of larval shellfish and contributing to global declines of some bivalve populations. PMID:20855590

  7. Selected alternatives to conventional chlorination. Final report

    SciTech Connect

    Garey, J.F.

    1980-10-01

    This study was jointly funded by EPRI and five electric utility companies in New England (New England Power, Northeast Utilities, United Illuminating, Vermont Yankee Nuclear, and Public Service of New Hampshire). Previous investigations had identified three major areas for further study: continuous low-level chlorination, dechlorination, and condenser biofouling control. Continuous low-level chlorination, studied at two locations, one on open coastal water and the other in an industrialized estuarine area, showed that 0.1 ppM total residual oxidant (TRO) prevented attachment of the blue mussel (Mytilus edulis) to concrete surfaces. Chronic bioassays showed that 0.075 ppM TRO reduced biofouling by indigenous organisms; 0.1 ppM TRO slightly increased mortalities of the Atlantic silversides (Menidia menidia) but had no effect on the American oyster (Crassostrea virginica). Dechlorination investigations showed that threespine sticklebacks (Gasterosteus aculeatus), Atlantic silversides (Menidia menidia), larval bay scallops (Argopecten irradians), and the copepod Acartia tonsa exposed to water chlorinated to 0.5 ppM TRO for 10, 100, and 1000 seconds, followed by dechlorination with sodium thiosulfate, all suffered significant toxic effects. Condenser tube biofouling studies showed that there was a strong correlation between condenser performance and condenser tube biofouling; biofilm induction varied inversely with ambient water temperature, but orientation of the tubes had no effect on biofilm formation; and all chemicals tested (mono-, di-, and trisodium phosphate; Polident; and TRO at 0.1 ppM) reduced but did not remove biofilms.

  8. Composition, abundance, biomass, and production of macrofauna in a New England estuary: comparisons among eelgrass meadows and other nursery habitats

    USGS Publications Warehouse

    Heck, K.L., Jr.; Able, K.W.; Roman, C.T.; Fahay, M.P.

    1995-01-01

    Quantitative suction sampling was used to characterize and compare the species composition, abundance, biomass, and secondary production of macrofauna inhabiting intertidal mudflat and sandflat, eelgrass meadow, and saltmarshpool habitats in the Nauset Marsh complex, Cape Cod, Massachusetts (USA). Species richness and abundance were often greatest in eelgrass habitat, as was macroinvertebrate biomass and production. Most striking was the five to fifteen times greater rate of annual macrofaunal production in eelgrass habitat than elsewhere, with values ranging from approximately 23139 g AFDW m super(2) yr super(1). The marsh pool containing widgeon grass (Ruppia maritima) supported surprisingly low numbers of macroinvertebrates, probably due to stressfully low dissolved oxygen levels at night during the summer. Two species of macroinvertebrates, blue mussels (Mytilus edulis) and to a lesser extent bay scallops (Argopecten irradians), used eelgrass as 'nursery habitat.' Calculations showed that macroinvertebrate production is proportionally much greater than the amount of primary production attributable to eelgrass in the Nauset Marsh system, and that dramatic changes at all trophic levels could be expected if large changes in seagrass abundance should occur. This work further underscores the extraordinarily large impact that seagrass can have on both the structure and function of estuarine ecosystems.

  9. Cloning of a big defensin gene and its response to Vibrio parahaemolyticus challenge in the noble scallop Chlamys nobilis (Bivalve: Pectinidae).

    PubMed

    Yang, Jianqing; Luo, Jiafu; Zheng, Huaiping; Lu, Yeqing; Zhang, Hongkuan

    2016-09-01

    The noble scallop Chlamys nobilis has been an important marine cultured bivalve in the Southern Sea of China for decades. However, large-scale mortality events often occurred during the scallop' cultivation. As one of AMPs (antimicrobial peptides), big defensin is an important component of the innate immunity against pathogenic microorganisms in invertebrates. In order to investigate whether the big defensin can play a role in the immune defense against pathogenic microorganisms in noble scallop, a big defensin gene from the hemocytes of Chlamys nobilis (CnBD) was cloned, and the mRNA level was measured after an acute Vibrio parahaemolyticus challenge of 36 h. The CnBD cDNA contains an open reading frame (ORF) of 381 bp encoding a peptide of 126 amino acids residues. The deduce amino acid sequence of CnBD shows a high similarity with that from Argopecten irradians and displays common features of big defensin, indicating that CnBD is a new member of the big defensin family. Compared with the control group, the relative mRNA level of CnBD was significantly up-regulated at 3, 24 and 36 h. The present result indicated that CnBD played an immune role against bacterial infection in noble scallop. PMID:27474446

  10. Kelp forest monitoring 1994 annual report. Channel Islands National Park. Final report

    SciTech Connect

    Kushner, D.; Lerma, D.; Richards, D.

    1994-12-31

    The 1994 results of the Channel Islands Natonal Park Kelp Forest Monitoring Project are described in this report. Population dynamics of 68 taxa or categories of algae, fish, and invertebrates were measured at 16 permanent sites around the five islands within the park. Survey techniques utilized SCUBA and surface-supplied-air, and included quadrants, band transects, random point contacts, fish transects, video transects, size frequency measurements, artificial recruitment modules, and species list surveys. Temperature data was collected using temperature loggers deployed at each of the sixteen sites. Size frequency measurements were taken from artificial recruitment modules at ten sites. In 1994, 13 sites had giant kelp, Macrocystis pyrifera, forests, one site was dominated by the aggregating red sea cucumber, Pachythyone rubra, one site was dominated by red sea urchins, Strongylocentrotus francisanus, and another by purple sea urchins, S. purpuratus. Wasting disease was observed in sea stars and wasting syndrome was apparent in sea urchins.

  11. Poster: the sea urchin.

    PubMed

    2006-11-10

    The announcement of the sequencing of the genome of the sea urchin Strongylocentrotus purpuratus is the latest glowing chapter in the long history of contributions from sea urchin researchers to fundamental principles of cell biology, developmental biology, and gene regulation. A poster created for the print Science presents a timeline of the past and a glimpse into the next stage of research that is being opened up through genomic insights; a PDF of that poster is available for our subscribers. We have also created, for all site visitors, an interactive version of the poster, including additional images, video, and Web resources. Interactive poster (free; requires Flash 8 or higher). PDF of print poster (subscription required). PMID:17095688

  12. Proteome-wide dataset supporting the study of ancient metazoan macromolecular complexes.

    PubMed

    Phanse, Sadhna; Wan, Cuihong; Borgeson, Blake; Tu, Fan; Drew, Kevin; Clark, Greg; Xiong, Xuejian; Kagan, Olga; Kwan, Julian; Bezginov, Alexandr; Chessman, Kyle; Pal, Swati; Cromar, Graham; Papoulas, Ophelia; Ni, Zuyao; Boutz, Daniel R; Stoilova, Snejana; Havugimana, Pierre C; Guo, Xinghua; Malty, Ramy H; Sarov, Mihail; Greenblatt, Jack; Babu, Mohan; Derry, W Brent; Tillier, Elisabeth R; Wallingford, John B; Parkinson, John; Marcotte, Edward M; Emili, Andrew

    2016-03-01

    Our analysis examines the conservation of multiprotein complexes among metazoa through use of high resolution biochemical fractionation and precision mass spectrometry applied to soluble cell extracts from 5 representative model organisms Caenorhabditis elegans, Drosophila melanogaster, Mus musculus, Strongylocentrotus purpuratus, and Homo sapiens. The interaction network obtained from the data was validated globally in 4 distant species (Xenopus laevis, Nematostella vectensis, Dictyostelium discoideum, Saccharomyces cerevisiae) and locally by targeted affinity-purification experiments. Here we provide details of our massive set of supporting biochemical fractionation data available via ProteomeXchange (PXD002319-PXD002328), PPIs via BioGRID (185267); and interaction network projections via (http://metazoa.med.utoronto.ca) made fully accessible to allow further exploration. The datasets here are related to the research article on metazoan macromolecular complexes in Nature [1]. PMID:26870755

  13. SEM and x-ray microanalysis of cellular differentiation in Sea Urchin Embryos: a frozen hydrated study

    SciTech Connect

    Klein, S.B.

    1985-12-01

    Quantitative studies of major chemical element distribution among individual differentiating cells were attempted using scanning electron microscopy. Frozen hydrated embryos of the sea urchin Strongelocentrotus purpuratus were examined at three stages: blastula, mesenchyme blastula, and early gastrula. The blastocoel matrix contained large beads of approximately 1 ..mu..m diameter. The cells of the archenteron lacked well defined cell boundaries. Characteristic levels of beam damage and charging provided structural information. The primary mesenchyme cells within the blastocoel were particularly susceptible to both effects. Damaging effects were noted in material stored in liquid nitrogen longer than three months. Ice crystal growth, shrinkage, elemental shift, density changes and charge accumulation may take place in these stored specimens. 151 refs., 50 figs., 3 tabs.

  14. Spermidine is bound to a unique protein in early sea urchin embryos

    SciTech Connect

    Canellakis, Z.N.; Bondy, P.K.; Infante, A.A.

    1985-11-01

    Spermidine is rapidly taken up and becomes bound to protein during the very early hours of sea urchin embryogenesis. During the first 6 hr after fertilization of freshly obtained sea urchin eggs (Strongylocentrotus purpuratus), which are incubated in the presence of exogenous (/sup 3/H)-spermidine, up to 7% of the total cell-associated spermidine appears uniquely as spermidine bound in macromolecular form. This unique protein containing spermidine migrates as a single radioactive band in gel electrophoresis. It has a Mr of approximately equal to 30,000 and is readily distinguishable from the protein initiation factor eIF-4D, which has a Mr of 18,000, the only other identifiable protein known to date to be posttranslationally modified by polyamines.

  15. Expression, purification, crystallization and preliminary X-ray analysis of the olfactomedin domain from the sea urchin cell-adhesion protein amassin

    SciTech Connect

    Hillier, Brian J.; Sundaresan, Vidyasankar; Stout, C. David; Vacquier, Victor D.

    2006-01-01

    The olfactomedin (OLF) domain from the sea urchin cell-adhesion protein amassin has been crystallized. A native data set extending to 2.7 Å has been collected using an in-house X-ray source. A family of animal proteins is emerging which contain a conserved protein motif known as an olfactomedin (OLF) domain. Novel extracellular protein–protein interactions occur through this domain. The OLF-family member amassin, from the sea urchin Strongylocentrotus purpuratus, has previously been identified to mediate a rapid cell-adhesion event resulting in a large aggregation of coelomocytes, the circulating immune cells. In this work, heterologous expression and purification of the OLF domain from amassin was carried out and initial crystallization trials were performed. A native data set has been collected, extending to 2.7 Å under preliminary cryoconditions, using an in-house generator. This work leads the way to the determination of the first structure of an OLF domain.

  16. Influence of 60-Hz magnetic fields on sea urchin development

    SciTech Connect

    Zimmerman, S.; Zimmerman, A.M.; Winters, W.D.; Cameron, I.L. )

    1990-01-01

    Continuous exposure of sea urchin (Strongylocentrotus purpuratus) embryos at 18 degrees C to a cyclic 60-Hz magnetic field at 0.1 mT rms beginning 4 min after insemination caused a significant developmental delay during the subsequent 23 hours. No delay in development was recorded for periods up to 18 hours after fertilization. At 18 h, most embryos were in the mesenchyme blastula stage. At 23 h, most control embryos were in mid-gastrula whereas most magnetic-field-exposed embryos were in the early gastrula stage. Thus an estimated 1-h delay occurred between these developmental stages. The results are discussed in terms of possible magnetic-field modification of transcription as well as interference with cell migration during gastrulation. The present study extends and supports the growing body of information about potential effects of exposures to extremely-low-frequency (ELF) magnetic fields on developing organisms.

  17. A partial skeletal proteome of the brittle star Ophiocoma wendtii

    NASA Astrophysics Data System (ADS)

    Seaver, Ryan W.

    The formation of mineralized tissue was critical to the evolution and diversification of metazoans and remains functionally significant in most animal lineages. Of special importance is the protein found occluded within the mineral matrix, which facilitates the process of biomineralization and modulates the final mineral structure. These skeletal matrix proteins have well been described in several species, including the sea urchin Stronglyocentrotus purpuratus, an important model organism. Biomineralization research is limited in other echinoderm classes. This research encompasses the first description of mineral matrix proteins in a member of the echinoderm class Ophiuroidea. This work describes the skeletal matrix proteins of the brittle star Ophiocoma wendtii using bioinformatic and proteomic techniques. General characteristics of matrix protein are described and a number of candidate biomineralization related genes have been identified, cloned, and sequenced. The unique evolutionary and biochemical properties of brittle star skeletal matrix proteins are also described.

  18. Applied DC magnetic fields cause alterations in the time of cell divisions and developmental abnormalities in early sea urchin embryos

    SciTech Connect

    Levin, M.; Ernst, S.G.

    1997-05-01

    Most work on magnetic field effects focuses on AC fields. The present study demonstrates that exposure to medium-strength (10 mT--0.1 T) static magnetic fields can alter the early embryonic development of two species of sea urchin embryos. Batches of fertilized eggs from two species of urchin were exposed to fields produced by permanent magnets. Samples of the continuous cultures were scored for the timing of the first two cell divisions, time of hatching, and incidence of exogastrulation. It was found that static fields delay the onset of mitosis in both species by an amount dependent on the exposure timing relative to fertilization. The exposure time that caused the maximum effect differed between the two species. Thirty millitesla fields, but not 15 mT fields, caused an eightfold increase in the incidence of exogastrulation in Lytechinus pictus, whereas neither of these fields produced exogastrulation in Strongylocentrotus purpuratus.

  19. Assessment of chronic toxicity of petroleum and produced water components to marine organisms. Final technical summary

    SciTech Connect

    Cherr, G.N.; Higashi, R.M.; Shenker, J.M.

    1993-05-31

    The objectives of the report were: (1) to determine the effects of produced water exposure in early life stages of marine plants and animals, at the cellular, subcellular, and physiological levels; (2) to determine the effects of produced water exposure on reproduction in marine organisms; and (3) to develop non-invasive approaches for assessing reproductive impairment. The effects of produced water (PW) was assessed on development in three ecologically and economically important species, the purple sea urchin (Strongylocentrotus purpuratus), the giant kelp (macrocystis pyrifera), and tsahe California mussel (Mytilus califonrnianus). To determine the basis for effects of PW on these developing organisms, some fundamental studies were prerequisite. Furthermore, eggs and embryos from adults which were outplanted near the discharge were also studied. Finally, the biochemical response of embryos to PW was also defined.

  20. Post-translational regulation by gustavus contributes to selective Vasa protein accumulation in multipotent cells during embryogenesis

    PubMed Central

    Gustafson, Eric A.; Yajima, Mamiko; Juliano, Celina E.; Wessel, Gary M.

    2010-01-01

    Vasa is a broadly conserved DEAD-box RNA helicase associated with germ line development and is expressed in multipotent cells in many animals. During embryonic development of the sea urchin Strongylocentrotus purpuratus, Vasa protein is enriched in the small micromeres despite a uniform distribution of vasa transcript. Here we show that the Vasa coding region is sufficient for its selective enrichment and find that gustavus, the B30.2/SPRY and SOCS box domain gene, contributes to this phenomenon. In vitro binding analyses show that Gustavus binds the N-terminal and DEAD-box portions of Vasa protein independently. A knockdown of Gustavus protein reduces both Vasa protein abundance and its propensity for accumulation in the small micromeres, whereas overexpression of the Vasa-interacting domain of Gustavus (GusΔSOCS) results in Vasa protein accumulation throughout the embryo. We propose that Gustavus has a conserved, positive regulatory role in Vasa protein accumulation during embryonic development. PMID:21035437

  1. Proteome-wide dataset supporting the study of ancient metazoan macromolecular complexes

    PubMed Central

    Phanse, Sadhna; Wan, Cuihong; Borgeson, Blake; Tu, Fan; Drew, Kevin; Clark, Greg; Xiong, Xuejian; Kagan, Olga; Kwan, Julian; Bezginov, Alexandr; Chessman, Kyle; Pal, Swati; Cromar, Graham; Papoulas, Ophelia; Ni, Zuyao; Boutz, Daniel R.; Stoilova, Snejana; Havugimana, Pierre C.; Guo, Xinghua; Malty, Ramy H.; Sarov, Mihail; Greenblatt, Jack; Babu, Mohan; Derry, W. Brent; Tillier, Elisabeth R.; Wallingford, John B.; Parkinson, John; Marcotte, Edward M.; Emili, Andrew

    2015-01-01

    Our analysis examines the conservation of multiprotein complexes among metazoa through use of high resolution biochemical fractionation and precision mass spectrometry applied to soluble cell extracts from 5 representative model organisms Caenorhabditis elegans, Drosophila melanogaster, Mus musculus, Strongylocentrotus purpuratus, and Homo sapiens. The interaction network obtained from the data was validated globally in 4 distant species (Xenopus laevis, Nematostella vectensis, Dictyostelium discoideum, Saccharomyces cerevisiae) and locally by targeted affinity-purification experiments. Here we provide details of our massive set of supporting biochemical fractionation data available via ProteomeXchange (PXD002319-PXD002328), PPIs via BioGRID (185267); and interaction network projections via (http://metazoa.med.utoronto.ca) made fully accessible to allow further exploration. The datasets here are related to the research article on metazoan macromolecular complexes in Nature [1]. PMID:26870755

  2. Turbidity as a method of preparing sperm dilutions in the echinoid sperm/egg bioassay

    SciTech Connect

    Hall, T.J.; Haley, R.K.; Battan, K.J. )

    1993-11-01

    The use of turbidimeter for preparing sperm dilutions used in the echinoid sperm/egg bioassay was evaluated. Regression analyses of the relationship between sperm density and turbidity for the sea urchins Strongylocentrotus purpuratus and Strongylocentrotus droebachiensis and the sand dollar Dendraster excentricus indicated that although there were slope differences for each species, each coefficient of determination was highly significant. For Dendraster excentricus, triplicate hemacytometer counts over a range of turbidities as well as repeated preparations of a single sperm turbidity indicated similar variability for each. The use of the turbidimeter has time-saving advantages over conventional hemacytometer methods without sacrificing precision. Sperm dilutions can be prepared rapidly, minimizing seawater sperm preactivation before test initiation, and may therefore contribute to increased test precision.

  3. Substituting mouse transcription factor Pou4f2 with a sea urchin orthologue restores retinal ganglion cell development

    PubMed Central

    Mocko-Strand, Julie A.; Wang, Jing; Ullrich-Lüter, Esther; Pan, Ping; Wang, Steven W.; Arnone, Maria Ina; Frishman, Laura J.; Klein, William H.

    2016-01-01

    Pou domain transcription factor Pou4f2 is essential for the development of retinal ganglion cells (RGCs) in the vertebrate retina. A distant orthologue of Pou4f2 exists in the genome of the sea urchin (class Echinoidea) Strongylocentrotus purpuratus (SpPou4f1/2), yet the photosensory structure of sea urchins is strikingly different from that of the mammalian retina. Sea urchins have no obvious eyes, but have photoreceptors clustered around their tube feet disc. The mechanisms that are associated with the development and function of photoreception in sea urchins are largely unexplored. As an initial approach to better understand the sea urchin photosensory structure and relate it to the mammalian retina, we asked whether SpPou4f1/2 could support RGC development in the absence of Pou4f2. To answer this question, we replaced genomic Pou4f2 with an SpPou4f1/2 cDNA. In Pou4f2-null mice, retinas expressing SpPou4f1/2 were outwardly identical to those of wild-type mice. SpPou4f1/2 retinas exhibited dark-adapted electroretinogram scotopic threshold responses, indicating functionally active RGCs. During retinal development, SpPou4f1/2 activated RGC-specific genes and in S. purpuratus, SpPou4f2 was expressed in photoreceptor cells of tube feet in a pattern distinct from Opsin4 and Pax6. Our results suggest that SpPou4f1/2 and Pou4f2 share conserved components of a gene network for photosensory development and they maintain their conserved intrinsic functions despite vast morphological differences in mouse and sea urchin photosensory structures. PMID:26962139

  4. Reconstructing SALMFamide Neuropeptide Precursor Evolution in the Phylum Echinodermata: Ophiuroid and Crinoid Sequence Data Provide New Insights

    PubMed Central

    Elphick, Maurice R.; Semmens, Dean C.; Blowes, Liisa M.; Levine, Judith; Lowe, Christopher J.; Arnone, Maria I.; Clark, Melody S.

    2015-01-01

    The SALMFamides are a family of neuropeptides that act as muscle relaxants in echinoderms. Analysis of genome/transcriptome sequence data from the sea urchin Strongylocentrotus purpuratus (Echinoidea), the sea cucumber Apostichopus japonicus (Holothuroidea), and the starfish Patiria miniata (Asteroidea) reveals that in each species there are two types of SALMFamide precursor: an L-type precursor comprising peptides with a C-terminal LxFamide-type motif and an F-type precursor solely or largely comprising peptides with a C-terminal FxFamide-type motif. Here, we have identified transcripts encoding SALMFamide precursors in the brittle star Ophionotus victoriae (Ophiuroidea) and the feather star Antedon mediterranea (Crinoidea). We have also identified SALMFamide precursors in other species belonging to each of the five echinoderm classes. As in S. purpuratus, A. japonicus, and P. miniata, in O. victoriae there is one L-type precursor and one F-type precursor. However, in A. mediterranea only a single SALMFamide precursor was found, comprising two peptides with a LxFamide-type motif, one with a FxFamide-type motif, five with a FxLamide-type motif, and four with a LxLamide-type motif. As crinoids are basal to the Echinozoa (Holothuroidea + Echinoidea) and Asterozoa (Asteroidea + Ophiuroidea) in echinoderm phylogeny, one model of SALMFamide precursor evolution would be that ancestrally there was a single SALMFamide gene encoding a variety of SALMFamides (as in crinoids), which duplicated in a common ancestor of the Echinozoa and Asterozoa and then specialized to encode L-type SALMFamides or F-type SALMFamides. Alternatively, a second SALMFamide precursor may remain to be discovered or may have been lost in crinoids. Further insights will be obtained if SALMFamide receptors are identified, which would provide a molecular basis for experimental analysis of the functional significance of the “cocktails” of SALMFamides that exist in echinoderms. PMID:25699014

  5. Reconstructing SALMFamide Neuropeptide Precursor Evolution in the Phylum Echinodermata: Ophiuroid and Crinoid Sequence Data Provide New Insights.

    PubMed

    Elphick, Maurice R; Semmens, Dean C; Blowes, Liisa M; Levine, Judith; Lowe, Christopher J; Arnone, Maria I; Clark, Melody S

    2015-01-01

    The SALMFamides are a family of neuropeptides that act as muscle relaxants in echinoderms. Analysis of genome/transcriptome sequence data from the sea urchin Strongylocentrotus purpuratus (Echinoidea), the sea cucumber Apostichopus japonicus (Holothuroidea), and the starfish Patiria miniata (Asteroidea) reveals that in each species there are two types of SALMFamide precursor: an L-type precursor comprising peptides with a C-terminal LxFamide-type motif and an F-type precursor solely or largely comprising peptides with a C-terminal FxFamide-type motif. Here, we have identified transcripts encoding SALMFamide precursors in the brittle star Ophionotus victoriae (Ophiuroidea) and the feather star Antedon mediterranea (Crinoidea). We have also identified SALMFamide precursors in other species belonging to each of the five echinoderm classes. As in S. purpuratus, A. japonicus, and P. miniata, in O. victoriae there is one L-type precursor and one F-type precursor. However, in A. mediterranea only a single SALMFamide precursor was found, comprising two peptides with a LxFamide-type motif, one with a FxFamide-type motif, five with a FxLamide-type motif, and four with a LxLamide-type motif. As crinoids are basal to the Echinozoa (Holothuroidea + Echinoidea) and Asterozoa (Asteroidea + Ophiuroidea) in echinoderm phylogeny, one model of SALMFamide precursor evolution would be that ancestrally there was a single SALMFamide gene encoding a variety of SALMFamides (as in crinoids), which duplicated in a common ancestor of the Echinozoa and Asterozoa and then specialized to encode L-type SALMFamides or F-type SALMFamides. Alternatively, a second SALMFamide precursor may remain to be discovered or may have been lost in crinoids. Further insights will be obtained if SALMFamide receptors are identified, which would provide a molecular basis for experimental analysis of the functional significance of the "cocktails" of SALMFamides that exist in echinoderms. PMID:25699014

  6. Molecular cloning of hsf1 and hsbp1 cDNAs, and the expression of hsf1, hsbp1 and hsp70 under heat stress in the sea cucumber Apostichopus japonicus.

    PubMed

    Xu, Dongxue; Sun, Lina; Liu, Shilin; Zhang, Libin; Yang, Hongsheng

    2016-08-01

    The heat shock response (HSR) is known for the elevated synthesis of heat shock proteins (HSPs) under heat stress, which is mediated primarily by heat shock factor 1 (HSF1). Heat shock factor binding protein 1 (HSBP1) and feedback control of heat shock protein 70 (HSP70) are major regulators of the activity of HSF1. We obtained full-length cDNA of genes hsf1 and hsbp1 in the sea cucumber Apostichopus japonicus, which are the second available for echinoderm (after Strongylocentrotus purpuratus), and the first available for holothurian. The full-length cDNA of hsf1 was 2208bp, containing a 1326bp open reading frame encoding 441 amino acids. The full-length cDNA of hsbp1 was 2850bp, containing a 225bp open reading frame encoding 74 amino acids. The similarities of A. japonicus HSF1 with other species are low, and much higher similarity identities of A. japonicus HSBP1 were shared. Phylogenetic trees showed that A. japonicus HSF1 and HSBP1 were clustered with sequences from S. purpuratus, and fell into distinct clades with sequences from mollusca, arthropoda and vertebrata. Analysis by real-time PCR showed hsf1 and hsbp1 mRNA was expressed constitutively in all tissues examined. The expression of hsf1, hsbp1 and hsp70 in the intestine at 26°C was time-dependent. The results of this study might provide new insights into the regulation of heat shock response in this species. PMID:26952354

  7. Identification, Modeling and Ligand Affinity of Early Deuterostome CYP51s, and Functional Characterization of Recombinant Zebrafish Sterol 14α-Demethylase

    PubMed Central

    Morrison, Ann Michelle Stanley; Goldstone, Jared V.; Lamb, David C.; Kubota, Akira; Lemaire, Benjamin; Stegeman, John. J.

    2014-01-01

    Background Sterol 14α-demethylase (cytochrome P450 51, CYP51, P45014DM) is a microsomal enzyme that in eukaryotes catalyzes formation of sterols essential for cell membrane function and as precursors in biosynthesis of steroid hormones. Functional properties of CYP51s are unknown in non-mammalian deuterostomes. Methods PCR-cloning and sequencing and computational analyses (homology modeling and docking) addressed CYP51 in zebrafish Danio rerio, the reef fish sergeant major Abudefduf saxatilis, and the sea urchin Strongylocentrotus purpuratus. Following N-terminal amino acid modification, zebrafish CYP51 was expressed in Escherichia coli, and lanosterol 14α-demethylase activity and azole inhibition of CYP51 activity were characterized using GC/MS. Results Molecular phylogeny positioned S. purpuratus CYP51 at the base of the deuterostome clade. In zebrafish, CYP51 is expressed in all organs examined, most strongly in intestine. The recombinant protein bound lanosterol and catalyzed 14α-demethylase activity, at 3.2 nmol/min/nmol CYP51. The binding of azoles to zebrafish CYP51 gave KS (dissociation constant) values of 0.26 μM for ketoconazole and 0.64 μM for propiconazole. Displacement of carbon monoxide also indicated zebrafish CYP51 has greater affinity for ketoconazole. Docking to homology models showed that lanosterol docks in fish and sea urchin CYP51s with an orientation essentially the same as in mammalian CYP51. Docking of ketoconazole indicates it would inhibit fish and sea urchin CYP51s. Conclusions Biochemical and computational analyses are consistent with lanosterol being a substrate for early deuterostome CYP51s. General Significance The results expand the phylogenetic view of animal CYP51, with evolutionary, environmental and therapeutic implications. PMID:24361620

  8. Cell-surface proteoglycan in sea urchin primary mesenchyme cell migration

    SciTech Connect

    Lane, M.C.

    1989-01-01

    Early in the development of the sea urchin embryo, the primary mesenchyme cells (PMC) migrate along the basal lamina of the blastocoel. Migration is inhibited in L. pictus embryos cultured in sulfate-free seawater and in S. purpuratus embryos exposed to exogenous {beta}-D-xylosides. An in vitro assay was developed to test the migratory capacity of normal PMC on normal and treated blastocoelic matrix. Sulfate deprivation and exposure to exogenous xyloside render PMC nonmotile on either matrix. Materials removed from the surface of normal PMC by treatment with 1 M urea restored migratory ability to defective cells, whereas a similar preparation isolated from the surface of epithelial cells at the same stage did not. Migration also resumed when cells were removed from the xyloside or returned to normal seawater. The urea extract was partially purified and characterized by radiolabeling, gel electrophoresis, fluorography, ion exchange chromatography, and western blotting. The PMC synthesize a large chondroitin sulfate/dermatan sulfate proteoglycan that is present in an active fraction isolated by chromatography. Chondroitinase ABC digestion of live cells blocked migration reversibly, further supporting the identification of the chondroitin sulfate/dermatan sulfate proteoglycan as the active component in the urea extract. Much of the incorporated sulfate was distributed along the filopodia in {sup 35}SO{sub 4}-labelled PMC by autoradiography. The morphology of normal and treated S. purpuratus PMC was examined by scanning electron microscopy, and differences in spreading, particularly of the extensive filopodia present on the cells, was observed. A model for the role of the chondroitin sulfate/dermatan sulfate proteoglycan in cell detachment during migration is proposed.

  9. Substituting mouse transcription factor Pou4f2 with a sea urchin orthologue restores retinal ganglion cell development.

    PubMed

    Mao, Chai-An; Agca, Cavit; Mocko-Strand, Julie A; Wang, Jing; Ullrich-Lüter, Esther; Pan, Ping; Wang, Steven W; Arnone, Maria Ina; Frishman, Laura J; Klein, William H

    2016-03-16

    Pou domain transcription factor Pou4f2 is essential for the development of retinal ganglion cells (RGCs) in the vertebrate retina. A distant orthologue of Pou4f2 exists in the genome of the sea urchin (class Echinoidea) Strongylocentrotus purpuratus (SpPou4f1/2), yet the photosensory structure of sea urchins is strikingly different from that of the mammalian retina. Sea urchins have no obvious eyes, but have photoreceptors clustered around their tube feet disc. The mechanisms that are associated with the development and function of photoreception in sea urchins are largely unexplored. As an initial approach to better understand the sea urchin photosensory structure and relate it to the mammalian retina, we asked whether SpPou4f1/2 could support RGC development in the absence of Pou4f2. To answer this question, we replaced genomic Pou4f2 with an SpPou4f1/2 cDNA. In Pou4f2-null mice, retinas expressing SpPou4f1/2 were outwardly identical to those of wild-type mice. SpPou4f1/2 retinas exhibited dark-adapted electroretinogram scotopic threshold responses, indicating functionally active RGCs. During retinal development, SpPou4f1/2 activated RGC-specific genes and in S. purpuratus, SpPou4f2 was expressed in photoreceptor cells of tube feet in a pattern distinct from Opsin4 and Pax6. Our results suggest that SpPou4f1/2 and Pou4f2 share conserved components of a gene network for photosensory development and they maintain their conserved intrinsic functions despite vast morphological differences in mouse and sea urchin photosensory structures. PMID:26962139

  10. Conservation of Endo16 expression in sea urchins despite evolutionary divergence in both cis and trans-acting components of transcriptional regulation

    NASA Technical Reports Server (NTRS)

    Romano, Laura A.; Wray, Gregory A.

    2003-01-01

    Evolutionary changes in transcriptional regulation undoubtedly play an important role in creating morphological diversity. However, there is little information about the evolutionary dynamics of cis-regulatory sequences. This study examines the functional consequence of evolutionary changes in the Endo16 promoter of sea urchins. The Endo16 gene encodes a large extracellular protein that is expressed in the endoderm and may play a role in cell adhesion. Its promoter has been characterized in exceptional detail in the purple sea urchin, Strongylocentrotus purpuratus. We have characterized the structure and function of the Endo16 promoter from a second sea urchin species, Lytechinus variegatus. The Endo16 promoter sequences have evolved in a strongly mosaic manner since these species diverged approximately 35 million years ago: the most proximal region (module A) is conserved, but the remaining modules (B-G) are unalignable. Despite extensive divergence in promoter sequences, the pattern of Endo16 transcription is largely conserved during embryonic and larval development. Transient expression assays demonstrate that 2.2 kb of upstream sequence in either species is sufficient to drive GFP reporter expression that correctly mimics this pattern of Endo16 transcription. Reciprocal cross-species transient expression assays imply that changes have also evolved in the set of transcription factors that interact with the Endo16 promoter. Taken together, these results suggest that stabilizing selection on the transcriptional output may have operated to maintain a similar pattern of Endo16 expression in S. purpuratus and L. variegatus, despite dramatic divergence in promoter sequence and mechanisms of transcriptional regulation.

  11. Comparison of the receptor FGFRL1 from sea urchins and humans illustrates evolution of a zinc binding motif in the intracellular domain

    PubMed Central

    2009-01-01

    Background FGFRL1, the gene for the fifth member of the fibroblast growth factor receptor (FGFR) family, is found in all vertebrates from fish to man and in the cephalochordate amphioxus. Since it does not occur in more distantly related invertebrates such as insects and nematodes, we have speculated that FGFRL1 might have evolved just before branching of the vertebrate lineage from the other invertebrates (Beyeler and Trueb, 2006). Results We identified the gene for FGFRL1 also in the sea urchin Strongylocentrotus purpuratus and cloned its mRNA. The deduced amino acid sequence shares 62% sequence similarity with the human protein and shows conservation of all disulfides and N-linked carbohydrate attachment sites. Similar to the human protein, the S. purpuratus protein contains a histidine-rich motif at the C-terminus, but this motif is much shorter than the human counterpart. To analyze the function of the novel motif, recombinant fusion proteins were prepared in a bacterial expression system. The human fusion protein bound to nickel and zinc affinity columns, whereas the sea urchin protein barely interacted with such columns. Direct determination of metal ions by atomic absorption revealed 2.6 mole zinc/mole protein for human FGFRL1 and 1.7 mole zinc/mole protein for sea urchin FGFRL1. Conclusion The FGFRL1 gene has evolved much earlier than previously assumed. A comparison of the intracellular domain between sea urchin and human FGFRL1 provides interesting insights into the shaping of a novel zinc binding domain. PMID:20021659

  12. Characterization of the Highly Variable Immune Response Gene Family, He185/333, in the Sea Urchin, Heliocidaris erythrogramma

    PubMed Central

    Roth, Mattias O.; Wilkins, Adam G.; Cooke, Georgina M.; Raftos, David A.; Nair, Sham V.

    2014-01-01

    This study characterizes the highly variable He185/333 genes, transcripts and proteins in coelomocytes of the sea urchin, Heliocidaris erythrogramma. Originally discovered in the purple sea urchin, Strongylocentrotus purpuratus, the products of this gene family participate in the anti-pathogen defenses of the host animals. Full-length He185/333 genes and transcripts are identified. Complete open reading frames of He185/333 homologues are analyzed as to their element structure, single nucleotide polymorphisms, indels and sequence repeats and are subjected to diversification analyses. The sequence elements that compose He185/333 are different to those identified for Sp185/333. Differences between Sp185/333 and He185/333 genes are also evident in the complexity of the sequences of the introns. He185/333 proteins show a diverse range of molecular weights on Western blots. The observed sizes and pIs of the proteins differ from predicted values, suggesting post-translational modifications and oligomerization. Immunofluorescence microscopy shows that He185/333 proteins are mainly located on the surface of coelomocyte subpopulations. Our data demonstrate that He185/333 bears the same substantial characteristics as their S. purpuratus homologues. However, we also identify several unique characteristics of He185/333 (such as novel element patterns, sequence repeats, distribution of positively-selected codons and introns), suggesting species-specific adaptations. All sequences in this publication have been submitted to Genbank (accession numbers JQ780171-JQ780321) and are listed in table S1. PMID:25333281

  13. Dynamic evolution of toll-like receptor multigene families in echinoderms.

    PubMed

    Buckley, Katherine M; Rast, Jonathan P

    2012-01-01

    The genome sequence of the purple sea urchin, Strongylocentrotus purpuratus, a large and long-lived invertebrate, provides a new perspective on animal immunity. Analysis of this genome uncovered a highly complex immune system in which the gene families that encode homologs of the pattern recognition receptors that form the core of vertebrate innate immunity are encoded in large multigene families. The sea urchin genome contains 253 Toll-like receptor (TLR) sequences, more than 200 Nod-like receptors and 1095 scavenger receptor cysteine-rich domains, a 10-fold expansion relative to vertebrates. Given their stereotypic protein structure and simple intron-exon architecture, the TLRs are the most tractable of these families for more detailed analysis. A role for these receptors in immune defense is suggested by their similarity to TLRs in other organisms, sequence diversity, and expression in immunologically active tissues, including phagocytes. The complexity of the sea urchin TLR multigene families is largely derived from expansions independent of those in vertebrates and protostomes, although a small family of TLRs with structure similar to that of Drosophila Toll can be traced to an ancient eumetazoan ancestor. Several other echinoderm sequences are now available, including Lytechinus variegatus, as well as partial sequences from two other sea urchin species. Here, we present an analysis of the invertebrate deuterostome TLRs with emphasis on the echinoderms. Representatives of most of the S. purpuratus TLR subfamilies and homologs of the mccTLR sequences are found in L. variegatus, although the L. variegatus TLR gene family is notably smaller (68 TLR sequences). The phylogeny of these genes within sea urchins highlights lineage-specific expansions at higher resolution than is evident at the phylum level. These analyses identify quickly evolving TLR subfamilies that are likely to have novel immune recognition functions and other, more stable, subfamilies that may

  14. Whole-genome positive selection and habitat-driven evolution in a shallow and a deep-sea urchin.

    PubMed

    Oliver, Thomas A; Garfield, David A; Manier, Mollie K; Haygood, Ralph; Wray, Gregory A; Palumbi, Stephen R

    2010-01-01

    Comparisons of genomic sequence between divergent species can provide insight into the action of natural selection across many distinct classes of proteins. Here, we examine the extent of positive selection as a function of tissue-specific and stage-specific gene expression in two closely-related sea urchins, the shallow-water Strongylocentrotus purpuratus and the deep-sea Allocentrotus fragilis, which have diverged greatly in their adult but not larval habitats. Genes that are expressed specifically in adult somatic tissue have significantly higher dN/dS ratios than the genome-wide average, whereas those in larvae are indistinguishable from the genome-wide average. Testis-specific genes have the highest dN/dS values, whereas ovary-specific have the lowest. Branch-site models involving the outgroup S. franciscanus indicate greater selection (ω(FG)) along the A. fragilis branch than along the S. purpuratus branch. The A. fragilis branch also shows a higher proportion of genes under positive selection, including those involved in skeletal development, endocytosis, and sulfur metabolism. Both lineages are approximately equal in enrichment for positive selection of genes involved in immunity, development, and cell-cell communication. The branch-site models further suggest that adult-specific genes have experienced greater positive selection than those expressed in larvae and that ovary-specific genes are more conserved (i.e., experienced greater negative selection) than those expressed specifically in adult somatic tissues and testis. Our results chart the patterns of protein change that have occurred after habitat divergence in these two species and show that the developmental or functional context in which a gene acts can play an important role in how divergent species adapt to new environments. PMID:20935062

  15. El Ventorrillo, a paleostructure of Popocatépetl volcano: insights from geochronology and geochemistry

    NASA Astrophysics Data System (ADS)

    Sosa-Ceballos, G.; Macías, J. L.; García-Tenorio, F.; Layer, P.; Schaaf, P.; Solís-Pichardo, G.; Arce, J. L.

    2015-10-01

    Volcán Popocatépetl (México) was constructed over the remains of a volcanic paleostructure. Based on fieldwork, 40Ar/39Ar dating, U-Pb dating, and geochemistry, we have determined the age, chemistry, and location of this paleostructure and named it El Ventorrillo. Most remnants of El Ventorrillo are covered by deposits from subsequent activity of Popocatépetl, except for the El Abanico scarp and the Barranca de Nexpayantla, where the stratigraphy of El Ventorrillo eruptive products can be investigated. Inception of volcanism at El Ventorrillo occurred at 331 ± 10 ka with emission of the Nexpayantla andesitic lavas, and continued with extrusion of the Yoloxochitl (267 ± 31 ka) and microwave (227 ± 6 ka) domes. Intrusion of dikes occurred at 298 ± 94 and 230 ± 3 ka. Activity at El Ventorrillo continued with the emission of lavas that built the El Abanico scarp (193 ± 29 to 96 ± 8 ka) and continued until the Tutti Frutti eruption destroyed the cone 14.1 kyr ago. El Ventorrillo magmas produced rocks divided into two mineralogical groups. The first group contains biotite-amphibole-rich rocks and the second group consists of biotite-amphibole-free lavas. The rocks that contain biotite and amphibole are older than 198 ± 13 ka, whereas the rocks with no hydrous phases are younger than 227 ± 6 ka and contain skarn and granodiorite xenoliths. We interpret the change to an anhydrous mineral assemblage and the occurrence of skarn and granodiorite xenoliths as evidence for the formation of a new, shallower reservoir. A granodiorite xenolith was chosen for 40Ar/39Ar dating and U-Pb zircon analyses. The U-Pb method yielded an age of 540 ± 110 ka and the 40Ar/39Ar an age of 109 ± 24 ka. These ages are interpreted to indicate granodiorite crystallization (540 ± 110 ka), which metamorphosed the calcareous basement beneath Popocatépetl into skarn and an influx of magma (109 ± 24 ka) that reheated the granodiorite. Major and trace elements, Sr, Nd, and Pb isotopic

  16. Resistance to Genotoxic Stresses in Arctica islandica, the Longest Living Noncolonial Animal: Is Extreme Longevity Associated With a Multistress Resistance Phenotype?

    PubMed Central

    Ungvari, Zoltan; Sosnowska, Danuta; Mason, Jeffrey B.; Gruber, Heike

    2013-01-01

    Bivalve molluscs are newly discovered models of successful aging. Here, we test the hypothesis that extremely long-lived bivalves are not uniquely resistant to oxidative stressors (eg, tert-butyl hydroperoxide, as demonstrated in previous studies) but exhibit a multistress resistance phenotype. We contrasted resistance (in terms of organismal mortality) to genotoxic stresses (including topoisomerase inhibitors, agents that cross-link DNA or impair genomic integrity through DNA alkylation or methylation) and to mitochondrial oxidative stressors in three bivalve mollusc species with dramatically differing life spans: Arctica islandica (ocean quahog), Mercenaria mercenaria (northern quahog), and the Atlantic bay scallop, Argopecten irradians irradians (maximum species life spans: >500, >100, and ~2 years, respectively). With all stressors, the short-lived A i irradians were significantly less resistant than the two longer lived species. Arctica islandica were consistently more resistant than M mercenaria to mortality induced by oxidative stressors as well as DNA methylating agent nitrogen mustard and the DNA alkylating agent methyl methanesulfonate. The same trend was not observed for genotoxic agents that act through cross-linking DNA. In contrast, M mercenaria tended to be more resistant to epirubicin and genotoxic stressors, which cause DNA damage by inhibiting topoisomerases. To our knowledge, this is the first study comparing resistance to genotoxic stressors in bivalve mollusc species with disparate longevities. In line with previous studies of comparative stress resistance and longevity, our data extends, at least in part, the evidence for the hypothesis that an association exists between longevity and a general resistance to multiplex stressors, not solely oxidative stress. This work also provides justification for further investigation into the interspecies differences in stress response signatures induced by a diverse array of stressors in short-lived and

  17. De Novo Assembly and Characterization of Two Transcriptomes Reveal Multiple Light-Mediated Functions in the Scallop Eye (Bivalvia: Pectinidae)

    PubMed Central

    Pairett, Autum N.; Serb, Jeanne M.

    2013-01-01

    Background The eye has evolved across 13 separate lineages of molluscs. Yet, there have been very few studies examining the molecular machinary underlying eye function of this group, which is due, in part, to a lack of genomic resources. The scallop (Bivalvia: Pectinidae) represents a compeling molluscan model to study photoreception due to its morphologically novel and separately evolved mirror-type eye. We sequenced the adult eye transcriptome of two scallop species to: 1) identify the phototransduction pathway components; 2) identify any additional light detection functions; and 3) test the hypothesis that molluscs possess genes not found in other animal lineages. Results A total of 3,039 contigs from the bay scallop, Argopecten irradians and 26,395 contigs from the sea scallop, Placopecten magellanicus were produced by 454 sequencing. Targeted BLAST searches and functional annotation using Gene Ontology (GO) terms and KEGG pathways identified transcripts from three light detection systems: two phototransduction pathways and the circadian clock, a previously unrecognized function of the scallop eye. By comparing the scallop transcriptomes to molluscan and non-molluscan genomes, we discovered that a large proportion of the transcripts (7,776 sequences) may be specific to the scallop lineage. Nearly one-third of these contain transmembrane protein domains, suggesting these unannotated transcripts may be sensory receptors. Conclusions Our data provide the most comprehensive transcriptomic resource currently available from a single molluscan eye type. Candidate genes potentially involved in sensory reception were identified, and are worthy of further investigation. This resource, combined with recent phylogenetic and genomic data, provides a strong foundation for future investigations of the function and evolution of molluscan photosensory systems in this morphologically and taxonomically diverse phylum. PMID:23922823

  18. Comparative effects of the toxic dinoflagellate Karenia brevis on clearance rates in juveniles of four bivalve molluscs from Florida, USA.

    PubMed

    Leverone, Jay R; Shumway, Sandra E; Blake, Norman J

    2007-04-01

    The effects of Karenia brevis (Gymnodiniales, Gymnodiniaceae) on the feeding activity of juveniles of four species of bivalve mollusc were examined in the laboratory to assess the potential impacts on these important shellfish populations from Florida. Clearance rates were determined under short-term (one hour) static and long-term (two days) flow-through conditions using both whole and lysed cultures of K. brevis. Under short-term conditions, the bay scallop, Argopecten irradians, was the most sensitive species, exhibiting a 79% reduction in clearance rate at 1000 cells ml(-1) of whole K. brevis culture compared to the control (no K. brevis). The eastern oyster, Crassostrea virginica, was the least responsive, showing a 38% reduction in clearance rate between the same treatments. The green mussel, Perna viridis, and the northern quahog, Mercenaria mercenaria, displayed intermediate responses. Similar results were also observed during long-term exposures to a continuous supply of K. brevis. Bay scallops showed a significant decline in clearance rate at 100 cells ml(-1) after 24h exposure; clearance rate of oysters was not affected by K. brevis at this concentration. No mortality was observed for any species during these brief exposures. The prospect for recovery of bay scallop populations in Florida estuaries where they were once abundant may be hampered by recurring blooms of K. brevis. Reduced clearance rates in M. mercenaria at high K. brevis densities could translate into poor growth of cultured Florida hard clams. On the other hand, P. viridis, which also showed reduced clearance rates at high K. brevis concentrations, might be negatively impacted by K. brevis blooms, thereby affecting their ability to spread into estuaries hampered by recurring toxic algal blooms. PMID:17182073

  19. Physiological response and resilience of early life-stage Eastern oysters (Crassostrea virginica) to past, present and future ocean acidification

    PubMed Central

    Gobler, Christopher J.; Talmage, Stephanie C.

    2014-01-01

    The Eastern oyster, Crassostrea virginica (Gmelin, 1791), is the second most valuable bivalve fishery in the USA and is sensitive to high levels of partial pressure of CO2 (pCO2). Here we present experiments that comprehensively examined how the ocean's past, present and projected (21st and 22nd centuries) CO2 concentrations impact the growth and physiology of larval stages of C. virginica. Crassostrea virginica larvae grown in present-day pCO2 concentrations (380 μatm) displayed higher growth and survival than individuals grown at both lower (250 μatm) and higher pCO2 levels (750 and 1500 μatm). Crassostrea virginica larvae manifested calcification rates, sizes, shell thicknesses, metamorphosis, RNA:DNA ratios and lipid contents that paralleled trends in survival, with maximal values for larvae grown at 380 μatm pCO2 and reduced performance in higher and lower pCO2 levels. While some physiological differences among oysters could be attributed to CO2-induced changes in size or calcification rates, the RNA:DNA ratios at ambient pCO2 levels were elevated, independent of these factors. Likewise, the lipid contents of individuals exposed to high pCO2 levels were depressed even when differences in calcification rates were considered. These findings reveal the cascading, interdependent impact that high CO2 can have on oyster physiology. Crassostrea virginica larvae are significantly more resistant to elevated pCO2 than other North Atlantic bivalves, such as Mercenaria mercenaria and Argopecten irradians, a finding that may be related to the biogeography and/or evolutionary history of these species and may have important implications for future bivalve restoration and aquaculture efforts. PMID:27293625

  20. Hypoxia and Acidification Have Additive and Synergistic Negative Effects on the Growth, Survival, and Metamorphosis of Early Life Stage Bivalves

    PubMed Central

    Gobler, Christopher J.; DePasquale, Elizabeth L.; Griffith, Andrew W.; Baumann, Hannes

    2014-01-01

    Low oxygen zones in coastal and open ocean ecosystems have expanded in recent decades, a trend that will accelerate with climatic warming. There is growing recognition that low oxygen regions of the ocean are also acidified, a condition that will intensify with rising levels of atmospheric CO2. Presently, however, the concurrent effects of low oxygen and acidification on marine organisms are largely unknown, as most prior studies of marine hypoxia have not considered pH levels. We experimentally assessed the consequences of hypoxic and acidified water for early life stage bivalves (bay scallops, Argopecten irradians, and hard clams, Mercenaria mercenaria), marine organisms of significant economic and ecological value and sensitive to climate change. In larval scallops, experimental and naturally-occurring acidification (pH, total scale  = 7.4–7.6) reduced survivorship (by >50%), low oxygen (30–50 µM) inhibited growth and metamorphosis (by >50%), and the two stressors combined produced additively negative outcomes. In early life stage clams, however, hypoxic waters led to 30% higher mortality, while acidified waters significantly reduced growth (by 60%). Later stage clams were resistant to hypoxia or acidification separately but experienced significantly (40%) reduced growth rates when exposed to both conditions simultaneously. Collectively, these findings demonstrate that the consequences of low oxygen and acidification for early life stage bivalves, and likely other marine organisms, are more severe than would be predicted by either individual stressor and thus must be considered together when assessing how ocean animals respond to these conditions both today and under future climate change scenarios. PMID:24416169

  1. Effects of pH and bicarbonate on mitochondrial functions of marine bivalves.

    PubMed

    Haider, Fouzia; Falfushynska, Halina; Ivanina, Anna V; Sokolova, Inna M

    2016-08-01

    Estuarine organisms including mollusks are exposed to periodic oxygen deficiency (hypoxia) that leads to a decrease in intracellular pH and accumulation of bicarbonate (HCO3(-)). These changes can affect cellular bioenergetics; however, their effects on mitochondria of estuarine mollusks are not well understood. We determined the interactive effects of bicarbonate (0-10mM) and pH (7.2 and 6.5) on mitochondrial oxygen consumption (ṀO2), membrane potential (Δψ) and production of reactive oxygen species (ROS) in two common estuarine bivalves - hard clams Mercenaria mercenaria, and bay scallops Argopecten irradians. In both species, elevated HCO3(-) levels suppressed ADP-stimulated (state 3) ṀO2 but had little effect on the resting (state 4) respiration. These effects were not mediated by the soluble adenylyl cyclase or cyclic AMP. Effects of the low pH (6.5) on mitochondrial traits were species-specific and depended on the substrate oxidized by the mitochondria. Mild acidosis (pH6.5) had minimal effects on ṀO2 and Δψ of the bivalve mitochondria oxidizing pyruvate but led to increased rates of ROS production in clams (ROS production could not be measured in scallops). In succinate-respiring mitochondria of clams, mild acidosis suppressed ṀO2 and increased mitochondrial coupling, while in scallop mitochondria the effects of low pH were opposite. Suppression of mitochondrial oxidative phosphorylation by bicarbonate and/or acidosis may contribute to the metabolic rate depression during shell closure or environmental hypoxia/hypercapnia. These findings have implications for understanding the physiological mechanisms involved in regulation of mitochondrial bioenergetics during hypoxia exposure in estuarine bivalves. PMID:27044911

  2. Cladistic analysis and description of three new species of the Chilean genus Nanophareus (Opiliones: Gonyleptidae: Pachylinae).

    PubMed

    Hara, Marcos Ryotaro

    2016-01-01

    Three new species of the Chilean Pachylinae genus, Nanophareus Roewer, 1929 are described: N. bicornutus sp. nov. (Valle de Aconcagua, Zapallar, V Región de Valparaíso), N. maipu sp. nov. (La Rinconada, Quebrada de la Plata, Maipu, Región Metropolitana-Santiago), and N. polyhastatus sp. nov. (El Abanico, VIII Región de Bio-Bío). These three new species were included in a cladistic analysis that resulted in two equally most parsimonious trees (238 steps, C.I. = 0.38; R.I. = 0.51), corroborating the monophyly of Nanophareus. The proposed synapomorphies for Nanophareus remain largely unchanged: an external row of enlarged tubercles inserted amongst small ones on the lateral margin of dorsal scutum; the ventro-basal margin of the pedipalpal tibia curved at 90° in lateral view; and retrolateral seta on pedipalpal tibia with an apically bifid socket (socket and seta longer than pedipalpal tibia length), with additional small setae distally. The sister group of Nanophareus, as well as its subfamilial placement, are still unsettled issues that are here further discussed. PMID:27394767

  3. Ultrastructural localization of intracellular calcium stores by a new cytochemical method.

    PubMed

    Poenie, M; Epel, D

    1987-09-01

    We describe a new cytochemical method for ultrastructural localization of intracellular calcium stores. This method uses fluoride ions for in situ precipitation of intracellular calcium during fixation. Comparisons made using oxalate, antimonate, or fluoride showed that fluoride was clearly superior for intracellular calcium localization in eggs of the sea urchin Strongylocentrotus purpuratus. Whereas oxalate generally gave no intracellular precipitate and antimonate gave copious but random precipitate, three prominent calcium stores were detected using fluoride: the tubular endoplasmic reticulum, the cortical granules, and large, clear, acidic vesicles of unknown function. The mitochondria of these eggs generally showed no detectable calcium deposits. X-ray spectra confirmed the presence of calcium in the fluoride precipitates, although in some cases magnesium was also detected. Rat skeletal muscle and sea urchin sperm were used to test the reliability of the fluoride method for calcium localization. In rat skeletal muscle, most fluoride precipitate was confined to the sarcoplasmic reticulum. Using sea urchin sperm, which transport calcium into the mitochondria after exposure to egg jelly to induce the acrosome reaction, the expected result was also obtained. Before the acrosome reaction, sperm mitochondria contain no detectable calcium-containing precipitate. Within 4 min after induction of the acrosome reaction, the expected result was also obtained. Before the acrosome reaction, sperm mitochondria displayed many foci of calcium-containing precipitate. The use of fluoride for intracellular calcium localization therefore appears to be a substantial improvement over previous cytochemical methods. PMID:3611737

  4. Status and applications of echinoid (phylum echinodermata) toxicity test methods

    SciTech Connect

    Bay, S.; Burgess, R.; Nacci, D.

    1993-01-01

    The use of echinoderms for toxicity testing has focused primarily on sea urchins and sand dollars (Strongylocentrotus purpuratus, Arbacia punctulata, Lytechinus pictus, and Dendraster excentricus, for example). The status and relative sensitivity of various test methods are described. The most frequently used test methods consist of short-term exposures of sea urchin sperm or embryos; these tests can be easily conducted at all times of the year by using species with complementary spawning cycles or laboratory conditioned populations of a single species. Data from reference toxicant and effluent toxicity tests are summarized. Information on the precision and sensitivity of echinoid test methods are limited and preclude rigorous comparisons with other test methods. The available data indicate that the sensitivity and precision of these methods are comparable to short-term chronic methods for other marine invertebrates and fish. Recent application of the sperm test in toxicity identification evaluations (TIEs) and studies of effluent toxicity decay and sediment toxicity illustrate the versatility of this rapid (10 to 60 min exposure) test method. Embryo tests typically use a 48 to 96 h exposure period and measure the occurrence of embryo malformations. Most recent applications of the embryo test have been for the assessment of sediment elutriate toxicity. Adult echinoderms are not frequently used to assess effluent or receiving water toxicity. Recent studies have had success in using the adult life stage of urchins and sand dollars to assess the effects of contaminated sediment on growth, behavior, and bioaccumulation.

  5. Mechanism of Calcite Co-Orientation in the Sea Urchin Tooth

    SciTech Connect

    Killian, Christopher; Metzler, Rebecca; Gong, Y. U. T.; Olson, Ian; Aizenberg, Joanna; Politi, Yael; Wilt, Fred; Scholl, Andreas; Young, Anthony; Doran, Andrew; Kunz, Martin; Tamura, Nobumichi; Coppersmith, Susan; Gilbert, P. U. P. A.

    2009-12-01

    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{sub 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 ({mu}XRD). 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{sub 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.

  6. Evolutionary rewiring of gene regulatory network linkages at divergence of the echinoid subclasses

    PubMed Central

    Erkenbrack, Eric M.; Davidson, Eric H.

    2015-01-01

    Evolution of animal body plans occurs with changes in the encoded genomic programs that direct development, by alterations in the structure of encoded developmental gene-regulatory networks (GRNs). However, study of this most fundamental of evolutionary processes requires experimentally tractable, phylogenetically divergent organisms that differ morphologically while belonging to the same monophyletic clade, plus knowledge of the relevant GRNs operating in at least one of the species. These conditions are met in the divergent embryogenesis of the two extant, morphologically distinct, echinoid (sea urchin) subclasses, Euechinoidea and Cidaroidea, which diverged from a common late Paleozoic ancestor. Here we focus on striking differences in the mode of embryonic skeletogenesis in a euechinoid, the well-known model Strongylocentrotus purpuratus (Sp), vs. the cidaroid Eucidaris tribuloides (Et). At the level of descriptive embryology, skeletogenesis in Sp and Et has long been known to occur by distinct means. The complete GRN controlling this process is known for Sp. We carried out targeted functional analyses on Et skeletogenesis to identify the presence, or demonstrate the absence, of specific regulatory linkages and subcircuits key to the operation of the Sp skeletogenic GRN. Remarkably, most of the canonical design features of the Sp skeletogenic GRN that we examined are either missing or operate differently in Et. This work directly implies a dramatic reorganization of genomic regulatory circuitry concomitant with the divergence of the euechinoids, which began before the end-Permian extinction. PMID:26170318

  7. Coastal Upwelling Drives Intertidal Assemblage Structure and Trophic Ecology

    PubMed Central

    Reddin, Carl J.; Docmac, Felipe; O’Connor, Nessa E.; Bothwell, John H.; Harrod, Chris

    2015-01-01

    Similar environmental driving forces can produce similarity among geographically distant ecosystems. Coastal oceanic upwelling, for example, has been associated with elevated biomass and abundance patterns of certain functional groups, e.g., corticated macroalgae. In the upwelling system of Northern Chile, we examined measures of intertidal macrobenthic composition, structure and trophic ecology across eighteen shores varying in their proximity to two coastal upwelling centres, in a hierarchical sampling design (spatial scales of >1 and >10 km). The influence of coastal upwelling on intertidal communities was confirmed by the stable isotope values (δ13C and δ15N) of consumers, including a dominant suspension feeder, grazers, and their putative resources of POM, epilithic biofilm, and macroalgae. We highlight the utility of muscle δ15N from the suspension feeding mussel, Perumytilus purpuratus, as a proxy for upwelling, supported by satellite data and previous studies. Where possible, we used corrections for broader-scale trends, spatial autocorrelation, ontogenetic dietary shifts and spatial baseline isotopic variation prior to analysis. Our results showed macroalgal assemblage composition, and benthic consumer assemblage structure, varied significantly with the intertidal influence of coastal upwelling, especially contrasting bays and coastal headlands. Coastal topography also separated differences in consumer resource use. This suggested that coastal upwelling, itself driven by coastline topography, influences intertidal communities by advecting nearshore phytoplankton populations offshore and cooling coastal water temperatures. We recommend the isotopic values of benthic organisms, specifically long-lived suspension feeders, as in situ alternatives to offshore measurements of upwelling influence. PMID:26214806

  8. Speract, a sea urchin egg peptide that regulates sperm motility, also stimulates sperm mitochondrial metabolism.

    PubMed

    García-Rincón, Juan; Darszon, Alberto; Beltrán, Carmen

    2016-04-01

    Sea urchin sperm have only one mitochondrion, that in addition to being the main source of energy, may modulate intracellular Ca(2+) concentration ([Ca(2+)]i) to regulate their motility and possibly the acrosome reaction. Speract is a decapeptide from the outer jelly layer of the Strongylocentrotus purpuratus egg that upon binding to its receptor in the sperm, stimulates sperm motility, respiration and ion fluxes, among other physiological events. Altering the sea urchin sperm mitochondrial function with specific inhibitors of this organelle, increases [Ca(2+)]i in an external Ca(2+) concentration ([Ca(2+)]ext)-dependent manner (Ardón, et al., 2009. BBActa 1787: 15), suggesting that the mitochondrion is involved in sperm [Ca(2+)]i homeostasis. To further understand the interrelationship between the mitochondrion and the speract responses, we measured mitochondrial membrane potential (ΔΨ) and NADH levels. We found that the stimulation of sperm with speract depolarizes the mitochondrion and increases the levels of NADH. Surprisingly, these responses are independent of external Ca(2+) and are due to the increase in intracellular pH (pHi) induced by speract. Our findings indicate that speract, by regulating pHi, in addition to [Ca(2+)]i, may finely modulate mitochondrial metabolism to control motility and ensure that sperm reach the egg and fertilize it. PMID:26772728

  9. Analysis of Cytoskeletal and Motility Proteins in the Sea Urchin Genome Assembly

    PubMed Central

    RL, Morris; MP, Hoffman; RA, Obar; SS, McCafferty; IR, Gibbons; AD, Leone; J, Cool; EL, Allgood; AM, Musante; KM, Judkins; BJ, Rossetti; AP, Rawson; DR, Burgess

    2007-01-01

    The sea urchin embryo is a classical model system for studying the role of the cytoskeleton in such events as fertilization, mitosis, cleavage, cell migration and gastrulation. We have conducted an analysis of gene models derived from the Strongylocentrotus purpuratus genome assembly and have gathered strong evidence for the existence of multiple gene families encoding cytoskeletal proteins and their regulators in sea urchin. While many cytoskeletal genes have been cloned from sea urchin with sequences already existing in public databases, genome analysis reveals a significantly higher degree of diversity within certain gene families. Furthermore, genes are described corresponding to homologs of cytoskeletal proteins not previously documented in sea urchins. To illustrate the varying degree of sequence diversity that exists within cytoskeletal gene families, we conducted an analysis of genes encoding actins, specific actin-binding proteins, myosins, tubulins, kinesins, dyneins, specific microtubule-associated proteins, and intermediate filaments. We conducted ontological analysis of select genes to better understand the relatedness of urchin cytoskeletal genes to those of other deuterostomes. We analyzed developmental expression (EST) data to confirm the existence of select gene models and to understand their differential expression during various stages of early development. PMID:17027957

  10. Multidrug Efflux Transporters Limit Accumulation of Inorganic, but Not Organic, Mercury in Sea Urchin Embryos

    PubMed Central

    Bošnjak, Ivana; Uhlinger, Kevin R.; Heim, Wesley; Smital, Tvrtko; Franekić-Čolić, Jasna; Coale, Kenneth; Epel, David; Hamdoun, Amro

    2011-01-01

    Mercuric compounds are persistent global pollutants that accumulate in marine organisms and in humans who consume them. While the chemical cycles and speciation of mercury in the oceans are relatively well described, the cellular mechanisms that govern which forms of mercury accumulate in cells and why they persist are less understood. In this study we examined the role of multidrug efflux transport in the differential accumulation of inorganic (HgCl2) and organic (CH3HgCl) mercury in sea urchin (Strongylocentrotus purpuratus) embryos. We found that inhibition of MRP/ABCC-type transporters increases intracellular accumulation of inorganic mercury but had no effect on accumulation of organic mercury. Similarly, pharmacological inhibition of metal conjugating enzymes by ligands GST/GSH significantly increases this antimitotic potency of inorganic mercury, but had no effect on the potency of organic mercury. Our results point to MRP-mediated elimination of inorganic mercury conjugates as a cellular basis for differences in the accumulation and potency of the two major forms of mercury found in marine environments. PMID:19924972

  11. High-Density Genetic Mapping with Interspecific Hybrids of Two Sea Urchins, Strongylocentrotus nudus and S. intermedius, by RAD Sequencing.

    PubMed

    Zhou, Zunchun; Liu, Shikai; Dong, Ying; Gao, Shan; Chen, Zhong; Jiang, Jingwei; Yang, Aifu; Sun, Hongjuan; Guan, Xiaoyan; Jiang, Bei; Wang, Bai

    2015-01-01

    Sea urchins have long been used as research model organisms for developmental biology and evolutionary studies. Some of them are also important aquaculture species in East Asia. In this work, we report the construction of RAD-tag based high-density genetic maps by genotyping F1 interspecific hybrids derived from a crossing between a female sea urchin Strongylocentrotus nudus and a male Strongylocentrotus intermedius. With polymorphisms present in these two wild individuals, we constructed a female meiotic map containing 3,080 markers for S. nudus, and a male meiotic map for S. intermedius which contains 1,577 markers. Using the linkage maps, we were able to anchor a total of 1,591 scaffolds (495.9 Mb) accounting for 60.8% of the genome assembly of Strongylocentrotus purpuratus. A genome-wide scan resulted in the identification of one putative QTL for body size which spanned from 25.3 cM to 30.3 cM. This study showed the efficiency of RAD-Seq based high-density genetic map construction using F1 progenies for species with no prior genomic information. The genetic maps are essential for QTL mapping and are useful as framework to order and orientate contiguous scaffolds from sea urchin genome assembly. The integration of the genetic map with genome assembly would provide an unprecedented opportunity to conduct QTL analysis, comparative genomics, and population genetics studies. PMID:26398139

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

    PubMed Central

    Wilson, David R.; Norton, Darrell D.; Fugmann, Sebastian D.

    2008-01-01

    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. PMID:18499250

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

    PubMed

    Wilson, David R; Norton, Darrell D; Fugmann, Sebastian D

    2008-01-01

    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. PMID:18499250

  14. The Insect Chemoreceptor Superfamily Is Ancient in Animals.

    PubMed

    Robertson, Hugh M

    2015-11-01

    The insect chemoreceptor superfamily consists of 2 gene families, the highly diverse gustatory receptors (GRs) found in all arthropods with sequenced genomes and the odorant receptors that evolved from a GR lineage and have been found only in insects to date. Here, I describe relatives of the insect chemoreceptor superfamily, specifically the basal GR family, in diverse other animals, showing that the superfamily dates back at least to early animal evolution. GR-Like (GRL) genes are present in the genomes of the placozoan Trichoplax adhaerens, an anemone Nematostella vectensis, a coral Acropora digitifera, a polychaete Capitella teleta, a leech Helobdella robusta, the nematode Caenorhabditis elegans (and many other nematodes), 3 molluscs (a limpet Lottia gigantea, an oyster Crassostrea gigas, and the sea hare Aplysia californica), the sea urchin Strongylocentrotus purpuratus, and the sea acorn Saccoglossus kowalevskii. While some of these animals contain multiple divergent GRL lineages, GRLs have been lost entirely from other animal lineages such as vertebrates. GRLs are absent from the ctenophore Mnemiopsis leidyi, the demosponge Amphimedon queenslandica, and 2 available chaonoflagellate genomes, so it remains unclear whether this superfamily originated before or during animal evolution. PMID:26354932

  15. Lesions of Copper Toxicosis in Captive Marine Invertebrates With Comparisons to Normal Histology.

    PubMed

    LaDouceur, E E B; Wynne, J; Garner, M M; Nyaoke, A; Keel, M K

    2016-05-01

    Despite increasing concern for coral reef ecosystem health within the last decade, there is scant literature concerning the histopathology of diseases affecting the major constituents of coral reef ecosystems, particularly marine invertebrates. This study describes histologic findings in 6 species of marine invertebrates (California sea hare [Aplysia californica], purple sea urchin [Strongylocentrotus purpuratus], sunburst anemone [Anthopleura sola], knobby star [Pisaster giganteus], bat star [Asterina miniata], and brittle star [Ophiopteris papillosa]) with spontaneous copper toxicosis, 4 purple sea urchins with experimentally induced copper toxicosis, and 1 unexposed control of each species listed. The primary lesions in the California sea hare with copper toxicosis were branchial and nephridial necrosis. Affected echinoderms shared several histologic lesions, including epidermal necrosis and ulceration and increased numbers of coelomocytes within the water-vascular system. The sunburst anemone with copper toxicosis had necrosis of both epidermis and gastrodermis, as well as expulsion of zooxanthellae from the gastrodermis. In addition to the lesions attributed to copper toxicosis, our results describe normal microscopic features of these animals that may be useful for histopathologic assessment of marine invertebrates. PMID:26459519

  16. The carboxylesterase/cholinesterase gene family in invertebrate deuterostomes.

    PubMed

    Johnson, Glynis; Moore, Samuel W

    2012-06-01

    Carboxylesterase/cholinesterase family members are responsible for controlling the nerve impulse, detoxification and various developmental functions, and are a major target of pesticides and chemical warfare agents. Comparative structural analysis of these enzymes is thus important. The invertebrate deuterostomes (phyla Echinodermata and Hemichordata and subphyla Urochordata and Cephalochordata) lie in the transition zone between invertebrates and vertebrates, and are thus of interest to the study of evolution. Here we have investigated the carboxylesterase/cholinesterase gene family in the sequenced genomes of Strongylocentrotus purpuratus (Echinodermata), Saccoglossus kowalevskii (Hemichordata), Ciona intestinalis (Urochordata) and Branchiostoma floridae (Cephalochordata), using sequence analysis of the catalytic apparatus and oligomerisation domains, and phylogenetic analysis. All four genomes show blurring of structural boundaries between cholinesterases and carboxylesterases, with many intermediate enzymes. Non-enzymatic proteins are well represented. The Saccoglossus and Branchiostoma genomes show evidence of extensive gene duplication and retention. There is also evidence of domain shuffling, resulting in multidomain proteins consisting either of multiple carboxylesterase domains, or of carboxylesterase/cholinesterase domains linked to other domains, including RING finger, chitin-binding, immunoglobulin, fibronectin type 3, CUB, cysteine-rich-Frizzled, caspase activation and 7tm-1, amongst others. Such gene duplication and domain shuffling in the carboxylesterase/cholinesterase family appears to be unique to the invertebrate deuterostomes, and we hypothesise that these factors may have contributed to the evolution of the morphological complexity, particularly of the nervous system and neural crest, of the vertebrates. PMID:22210164

  17. Discovery of sea urchin NGFFFamide receptor unites a bilaterian neuropeptide family

    PubMed Central

    Semmens, Dean C.; Beets, Isabel; Rowe, Matthew L.; Blowes, Liisa M.; Oliveri, Paola; Elphick, Maurice R.

    2015-01-01

    Neuropeptides are ancient regulators of physiology and behaviour, but reconstruction of neuropeptide evolution is often difficult owing to lack of sequence conservation. Here, we report that the receptor for the neuropeptide NGFFFamide in the sea urchin Strongylocentrotus purpuratus (phylum Echinodermata) is an orthologue of vertebrate neuropeptide-S (NPS) receptors and crustacean cardioactive peptide (CCAP) receptors. Importantly, this has facilitated reconstruction of the evolution of two bilaterian neuropeptide signalling systems. Genes encoding the precursor of a vasopressin/oxytocin-type neuropeptide and its receptor duplicated in a common ancestor of the Bilateria. One copy of the precursor retained ancestral features, as seen in highly conserved vasopressin/oxytocin–neurophysin-type precursors. The other copy diverged, but this took different courses in protostomes and deuterostomes. In protostomes, the occurrence of a disulfide bridge in neuropeptide product(s) of the precursor was retained, as in CCAP, but with loss of the neurophysin domain. In deuterostomes, we see the opposite scenario—the neuropeptides lost the disulfide bridge, and neurophysin was retained (as in the NGFFFamide precursor) but was subsequently lost in vertebrate NPS precursors. Thus, the sea urchin NGFFFamide precursor and receptor are ‘missing links’ in the evolutionary history of neuropeptides that control ecdysis in arthropods (CCAP) and regulate anxiety in humans (NPS). PMID:25904544

  18. Unusual Gene Order and Organization of the Sea Urchin HoxCluster

    SciTech Connect

    Richardson, Paul M.; Lucas, Susan; Cameron, R. Andrew; Rowen,Lee; Nesbitt, Ryan; Bloom, Scott; Rast, Jonathan P.; Berney, Kevin; Arenas-Mena, Cesar; Martinez, Pedro; Davidson, Eric H.; Peterson, KevinJ.; Hood, Leroy

    2005-05-10

    The highly consistent gene order and axial colinear expression patterns found in vertebrate hox gene clusters are less well conserved across the rest of bilaterians. We report the first deuterostome instance of an intact hox cluster with a unique gene order where the paralog groups are not expressed in a sequential manner. The finished sequence from BAC clones from the genome of the sea urchin, Strongylocentrotus purpuratus, reveals a gene order wherein the anterior genes (Hox1, Hox2 and Hox3) lie nearest the posterior genes in the cluster such that the most 3' gene is Hox5. (The gene order is : 5'-Hox1,2, 3, 11/13c, 11/13b, '11/13a, 9/10, 8, 7, 6, 5 - 3)'. The finished sequence result is corroborated by restriction mapping evidence and BAC-end scaffold analyses. Comparisons with a putative ancestral deuterostome Hox gene cluster suggest that the rearrangements leading to the sea urchin gene order were many and complex.

  19. Unusual Gene Order and Organization of the Sea Urchin Hox Cluster

    SciTech Connect

    Cameron, R A; Rowen, L; Nesbitt, R; Bloom, S; Rast, J P; Berney, K; Arenas-Mena, C; Martinez, P; Lucas, S; Richardson, P M; Davidson, E H; Peterson, K J; Hood, L

    2005-10-11

    The highly consistent gene order and axial colinear expression patterns found in vertebrate hox gene clusters are less well conserved across the rest of bilaterians. We report the first deuterostome instance of an intact hox cluster with a unique gene order where the paralog groups are not expressed in a sequential manner. The finished sequence from BAC clones from the genome of the sea urchin, Strongylocentrotus purpuratus, reveals a gene order wherein the anterior genes (Hox1, Hox2 and Hox3) lie nearest the posterior genes in the cluster such that the most 3 gene is Hox5. (The gene order is : 5-Hox1, 2, 3, 11/13c, 11/13b, 11/13a, 9/10, 8, 7, 6, 5 - 3). The finished sequence result is corroborated by restriction mapping evidence and BAC-end scaffold analyses. Comparisons with a putative ancestral deuterostome Hox gene cluster suggest that the rearrangements leading to the sea urchin gene order were many and complex.

  20. A Method for Microinjection of Patiria minata Zygotes

    PubMed Central

    Cheatle Jarvela, Alys M.; Hinman, Veronica

    2014-01-01

    Echinoderms have long been a favorite model system for studies of reproduction and development, and more recently for the study of gene regulation and evolution of developmental processes. The sea star, Patiria miniata, is gaining prevalence as a model system for these types of studies which were previously performed almost exclusively in the sea urchins, Strongylocentrotus purpuratus and Lytechinus variegatus. An advantage of these model systems is the ease of producing modified embryos in which a particular gene is up or downregulated, labeling a group of cells, or introducing a reporter gene. A single microinjection method is capable of creating a wide variety of such modified embryos. Here, we present a method for obtaining gametes from P. miniata, producing zygotes, and introducing perturbing reagents via microinjection. Healthy morphant embryos are subsequently isolated for quantitative and qualitative studies of gene function. The availability of genome and transcriptome data for this organism has increased the types of studies that are performed and the ease of executing them. PMID:25226153

  1. Preservation and visualization of the sea urchin embryo blastocoelic extracellular matrix.

    PubMed

    Cherr, G N; Summers, R G; Baldwin, J D; Morrill, J B

    1992-06-15

    Several methods were utilized to visualize the structure and orientation of the blastocoelic extracellular matrix (ECM) in Strongylocentrotus purpuratus embryos at the mesenchyme blastula stage. Rapid freezing in liquid propane cooled to LN2 temperatures followed by freeze substitution was used to preserve the ECM without shrinkage due to dehydration. Scanning, transmission, and light microscopy were employed to elucidate the ECMs' structure. The blastocoelic ECM consisted of parallel fibrillar sheets that were interconnected by finer filaments and oriented along the animal-vegetal axis. The ECM completely filled the blastocoelic cavity as viewed by scanning electron microscopy. The basal lamina could be distinguished from the blastocoelic ECM as a thin coat on the plasma membrane of epithelial cells; the ECM was in contact with this coat. In contrast, the blastocoelic ECM attached directly to the plasma membrane of primary mesenchyme cells (PMC) which did not possess a basal lamina. The blastocoelic ECM was isolated as an intact "bag" and probed in a hydrated state with Con A and alcian blue. Confocal microscopy confirmed that the entire blastocoel was filled with a fibrillar ECM. These approaches offer advantages for future studies of the ECMs of sea urchin embryos and their roles in gastrulation. PMID:1617206

  2. Transcriptome analysis of the scleractinian coral Stylophora pistillata.

    PubMed

    Karako-Lampert, Sarit; Zoccola, Didier; Salmon-Divon, Mali; Katzenellenbogen, Mark; Tambutté, Sylvie; Bertucci, Anthony; Hoegh-Guldberg, Ove; Deleury, Emeline; Allemand, Denis; Levy, Oren

    2014-01-01

    The principal architects of coral reefs are the scleractinian corals; these species are divided in two major clades referred to as "robust" and "complex" corals. Although the molecular diversity of the "complex" clade has received considerable attention, with several expressed sequence tag (EST) libraries and a complete genome sequence having been constructed, the "robust" corals have received far less attention, despite the fact that robust corals have been prominent focal points for ecological and physiological studies. Filling this gap affords important opportunities to extend these studies and to improve our understanding of the differences between the two major clades. Here, we present an EST library from Stylophora pistillata (Esper 1797) and systematically analyze the assembled transcripts compared to putative homologs from the complete proteomes of six well-characterized metazoans: Nematostella vectensis, Hydra magnipapillata, Caenorhabditis elegans, Drosophila melanogaster, Strongylocentrotus purpuratus, Ciona intestinalis and Homo sapiens. Furthermore, comparative analyses of the Stylophora pistillata ESTs were performed against several Cnidaria from the Scleractinia, Actiniaria and Hydrozoa, as well as against other stony corals separately. Functional characterization of S. pistillata transcripts into KOG/COG categories and further description of Wnt and bone morphogenetic protein (BMP) signaling pathways showed that the assembled EST library provides sufficient data and coverage. These features of this new library suggest considerable opportunities for extending our understanding of the molecular and physiological behavior of "robust" corals. PMID:24551124

  3. Habitat structure is more important than nutrient supply in modifying mussel bed assemblage in an upwelling area of the Peruvian coast

    NASA Astrophysics Data System (ADS)

    Firstater, Fausto N.; Hidalgo, Fernando J.; Lomovasky, Betina J.; Ramos, Elmer; Gamero, Patricia; Iribarne, Oscar O.

    2011-06-01

    Upwelling intensity modifies coastal primary production and influences individual traits of habitat-forming species. Along the Peruvian coast, beds of the mytilid Perumytilus purpuratus provide structurally complex habitats that harbour many organisms. We predict that in the nutrient-rich system of Central Peru, the modification of structural complexity would have stronger effects on the Perumytilus community than nutrient addition. We experimentally examined the effects of nutrient addition on the Perumytilus-dominated assemblage and we evaluated the potential effect of varying shell size on the Perumytilus-dominated assemblage. Nutrient addition to the mussel bed with slow-release fertilizers caused no changes in the total macro- and microalgal biomass and did not affect abundances or composition of the assemblage. To explore the effect of structural complexity on the Perumytilus assemblage, we manipulated mussel size with experimental bags containing small and large individuals. Predators, grazers and mobile organisms were more abundant among smaller mussels, with smaller gap volume, whereas the barnacle Jehlius cirratus was more abundant on larger mussels. In conclusion, point-source nutrient addition to the mussel bed did not enhance primary production. However, the modification of structural characteristics related to mussel size induced changes in the faunal assemblage. Thus, it seems that in this nutrient-rich system, nutrient enhancement would not significantly affect Perumytilus and its assemblage, whereas structural habitat seems to play an important role in shaping this community.

  4. Competition among Eggs Shifts to Cooperation along a Sperm Supply Gradient in an External Fertilizer.

    PubMed

    Okamoto, Daniel K

    2016-05-01

    Competition among gametes for fertilization imposes strong selection. For external fertilizers, this selective pressure extends to eggs for which spawning conditions can range from sperm limitation (competition among eggs) to sexual conflict (overabundance of competing sperm toxic to eggs). Yet existing fertilization models ignore dynamics that can alter the functional nature of gamete interactions. These factors include attraction of sperm to eggs, egg crowding effects, or other nonlinearities in per capita rates of sperm-egg interaction. Such processes potentially allow egg concentrations to drastically affect viable fertilization probabilities. I experimentally tested whether such egg effects occur, using the urchin Strongylocentrotus purpuratus, and parameterized a newly derived model of fertilization dynamics and existing models modified to include such interactions. The experiments revealed that at low sperm concentrations, eggs compete for sperm, while at high sperm concentrations, eggs cooperatively reduce abnormal fertilization (a proxy for polyspermy). I show that these observations are consistent with declines in the per capita rate at which sperm and eggs interact as eggs increase in density. The results suggest a fitness trade-off of egg release during spawning: as sperm range from scarce to superabundant, interactions among eggs transition from highly competitive to cooperative in terms of viable fertilization probabilities. PMID:27105001

  5. Anisotropic mechanical behaviors and their structural dependences of crossed-lamellar structure in a bivalve shell.

    PubMed

    Jiao, D; Liu, Z Q; Qu, R T; Zhang, Z F

    2016-02-01

    Crossed-lamellar structure is one of the most common organizations found in mollusk shells and may serve as a natural mimetic model for designing bio-inspired synthetic materials. Nonetheless, the mechanical behaviors and corresponding mechanisms have rarely been investigated for individual macro-layer of such structure. The integrated effects of orientation and hydration also remain unclear. In this study, the mechanical behaviors and their structural dependences of pure crossed-lamellar structure in Saxidomus purpuratus shell were systematically examined by three-point bending and compression tests. Mechanical properties and fracture mechanisms were revealed to depend strongly on the orientation, hydration state and loading condition. Three basic cracking modes of inter-platelet, trans-platelet, and along the interfaces between first-order lamellae were identified, and the interfacial separation was enhanced by hydration. Macroscopic compressive fracture was accomplished through axial splitting during which multiple toughening mechanisms were activated. The competition among different cracking modes was quantitatively evaluated by analyzing their driving stresses and resistances from fundamental mechanics. This study helps to clarify the mechanical behaviors of naturally occurring crossed-lamellar structure, and accordingly, aids in designing new bio-inspired synthetic materials by mimicking it. PMID:26652438

  6. P16 is an essential regulator of skeletogenesis in the sea urchin embryo.

    PubMed

    Cheers, Melani S; Ettensohn, Charles A

    2005-07-15

    The primary mesenchyme cells (PMCs) of the sea urchin embryo undergo a dramatic sequence of morphogenetic behaviors that culminates in the formation of the larval endoskeleton. Recent studies have identified components of a gene regulatory network that underlies PMC specification and differentiation. In previous work, we identified novel gene products expressed specifically by PMCs (Illies, M.R., Peeler, M.T., Dechtiaruk, A.M., Ettensohn, C.A., 2002. Identification and developmental expression of new biomineralization proteins in the sea urchin, Strongylocentrotus purpuratus. Dev. Genes Evol. 212, 419-431). Here, we show that one of these gene products, P16, plays an essential role in skeletogenesis. P16 is not required for PMC specification, ingression, migration, or fusion, but is essential for skeletal rod elongation. We have compared the predicted sequences of P16 from two species and show that this small, acidic protein is highly conserved in both structure and function. The predicted amino acid sequence of P16 and the subcellular localization of a GFP-tagged form of the protein suggest that P16 is enriched in the plasma membrane. It may function to receive signals required for skeletogenesis or may play a more direct role in the deposition of biomineral. Finally, we place P16 downstream of Alx1 in the PMC gene network, thereby linking the network to a specific "effector" protein involved in biomineralization. PMID:15935341

  7. Early development and neurogenesis of Temnopleurus reevesii.

    PubMed

    Yaguchi, Shunsuke; Yamazaki, Atsuko; Wada, Wakana; Tsuchiya, Yasutaka; Sato, Toshihiko; Shinagawa, Hideo; Yamada, Yutaro; Yaguchi, Junko

    2015-04-01

    Sea urchins are model non-chordate deuterostomes, and studying the nervous system of their embryos can aid in the understanding of the universal mechanisms of neurogenesis. However, despite the long history of sea urchin embryology research, the molecular mechanisms of their neurogenesis have not been well investigated, in part because neurons appear relatively late during embryogenesis. In this study, we used the species Temnopleurus reevesii as a new sea urchin model and investigated the detail of its development and neurogenesis during early embryogenesis. We found that the embryos of T. reevesii were tolerant of high temperatures and could be cultured successfully at 15-30°C during early embryogenesis. At 30°C, the embryos developed rapidly enough that the neurons appeared at just after 24 h. This is faster than the development of other model urchins, such as Hemicentrotus pulcherrimus or Strongylocentrotus purpuratus. In addition, the body of the embryo was highly transparent, allowing the details of the neural network to be easily captured by ordinary epifluorescent and confocal microscopy without any additional treatments. Because of its rapid development and high transparency during embryogenesis, T. reevesii may be a suitable sea urchin model for studying neurogenesis. Moreover, the males and females are easily distinguishable, and the style of early cleavages is intriguingly unusual, suggesting that this sea urchin might be a good candidate for addressing not only neurology but also cell and developmental biology. PMID:25754419

  8. A cnidarian homologue of an insect gustatory receptor functions in developmental body patterning

    PubMed Central

    Saina, Michael; Busengdal, Henriette; Sinigaglia, Chiara; Petrone, Libero; Oliveri, Paola; Rentzsch, Fabian; Benton, Richard

    2015-01-01

    Insect Gustatory and Odorant Receptors (GRs and ORs) form a superfamily of novel transmembrane proteins, which are expressed in chemosensory neurons that detect environmental stimuli. Here we identify homologues of GRs (Gustatory receptor-like (Grl) genes) in genomes across Protostomia, Deuterostomia and non-Bilateria. Surprisingly, two Grls in the cnidarian Nematostella vectensis, NvecGrl1 and NvecGrl2, are expressed early in development, in the blastula and gastrula, but not at later stages when a putative chemosensory organ forms. NvecGrl1 transcripts are detected around the arboral pole, considered the equivalent to the head-forming region of Bilateria. Morpholino-mediated knockdown of NvecGrl1 causes developmental patterning defects of this region, leading to animals lacking the apical sensory organ. A deuterostome Grl from the sea urchin Strongylocentrotus purpuratus displays similar patterns of developmental expression. These results reveal an early evolutionary origin of the insect chemosensory receptor family, and raise the possibility that their ancestral role was in embryonic development. PMID:25692633

  9. Patterns of Mass Mortality among Rocky Shore Invertebrates across 100 km of Northeastern Pacific Coastline.

    PubMed

    Jurgens, Laura J; Rogers-Bennett, Laura; Raimondi, Peter T; Schiebelhut, Lauren M; Dawson, Michael N; Grosberg, Richard K; Gaylord, Brian

    2015-01-01

    Mass mortalities in natural populations, particularly those that leave few survivors over large spatial areas, may cause long-term ecological perturbations. Yet mass mortalities may remain undocumented or poorly described due to challenges in responding rapidly to unforeseen events, scarcity of baseline data, and difficulties in quantifying rare or patchily distributed species, especially in remote or marine systems. Better chronicling the geographic pattern and intensity of mass mortalities is especially critical in the face of global changes predicted to alter regional disturbance regimes. Here, we couple replicated post-mortality surveys with preceding long-term surveys and historical data to describe a rapid and severe mass mortality of rocky shore invertebrates along the north-central California coast of the northeastern Pacific Ocean. In late August 2011, formerly abundant intertidal populations of the purple sea urchin (Strongylocentrotus purpuratus, a well-known ecosystem engineer), and the predatory six-armed sea star (Leptasterias sp.) were functionally extirpated from ~100 km of coastline. Other invertebrates, including the gumboot chiton (Cryptochiton stelleri) the ochre sea star (Pisaster ochraceus), and subtidal populations of purple sea urchins also exhibited elevated mortality. The pattern and extent of mortality suggest the potential for long-term population, community, and ecosystem consequences, recovery from which may depend on the different dispersal abilities of the affected species. PMID:26039349

  10. Validating the Assumption of Annual Shell Ring Deposition in Freshwater Mussels

    NASA Astrophysics Data System (ADS)

    Commens, A. M.; Haag, W. R.

    2005-05-01

    We evaluated the assumption of annual shell ring deposition by freshwater mussels in three rivers (Little Tallahatchie, Mississippi, 2000 and 2003; Sipsey, Alabama, 2000; St. Frances, Arkansas, 2003) using 14 species (Amblema plicata, Elliptio arca, Fusconaia cerina, F. flava, Lampsilis cardium, L. teres, Leptodea fragilis, Obliquaria reflexa, Potamilus purpuratus, Quadrula asperata, Q. pustulosa, Q. quadrula, Q, rumphiana, Tritogonia verrucosa). In 2000 we filed a notch in the shell margin, returned animals to the water, and retrieved them one year later. In 2003 we measured shell length and affixed numbered tags, returned animals to the water, and retrieved them one year later. After retrieval, we examined external and internal shell rings using thin-sections. For both methods, all species at all sites grew and deposited a single winter rest line between times of initial and final collection. Handling produced a conspicuous disturbance ring in all species. Characteristics of annual rings, disturbance rings, and false annuli differed qualitatively. Growth of some individuals was reduced after handling but negative growth occurred only in a few large individuals. Annual shell ring formation occurs consistently across species, space, and time. Formation of disturbance rings associated with collection indicates that mussels are extremely sensitive to handling.

  11. Selective accumulation of germ-line associated gene products in early development of the sea star and distinct differences from germ-line development in the sea urchin

    PubMed Central

    Fresques, Tara; Zazueta-Novoa, Vanesa; Reich, Adrian; Wessel, Gary M.

    2014-01-01

    Background Echinodermata is a diverse Phylum, a sister group to chordates, and contains diverse organisms that may be useful to understand varied mechanisms of germ-line specification. Results We tested 23 genes in development of the sea star Patiria miniata that fall into five categories: 1) Conserved germ-line factors; 2) Genes involved in the inductive mechanism of germ-line specification; 3) Germ-line associated genes; 4) Molecules involved in left-right asymmetry; and 5) Genes involved in regulation and maintenance of the genome during early embryogenesis. Overall, our results support the contention that the posterior enterocoel is a source of the germ line in the sea star P. miniata. Conclusion The germ line in this organism appears to be specified late in embryogenesis, and in a pattern more consistent with inductive interactions amongst cells. This is distinct from the mechanism seen in sea urchins, a close relative of the sea star clad. We propose that P. miniata may serve as a valuable model to study inductive mechanisms of germ-cell specification and when compared to germ-line formation in the sea urchin S. purpuratus may reveal developmental transitions that occur in the evolution of inherited and inductive mechanisms of germ-line specification. PMID:24038550

  12. The echinoderm adhesome

    PubMed Central

    Whittaker, Charles A.; Bergeron, Karl-Frederik; Whittle, James; Brandhorst, Bruce P.; Burke, Robert D.; Hynes, Richard O.

    2013-01-01

    Although the development of sea urchin embryos has been studied extensively and clearly involves both cell adhesion and cell migration, rather little is known about the adhesion receptors and extracellular matrix molecules involved. The completion of the genome of Strongylocentrotus purpuratus allows a comprehensive survey of the complement of cell-cell and cell-matrix adhesion molecules in this organism. Furthermore, the phylogenetic position of echinoderms offers the opportunity to compare the complement of adhesion proteins between protostome and deuterostome invertebrates and between invertebrate and vertebrate deuterostomes. Many aspects of development and cell interactions differ among these different taxa and it is likely that analysis of the spectrum of adhesion receptors and extracellular matrix proteins can open up new insights into which molecules have evolved to suit particular developmental processes. In this paper, we report the results of an initial analysis along these lines. The echinoderm adhesome (complement of adhesion-related genes/proteins) is similar overall to that of other invertebrates although there are significant deuterostome-specific innovations and some interesting features previously thought to be chordate- or vertebrate-specific. PMID:16950242

  13. Seawi—a sea urchin piwi/argonaute family member is a component of MT-RNP complexes

    PubMed Central

    RODRIGUEZ, ALEXIS J.; SEIPEL, SUSAN A.; HAMILL, DANIELLE R.; ROMANCINO, DANIELE P.; DI CARLO, MARTA; SUPRENANT, KATHY A.; BONDER, EDWARD M.

    2005-01-01

    The piwi/argonaute family of proteins is involved in key developmental processes such as stem cell maintenance and axis specification through molecular mechanisms that may involve RNA silencing. Here we report on the cloning and characterization of the sea urchin piwi/argonaute family member seawi. Seawi is a major component of microtubule-ribonucleoprotein (MT-RNP) complexes isolated from two different species of sea urchin, Strongylocentrotus purpuratus and Paracentrotus lividus. Seawi co-isolates with purified ribosomes, cosediments with 80S ribosomes in sucrose density gradients, and binds microtubules. Seawi possesses the RNA binding motif common to piwi family members and binds P. lividus bep4 mRNA, a transcript that co-isolates with MT-RNP complexes and whose translation product has been shown to play a role in patterning the animal–vegetal axis. Indirect immunofluorescence studies localized seawi to the cortex of unfertilized eggs within granule-like particles, the mitotic spindle during cell division, and the small micromeres where its levels were enriched during the early cleavage stage. Lastly, we discuss how seawi, as a piwi/argonaute family member, may play a fundamentally important role in sea urchin animal–vegetal axis formation and stem cell maintenance. PMID:15840816

  14. The cis-regulatory system of the tbrain gene: alternative use of multiple modules to promote skeletogenic expression in the sea urchin embryo

    PubMed Central

    Wahl, Mary E.; Hahn, Julie; Gora, Kasia; Davidson, Eric H.; Oliveri, Paola

    2009-01-01

    The genomic cis-regulatory systems controlling regulatory gene expression usually include multiple modules. The regulatory output of such systems at any given time depends on which module is directing the function of the basal transcription apparatus, and ultimately on the transcription factor inputs into that module. Here we examine regulation of the S. purpuratus tbrain gene, a required activator of the skeletogenic specification state in the lineage descendant from the embryo micromeres. Alternate cis-regulatory modules were found to convey skeletogenic expression in reporter constructs. To determine their relative developmental functions in context, we made use of recombineered BAC constructs containing a GFP reporter, and of derivatives from which specific modules had been deleted. The outputs of the various constructs were observed spatially by GFP fluorescence and quantitatively over time by QPCR. In the context of the complete genomic locus, early skeletogenic expression is controlled by an intron enhancer plus a proximal region containing a HesC site as predicted from network analysis. From ingression onward, however, a dedicated distal module utilizing positive Ets1/2 inputs contributes to definitive expression in the skeletogenic mesenchyme. This module also mediates a newly-discovered negative Erg input which excludes non-skeletogenic mesodermal expression. PMID:19679118

  15. Phylogenomic resolution of the hemichordate and echinoderm clade.

    PubMed

    Cannon, Johanna T; Kocot, Kevin M; Waits, Damien S; Weese, David A; Swalla, Billie J; Santos, Scott R; Halanych, Kenneth M

    2014-12-01

    Ambulacraria, comprising Hemichordata and Echinodermata, is closely related to Chordata, making it integral to understanding chordate origins and polarizing chordate molecular and morphological characters. Unfortunately, relationships within Hemichordata and Echinodermata have remained unresolved, compromising our ability to extrapolate findings from the most closely related molecular and developmental models outside of Chordata (e.g., the acorn worms Saccoglossus kowalevskii and Ptychodera flava and the sea urchin Strongylocentrotus purpuratus). To resolve long-standing phylogenetic issues within Ambulacraria, we sequenced transcriptomes for 14 hemichordates as well as 8 echinoderms and complemented these with existing data for a total of 33 ambulacrarian operational taxonomic units (OTUs). Examination of leaf stability values revealed rhabdopleurid pterobranchs and the enteropneust Stereobalanus canadensis were unstable in placement; therefore, analyses were also run without these taxa. Analyses of 185 genes resulted in reciprocal monophyly of Enteropneusta and Pterobranchia, placed the deep-sea family Torquaratoridae within Ptychoderidae, and confirmed the position of ophiuroid brittle stars as sister to asteroid sea stars (the Asterozoa hypothesis). These results are consistent with earlier perspectives concerning plesiomorphies of Ambulacraria, including pharyngeal gill slits, a single axocoel, and paired hydrocoels and somatocoels. The resolved ambulacrarian phylogeny will help clarify the early evolution of chordate characteristics and has implications for our understanding of major fossil groups, including graptolites and somasteroideans. PMID:25454590

  16. Intrinsic hierarchical structural imperfections in a natural ceramic of bivalve shell with distinctly graded properties

    PubMed Central

    Jiao, Da; Liu, Zengqian; Zhang, Zhenjun; Zhang, Zhefeng

    2015-01-01

    Despite the extensive investigation on the structure of natural biological materials, insufficient attention has been paid to the structural imperfections by which the mechanical properties of synthetic materials are dominated. In this study, the structure of bivalve Saxidomus purpuratus shell has been systematically characterized quantitatively on multiple length scales from millimeter to sub-nanometer. It is revealed that hierarchical imperfections are intrinsically involved in the crossed-lamellar structure of the shell despite its periodically packed platelets. In particular, various favorable characters which are always pursued in synthetic materials, e.g. nanotwins and low-angle misorientations, have been incorporated herein. The possible contributions of these imperfections to mechanical properties are further discussed. It is suggested that the imperfections may serve as structural adaptations, rather than detrimental defects in the real sense, to help improve the mechanical properties of natural biological materials. This study may aid in understanding the optimizing strategies of structure and properties designed by nature, and accordingly, provide inspiration for the design of synthetic materials. PMID:26198844

  17. Marine organism cell biology and regulatory sequence discoveryin comparative functional genomics.

    PubMed

    Barnes, David W; Mattingly, Carolyn J; Parton, Angela; Dowell, Lori M; Bayne, Christopher J; Forrest, John N

    2004-10-01

    The use of bioinformatics to integrate phenotypic and genomic data from mammalian models is well established as a means of understanding human biology and disease. Beyond direct biomedical applications of these approaches in predicting structure-function relationships between coding sequences and protein activities, comparative studies also promote understanding of molecular evolution and the relationship between genomic sequence and morphological and physiological specialization. Recently recognized is the potential of comparative studies to identify functionally significant regulatory regions and to generate experimentally testable hypotheses that contribute to understanding mechanisms that regulate gene expression, including transcriptional activity, alternative splicing and transcript stability. Functional tests of hypotheses generated by computational approaches require experimentally tractable in vitro systems, including cell cultures. Comparative sequence analysis strategies that use genomic sequences from a variety of evolutionarily diverse organisms are critical for identifying conserved regulatory motifs in the 5'-upstream, 3'-downstream and introns of genes. Genomic sequences and gene orthologues in the first aquatic vertebrate and protovertebrate organisms to be fully sequenced (Fugu rubripes, Ciona intestinalis, Tetraodon nigroviridis, Danio rerio) as well as in the elasmobranchs, spiny dogfish shark (Squalus acanthias) and little skate (Raja erinacea), and marine invertebrate models such as the sea urchin (Strongylocentrotus purpuratus) are valuable in the prediction of putative genomic regulatory regions. Cell cultures have been derived for these and other model species. Data and tools resulting from these kinds of studies will contribute to understanding transcriptional regulation of biomedically important genes and provide new avenues for medical therapeutics and disease prevention. PMID:19003267

  18. Localization and Substrate Selectivity of Sea Urchin Multidrug (MDR) Efflux Transporters*

    PubMed Central

    Gökirmak, Tufan; Campanale, Joseph P.; Shipp, Lauren E.; Moy, Gary W.; Tao, Houchao; Hamdoun, Amro

    2012-01-01

    In this study, we cloned, expressed and functionally characterized Stronglycentrotus purpuratus (Sp) ATP-binding cassette (ABC) transporters. This screen identified three multidrug resistance (MDR) transporters with functional homology to the major types of MDR transporters found in humans. When overexpressed in embryos, the apical transporters Sp-ABCB1a, ABCB4a, and ABCG2a can account for as much as 87% of the observed efflux activity, providing a robust assay for their substrate selectivity. Using this assay, we found that sea urchin MDR transporters export canonical MDR susbtrates such as calcein-AM, bodipy-verapamil, bodipy-vinblastine, and mitoxantrone. In addition, we characterized the impact of nonconservative substitutions in the primary sequences of drug binding domains of sea urchin versus murine ABCB1 by mutation of Sp-ABCB1a and treatment of embryos with stereoisomeric cyclic peptide inhibitors (QZ59 compounds). The results indicated that two substitutions in transmembrane helix 6 reverse stereoselectivity of Sp-ABCB1a for QZ59 enantiomers compared with mouse ABCB1a. This suggests that subtle changes in the primary sequence of transporter drug binding domains could fine-tune substrate specificity through evolution. PMID:23124201

  19. ATP-binding cassette (ABC) transporter expression and localization in sea urchin development

    PubMed Central

    Shipp, Lauren E.; Hamdoun, Amro

    2012-01-01

    Background ATP-binding cassette (ABC) transporters are membrane proteins that regulate intracellular concentrations of myriad compounds and ions. There are >100 ABC transporter predictions in the Strongylocentrotus purpuratus genome, including 40 annotated ABCB, ABCC, and ABCG “multidrug efflux” transporters. Despite the importance of multidrug transporters for protection and signaling, their expression patterns have not been characterized in deuterostome embryos. Results Sea urchin embryos expressed 20 ABCB, ABCC, and ABCG transporter genes in the first 58 hours of development, from unfertilized egg to early prism. We quantified transcripts of ABCB1a, ABCB4a, ABCC1, ABCC5a, ABCC9a, and ABCG2b, and found that ABCB1a mRNA was 10–100 times more abundant than other transporter mRNAs. In situ hybridization showed ABCB1a was expressed ubiquitously in embryos, while ABCC5a was restricted to secondary mesenchyme cells and their precursors. Fluorescent protein fusions showed localization of ABCB1a on apical cell surfaces, and ABCC5a on basolateral surfaces. Conclusions Embryos utilize many ABC transporters with predicted functions in cell signaling, lysosomal and mitochondrial homeostasis, potassium channel regulation, pigmentation, and xenobiotic efflux. Detailed characterization of ABCB1a and ABCC5a revealed that they have different temporal and spatial gene expression profiles and protein localization patterns that correlate to their predicted functions in protection and development, respectively. PMID:22473856

  20. Evolution of thrombin and other hemostatic proteases by survey of protochordate, hemichordate, and echinoderm genomes.

    PubMed

    Ponczek, Michal B; Bijak, Michal Z; Nowak, Pawel Z

    2012-06-01

    Protochordate genomes enable a prevalence of hemostasis evolution. Broad searches were performed for homologs of human serine proteases of hemostasis on the genomes of Branchiostoma floridae, Saccoglossus kowalevskii, and Strongylocentrotus purpuratus. Sequences were analyzed by multiple bioinformatic tools. The survey revealed numerous homologous components. Amphioxus was rich in some serine proteases not accompanied by gamma-carboxyglutamic or kringle domains similar more to thrombin than to other coagulation factors. The serine proteases found in amphioxus exhibited the attributes similar to those of thrombin by phylogeny relationships, sequence conservation, gene synteny, spatial structure, and ligand docking. A few plasminogen- and plasminogen activators-like proteases with kringles were also present. Those serine proteases demonstrated the greatest proximity rather to plasminogen or plasminogen activators than to thrombin. Searching for homologs of serine protease hemostatic factors in acorn worm and sea urchin revealed several components similar to those found in amphioxus. Hypothetically, the common ancestor of chordates had three separate serine proteases that evolved independently into immunoglobulin-like and kringle proteases in lancelets, and prothrombin, plasminogen activators, and plasminogen in vertebrates. Ancestral proteases evolved in vertebrates into hemostasis factors after merging the proper N-terminal domains and duplications. PMID:22752046

  1. Identification and comparative analysis of complement C3-associated microRNAs in immune response of Apostichopus japonicus by high-throughput sequencing.

    PubMed

    Zhong, Lei; Zhang, Feng; Zhai, Yu; Cao, Yanhui; Zhang, Si; Chang, Yaqing

    2015-01-01

    MicroRNAs (miRNAs) are important effectors in mediating host-pathogen interaction. In this report, coelomocytes miRNA libraries of three Japanese sea cucumbers Apostichopus japonicus were built by Illumina(®) Hiseq2000 from different time points after lipopolysaccharide challenge (at time 0 h, 6 h and 12 h). The clean data received from high throughput sequencing were used to sequences analysis. Referenced to the Strongylocentrotus purpuratus genome, 38 conserved miRNAs were found, and three miRNA candidates were predicted by software. According to the evidence resulting from the expression of AjC3, expressing levels of spu-miR-133, spu-miR-137 and spu-miR-2004 altered along with the expression of AjC3 changing at different time points after LPS injection. Thus, we speculated that the three miRNAs may have influence on A. japonicus complement C3. The spu-miR-137 and miR-137 gene family in miRBase were analyzed by bioinformatics. There is an obvious discrepancy between invertebrates and vertebrates. The first and ninth nucleotides in invertebrate miR-137 are offset compared vertebrate miR-137. Importantly, this is the first attempt to map the stage of immune response regulome in echinoderms, which might be considered as information for elucidating the intrinsic mechanism underlying the immune system in this species. PMID:26634300

  2. Identification of Cytochrome P450 ( CYP) genes in Zhikong scallop ( Chlamys farreri)

    NASA Astrophysics Data System (ADS)

    Guo, Huihui; Bao, Zhenmin; Du, Huixia; Zhang, Lingling; Wang, Shi; Sun, Luyang; Mou, Xiaoyu; Hu, Xiaoli

    2013-03-01

    Cytochrome P450 ( CYP) superfamily is one of the membership largest and function most diverse protein superfamily recogniozed among living beings. Members of this superfamily were further assigned to different families and subfamilies based on their amino acid similarities. According to their phylogenetic relationships, the CYP genes which likely diverged from common ancestor gene and may share common functions were grouped into one clan. Widely distributing scallops are a group of the most conspicuous bivalve; however the studies on their CYP is acarce. In this study, we searched the genome and expressed sequence tags of Zhikong scallop ( Chlamys farreri) for CYP genes. In total, 88 non-redundant CYP were identified, which were homed in 13 CYPs gene families. Phylogenetic analysis divided these genes into 4 CYP clans. As in deuterostomes, Clan 2 was the largest, which contained 33 genes belonging to CYP1, CYP2, CYP17 and CYP356 families. Clan 3 contgained 19 genes belonging to CYP3, CYP5 and CYP30 families. Clan 4 contained 23 genes, all belonging to CYP4 family. The mitochondrial CYP clan contained 9 genes belonging to CYP10 and CYP24 families. In comparison, protostomes ( C. farreri, D. pluex, D. melanogaster) contained more CYP genes than deuterostomes ( S. purpuratus and vertebrates) in Clan 2 but less genes in Clan 3 and Clan 4. Our findings will aid to deciphering CYP function and evolution in scallops and bivalves.

  3. Cloning of two sea urchin DNA-binding proteins involved in mitochondrial DNA replication and transcription.

    PubMed

    Loguercio Polosa, Paola; Megli, Fiammetta; Di Ponzio, Barbara; Gadaleta, Maria Nicola; Cantatore, Palmiro; Roberti, Marina

    2002-03-01

    The cloning of the cDNA for two mitochondrial proteins involved in sea urchin mtDNA replication and transcription is reported here. The cDNA for the mitochondrial D-loop binding protein (mtDBP) from the sea urchin Strongylocentrotus purpuratus has been cloned by a polymerase chain reaction-based approach. The protein displays a very high similarity with the Paracentrotus lividus homologue as it contains also the two leucine zipper-like domains which are thought to be involved in intramolecular interactions needed to expose the two DNA binding domains in the correct position for contacting DNA. The cDNA for the mitochondrial single-stranded DNA-binding protein (mtSSB) from P. lividus has been also cloned by a similar approach. The precursor protein is 146 amino acids long with a presequence of 16 residues. The deduced amino acid sequence shows the highest homology with the Xenopus laevis protein and the lowest with the Drosophila mtSSB. The computer modeling of the tertiary structure of P. lividus mtSSB shows a structure very similar to that experimentally determined for human mtSSB, with the conservation of the main residues involved in protein tetramerization and in DNA binding. PMID:11943466

  4. cis-Regulatory control of the initial neurogenic pattern of onecut gene expression in the sea urchin embryo.

    PubMed

    Barsi, Julius C; Davidson, Eric H

    2016-01-01

    Specification of the ciliated band (CB) of echinoid embryos executes three spatial functions essential for postgastrular organization. These are establishment of a band about 5 cells wide which delimits and bounds other embryonic territories; definition of a neurogenic domain within this band; and generation within it of arrays of ciliary cells that bear the special long cilia from which the structure derives its name. In Strongylocentrotus purpuratus the spatial coordinates of the future ciliated band are initially and exactly determined by the disposition of a ring of cells that transcriptionally activate the onecut homeodomain regulatory gene, beginning in blastula stage, long before the appearance of the CB per se. Thus the cis-regulatory apparatus that governs onecut expression in the blastula directly reveals the genomic sequence code by which these aspects of the spatial organization of the embryo are initially determined. We screened the entire onecut locus and its flanking region for transcriptionally active cis-regulatory elements, and by means of BAC recombineered deletions identified three separated and required cis-regulatory modules that execute different functions. The operating logic of the crucial spatial control module accounting for the spectacularly precise and beautiful early onecut expression domain depends on spatial repression. Previously predicted oral ectoderm and aboral ectoderm repressors were identified by cis-regulatory mutation as the products of goosecoid and irxa genes respectively, while the pan-ectodermal activator SoxB1 supplies a transcriptional driver function. PMID:26522848

  5. Toxicity of sediments and interstitial waters form the Southern California Bight

    SciTech Connect

    Bay, S.; Greenstein, D.; Brown, J.; Jirik, A.

    1995-12-31

    The toxicity of 72 sediment samples collected during the EMAP Southern California Bight Pilot Project (SCBPP) was measured. Sediments from the mainland shelf between Point Conception and the Mexican border were collected from various depths and tested for toxicity using two methods. The toxicity of bulk sediment was measured using a 10-day amphipod (Ampelisca abdita) survival test. Interstitial water was also extracted from the samples and tested for toxicity using a 72-hour sea urchin (Strongylocentrotus purpuratus) embryo development test. Amphipod survival was high (> 80%) at all stations tested, although several sites near large sewage outfalls had statistically significant reductions in survival. No interference related to grain size variation was observed with the amphipod test. Most of the interstitial water samples produced abnormal sea urchin embryo development. Effects were not related to the presumed level of sediment contamination, but rather to ammonia concentration in virtually all cases. The impacts of sample handling procedures and ammonia on sediment toxicity data interpretation will be discussed.

  6. Biological testing of sediment for the Olympia Harbor Navigation Improvement Project, 1988: Geoduck, amphipod, and echinoderm bioassays

    SciTech Connect

    Ward, J.A.; Word, J.Q.; Antrim, L.D.

    1989-05-01

    The Olympia Harbor Navigation Improvement Project requires the dredging of approximately 330,000 cubic yards (cy) of sediment from the harbor entrance channel and 205,185 cy from the turning basin. Puget Sound Dredged Disposal Analysis (PSDDA) partial characterization studies were used to plan a full sediment characterization in which chemical analyses and biological testing of sediments evaluated the suitability of the dredged material for unconfined, open-water disposal. The US Army Corps of Engineers (COE), Seattle District, contracted with NOAA/NMFS, Environmental Conservation Division, to perform the chemical analysis and Microtox bioassay tests, and with the Battelle/Marine Sciences Laboratory (MSL) in Sequim to perform flow-through solid-phase bioassays utilizing juvenile (8 to 10 mm) geoduck clams, Panopea generosa, and static solid phase bioassays using the phoxocephalid amphipod, Rhepoxynius abronius, developing embryos and gametes of the purple sea urchin, Strongylocentrotus purpuratus, and the larvae of the Pacific oyster Crassostrea gigas. When the results of the biological tests were evaluated under PSDDA guidelines, it was found that all the tested sediment treatments from Olympia Harbor are suitable for unconfined open-water disposal. 14 refs., 12 figs., 3 tabs.

  7. Control of protein synthesis in cell-free extracts of sea urchin embryos

    SciTech Connect

    Hansen, L.J.; Huang, W.I.; Jagus, R.

    1986-05-01

    Although the increase in protein synthesis that occurs after fertilization of sea urchin eggs results from increased utilization of stored maternal mRNA, the underlying mechanism is unknown. The authors have prepared cell-free extracts from S.purpuratus and A.puctulata unfertilized eggs and 2-cell embryos that retain the protein synthetic differences observed in vivo. The method is based on that of Dr. Alina Lopo. /sup 35/S methionine incorporation is linear during a 30 min incubation and is 10-20 fold higher in extracts from 2-cell embryos than unfertilized eggs. Addition of purified mRNA does not stimulate these systems, suggesting a regulatory mechanism other than mRNA masking. Addition of rabbit reticulocyte ribosomal salt wash stimulated protein synthesis in extracts from eggs but not embryos, suggesting deficiencies in translational components in unfertilized eggs. Mixing of egg and embryo lysates indicated the presence of a weak protein synthesis inhibitor in eggs. Translational control in developing sea urchin embryos thus appears to be complex, involving both stimulatory and inhibitory factors.

  8. A recombinant Sp185/333 protein from the purple sea urchin has multitasking binding activities towards certain microbes and PAMPs.

    PubMed

    Lun, Cheng Man; Schrankel, Catherine S; Chou, Hung-Yen; Sacchi, Sandro; Smith, L Courtney

    2016-08-01

    The purple sea urchin, Strongylocentrotus purpuratus, possesses a sophisticated innate immune system that responds to microbes effectively by swift expression of the highly diverse Sp185/333 gene family. The Sp185/333 proteins are predicted to have anti-pathogen functions based on inducible gene expression and their significant sequence diversity. Sp185/333 proteins are all predicted to be intrinsically disordered and do not exhibit sequence similarities to other known proteins. To test the anti-pathogen hypothesis, a recombinant Sp185/333 protein, rSp0032, was evaluated and found to exhibit specific binding to marine Vibrio diazotrophicus and to Saccharomyces cerevisiae, but not to two Bacillus species. rSp0032 also binds to LPS, β-1,3-glucan and flagellin but not to peptidoglycan. rSp0032 binding to LPS can be competed by LPS, β-1,3-glucan and flagellin but not by peptidoglycan. We speculate that the predicted intrinsically disordered structure of rSp0032 may adapt to different conformations in binding to a limited number of PAMPs and pathogens. Given that rSp0032 binds to a range of targets, and that up to 260 different Sp185/333 proteins can be expressed per individual sea urchin, this family of immune response proteins may facilitate effective host protection against a broad array of potential pathogens encountered in the marine environment. PMID:27020848

  9. Maintenance of somatic tissue regeneration with age in short- and long-lived species of sea urchins.

    PubMed

    Bodnar, Andrea G; Coffman, James A

    2016-08-01

    Aging in many animals is characterized by a failure to maintain tissue homeostasis and the loss of regenerative capacity. In this study, the ability to maintain tissue homeostasis and regenerative potential was investigated in sea urchins, a novel model to study longevity and negligible senescence. Sea urchins grow indeterminately, regenerate damaged appendages and reproduce throughout their lifespan and yet different species are reported to have very different life expectancies (ranging from 4 to more than 100 years). Quantitative analyses of cell proliferation and apoptosis indicated a low level of cell turnover in tissues of young and old sea urchins of species with different lifespans (Lytechinus variegatus, Strongylocentrotus purpuratus and Mesocentrotus franciscanus). The ability to regenerate damaged tissue was maintained with age as assessed by the regrowth of amputated spines and tube feet (motor and sensory appendages). Expression of genes involved in cell proliferation (pcna), telomere maintenance (tert) and multipotency (seawi and vasa) was maintained with age in somatic tissues. Immunolocalization of the Vasa protein to areas of the tube feet, spines, radial nerve, esophagus and a sub-population of circulating coelomocytes suggests the presence of multipotent cells that may play a role in normal tissue homeostasis and the regenerative potential of external appendages. The results indicate that regenerative potential was maintained with age regardless of lifespan, contrary to the expectation that shorter lived species would invest less in maintenance and repair. PMID:27095483

  10. Spatial expression of Hox cluster genes in the ontogeny of a sea urchin

    NASA Technical Reports Server (NTRS)

    Arenas-Mena, C.; Cameron, A. R.; Davidson, E. H.

    2000-01-01

    The Hox cluster of the sea urchin Strongylocentrous purpuratus contains ten genes in a 500 kb span of the genome. Only two of these genes are expressed during embryogenesis, while all of eight genes tested are expressed during development of the adult body plan in the larval stage. We report the spatial expression during larval development of the five 'posterior' genes of the cluster: SpHox7, SpHox8, SpHox9/10, SpHox11/13a and SpHox11/13b. The five genes exhibit a dynamic, largely mesodermal program of expression. Only SpHox7 displays extensive expression within the pentameral rudiment itself. A spatially sequential and colinear arrangement of expression domains is found in the somatocoels, the paired posterior mesodermal structures that will become the adult perivisceral coeloms. No such sequential expression pattern is observed in endodermal, epidermal or neural tissues of either the larva or the presumptive juvenile sea urchin. The spatial expression patterns of the Hox genes illuminate the evolutionary process by which the pentameral echinoderm body plan emerged from a bilateral ancestor.

  11. Cell mediated immune response of the Mediterranean sea urchin Paracentrotus lividus after PAMPs stimulation.

    PubMed

    Romero, A; Novoa, B; Figueras, A

    2016-09-01

    The Mediterranean sea urchin (Paracentrotus lividus) is of great ecological and economic importance for the European aquaculture. Yet, most of the studies regarding echinoderm's immunological defense mechanisms reported so far have used the sea urchin Strongylocentrotus purpuratus as a model, and information on the immunological defense mechanisms of Paracentrotus lividus and other sea urchins, is scarce. To remedy this gap in information, in this study, flow cytometry was used to evaluate several cellular immune mechanisms, such as phagocytosis, cell cooperation, and ROS production in P. lividus coelomocytes after PAMP stimulation. Two cell populations were described. Of the two, the amoeboid-phagocytes were responsible for the phagocytosis and ROS production. Cooperation between amoeboid-phagocytes and non-adherent cells resulted in an increased phagocytic response. Stimulation with several PAMPs modified the phagocytic activity and the production of ROS. The premise that the coelomocytes were activated by the bacterial components was confirmed by the expression levels of two cell mediated immune genes: LPS-Induced TNF-alpha Factor (LITAF) and macrophage migration inhibitory factor (MIF). These results have helped us understand the cellular immune mechanisms in P. lividus and their modulation after PAMP stimulation. PMID:27113124

  12. Transformation and Crystallization Energetics of Synthetic and Biogenic Amorphous Calcium Carbonate

    SciTech Connect

    Radha, A. V.; Forbes, Tori Z.; Killian, Christopher E.; Gilbert, P.U.P.A; Navrotsky, Alexandra

    2010-01-01

    Amorphous calcium carbonate (ACC) is a metastable phase often observed during low temperature inorganic synthesis and biomineralization. ACC transforms with aging or heating into a less hydrated form, and with time crystallizes to calcite or aragonite. The energetics of transformation and crystallization of synthetic and biogenic (extracted from California purple sea urchin larval spicules, Strongylocentrotus purpuratus) ACC were studied using isothermal acid solution calorimetry and differential scanning calorimetry. Transformation and crystallization of ACC can follow an energetically downhill sequence: more metastable hydrated ACC → less metastable hydrated ACC→anhydrous ACC ~ biogenic anhydrous ACC→vaterite → aragonite → calcite. In a given reaction sequence, not all these phases need to occur. The transformations involve a series of ordering, dehydration, and crystallization processes, each lowering the enthalpy (and free energy) of the system, with crystallization of the dehydrated amorphous material lowering the enthalpy the most. ACC is much more metastable with respect to calcite than the crystalline polymorphs vaterite or aragonite. The anhydrous ACC is less metastable than the hydrated, implying that the structural reorganization during dehydration is exothermic and irreversible. Dehydrated synthetic and anhydrous biogenic ACC are similar in enthalpy. The transformation sequence observed in biomineralization could be mainly energetically driven; the first phase deposited is hydrated ACC, which then converts to anhydrous ACC, and finally crystallizes to calcite. The initial formation of ACC may be a first step in the precipitation of calcite under a wide variety of conditions, including geological CO₂ sequestration.

  13. Genome-wide identification of enhancer elements.

    PubMed

    Tulin, Sarah; Barsi, Julius C; Bocconcelli, Carlo; Smith, Joel

    2016-01-01

    We present a prospective genome-wide regulatory element database for the sea urchin embryo and the modified chromosome capture-related methodology used to create it. The method we developed is termed GRIP-seq for genome-wide regulatory element immunoprecipitation and combines features of chromosome conformation capture, chromatin immunoprecipitation, and paired-end next-generation sequencing with molecular steps that enrich for active cis-regulatory elements associated with basal transcriptional machinery. The first GRIP-seq database, available to the community, comes from S. purpuratus 24 hpf embryos and takes advantage of the extremely well-characterized cis-regulatory elements in this system for validation. In addition, using the GRIP-seq database, we identify and experimentally validate a novel, intronic cis-regulatory element at the onecut locus. We find GRIP-seq signal sensitively identifies active cis-regulatory elements with a high signal-to-noise ratio for both distal and intronic elements. This promising GRIP-seq protocol has the potential to address a rate-limiting step in resolving comprehensive, predictive network models in all systems. PMID:27389984

  14. Genotoxic and developmental effects in sea urchins are sensitive indicators of effects of genotoxic chemicals

    SciTech Connect

    Anderson, S.L. . Energy and Environment Division); Hose, J.E. . Dept. of Biology); Knezovich, J.P. . Health and Ecological Assessment Division)

    1994-07-01

    Purple sea urchin (Strongylocentrotus purpuratus) gametes and embryos were exposed to three known mutagenic chemicals (phenol, benzidine,and pentachlorophenol) over concentration ranges bracketing the effect levels for fertilization success. Normal development and cytogenetic effects (anaphase aberrations) were assessed after the cultures were allowed to develop for 48 h. Using radiolabeled chemicals, the authors also characterized concentrations in the test water as well as doses in the embryos following 2- and 48-h exposures. The authors observed dose responses for all chemicals and all responses, except for phenol, which showed no significant effect on development. Fertilization success was never the most sensitive end point. anaphase aberrations were the most sensitive response for phenol, with an LOEC of 2.5 mg/L exposure concentration. Anaphase aberrations and development were equivalent in sensitivity for benzidine within the tested dose range, and an LOEC of <0.1 mg/L was observed. Development was the most sensitive reasons for pentachlorophenol (LOEC 1 mg/L). the LOEC values for this study were generally lower than comparable data for aquatic life or human health protection. The authors conclude that genotoxicity and development evaluations should be included in environmental management applications and that tests developed primarily for human health protection do not reliably predict the effects of toxic substances on aquatic life.

  15. [Expression of SM30 (A-F) Genes Encoding Spicule Matrix Proteins in Intact and Damaged Sea Urchin Strongylocentrotus intermedius (A. Agassiz, 1863) at the Six-Arm Pluteus].

    PubMed

    Sharmankina, V V; Kiselev, K V

    2016-03-01

    In this study we investigated expression of the SM30(A-F) gene family encoding Strongylocentrotus intermedius spicule matrix proteins during the normal and regenerative pluteus II stage (three pairs of arms). We found that SiSM30A and SiSM30B genes are expressed at high levels in the normal pluteus II sea urchin. SiSM30A is expression was also significantly upregulated in the reparative pluteus II stage 3 hours after damage. Conversely, SiSM30B was downregulated during the reparative pluteus II stage. Our findings reveal a substantial similarity between the activity of SiSM30A and SiSM30B activity in the processes of regenerative growth during the pluteus II stage and during normal development at the prism stage in Strongylocentrotus purpuratus. On the basis of our findings, we propose that normal developmental mechanisms corresponding to the preceding developmental stage are reactivated during pluteus regeneration. PMID:27281854

  16. Patterns of Mass Mortality among Rocky Shore Invertebrates across 100 km of Northeastern Pacific Coastline

    PubMed Central

    Jurgens, Laura J.; Rogers-Bennett, Laura; Raimondi, Peter T.; Schiebelhut, Lauren M.; Dawson, Michael N.; Grosberg, Richard K.; Gaylord, Brian

    2015-01-01

    Mass mortalities in natural populations, particularly those that leave few survivors over large spatial areas, may cause long-term ecological perturbations. Yet mass mortalities may remain undocumented or poorly described due to challenges in responding rapidly to unforeseen events, scarcity of baseline data, and difficulties in quantifying rare or patchily distributed species, especially in remote or marine systems. Better chronicling the geographic pattern and intensity of mass mortalities is especially critical in the face of global changes predicted to alter regional disturbance regimes. Here, we couple replicated post-mortality surveys with preceding long-term surveys and historical data to describe a rapid and severe mass mortality of rocky shore invertebrates along the north-central California coast of the northeastern Pacific Ocean. In late August 2011, formerly abundant intertidal populations of the purple sea urchin (Strongylocentrotus purpuratus, a well-known ecosystem engineer), and the predatory six-armed sea star (Leptasterias sp.) were functionally extirpated from ~100 km of coastline. Other invertebrates, including the gumboot chiton (Cryptochiton stelleri) the ochre sea star (Pisaster ochraceus), and subtidal populations of purple sea urchins also exhibited elevated mortality. The pattern and extent of mortality suggest the potential for long-term population, community, and ecosystem consequences, recovery from which may depend on the different dispersal abilities of the affected species. PMID:26039349

  17. Coastal Upwelling Drives Intertidal Assemblage Structure and Trophic Ecology.

    PubMed

    Reddin, Carl J; Docmac, Felipe; O'Connor, Nessa E; Bothwell, John H; Harrod, Chris

    2015-01-01

    Similar environmental driving forces can produce similarity among geographically distant ecosystems. Coastal oceanic upwelling, for example, has been associated with elevated biomass and abundance patterns of certain functional groups, e.g., corticated macroalgae. In the upwelling system of Northern Chile, we examined measures of intertidal macrobenthic composition, structure and trophic ecology across eighteen shores varying in their proximity to two coastal upwelling centres, in a hierarchical sampling design (spatial scales of >1 and >10 km). The influence of coastal upwelling on intertidal communities was confirmed by the stable isotope values (δ13C and δ15N) of consumers, including a dominant suspension feeder, grazers, and their putative resources of POM, epilithic biofilm, and macroalgae. We highlight the utility of muscle δ15N from the suspension feeding mussel, Perumytilus purpuratus, as a proxy for upwelling, supported by satellite data and previous studies. Where possible, we used corrections for broader-scale trends, spatial autocorrelation, ontogenetic dietary shifts and spatial baseline isotopic variation prior to analysis. Our results showed macroalgal assemblage composition, and benthic consumer assemblage structure, varied significantly with the intertidal influence of coastal upwelling, especially contrasting bays and coastal headlands. Coastal topography also separated differences in consumer resource use. This suggested that coastal upwelling, itself driven by coastline topography, influences intertidal communities by advecting nearshore phytoplankton populations offshore and cooling coastal water temperatures. We recommend the isotopic values of benthic organisms, specifically long-lived suspension feeders, as in situ alternatives to offshore measurements of upwelling influence. PMID:26214806

  18. SpBase: the sea urchin genome database and web site

    PubMed Central

    Cameron, R. Andrew; Samanta, Manoj; Yuan, Autumn; He, Dong; Davidson, Eric

    2009-01-01

    SpBase is a system of databases focused on the genomic information from sea urchins and related echinoderms. It is exposed to the public through a web site served with open source software (http://spbase.org/). The enterprise was undertaken to provide an easily used collection of information to directly support experimental work on these useful research models in cell and developmental biology. The information served from the databases emerges from the draft genomic sequence of the purple sea urchin, Strongylocentrotus purpuratus and includes sequence data and genomic resource descriptions for other members of the echinoderm clade which in total span 540 million years of evolutionary time. This version of the system contains two assemblies of the purple sea urchin genome, associated expressed sequences, gene annotations and accessory resources. Search mechanisms for the sequences and the gene annotations are provided. Because the system is maintained along with the Sea Urchin Genome resource, a database of sequenced clones is also provided. PMID:19010966

  19. Assessment of sediment toxicity and chemical concentrations in the San Diego Bay region, California, USA

    SciTech Connect

    Fairey, R.; Roberts, C.; Jacobi, M.

    1998-08-01

    Sediment quality within San Diego Bay, Mission Bay, and the Tijuana River Estuary of California was investigated as part of an ongoing statewide monitoring effort (Bay Protection and Toxic Cleanup Program). Study objectives were to determine the incidence, spatial patterns, and spatial extent of toxicity in sediments and porewater; the concentration and distribution of potentially toxic anthropogenic chemicals; and the relationships between toxicity and chemical concentrations. Rhepoxynius abronius survival bioassays, grain size, and total organic carbon analyses were performed on 350 sediment samples. Strongylocentrotus purpuratus development bioassays were performed on 164 pore-water samples. Toxicity was demonstrated throughout the San Diego Bay region, with increased incidence and concordance occurring in areas of industrial and shipping activity. Trace metal and trace synthetic organic analyses were performed on 229 samples. Copper, zinc, mercury, polycyclic aromatic hydrocarbons, polychlorinated biphenyls, and chlordane were found to exceed ERM (effects range median) or PEL (probable effects level) sediment quality guidelines and were considered the six major chemicals or chemical groups of concern. Statistical analysis of the relationships between amphipod toxicity, bulk phase sediment chemistry, and physical parameters demonstrated few significant linear relationships. Significant differences in chemical levels were found between toxic and nontoxic responses using multivariate and univariate statistics. Potential sources of anthropogenic chemicals were discussed.

  20. Kelp forest monitoring 1992 annual report. Channel Islands National Park. Final report

    SciTech Connect

    Richards, D.; Kushner, D.

    1992-12-31

    The 1992 results of the Channel Islands Natioanl Park Kelp Forest Monitoring Project are described in this report. Population dynamics of 68 taxa or categories of algae, fish, and invertebrates were measured at 16 permanent sites around the five islands within the park. Survey techniques utilized SCUBA and surface-supplied-air, and included quadrats, band transects, random point contacts, fish and video transects, photogrammetric plots, size frequency measurements, artifical recruitment habitats, and species list surveys. Some batheothermograph data was collected. In 1992, nine sites and healthy kelp forests while seven were mostly barren. The seven barren sites consisted of one that was dominated by the aggregated red sea cucumber, Pachythyone rubra, one was barren with high sedimentation, one was domainated by red sea urchins, Strongylocentrotus franciscanus, and four sites were dominated by purple sea urchins, Strongylocentrotus purpuratus, three of which had signs of a developing kelp forest. Wasting disease was observed in sea stars and a wasting syndrome was observed in sea urchins. Fish recruitment appeared to be late this year. Size frequency measurements were taken from artificial recruitment modules (previously named `abalone recruitment modules`) at six of the sites.

  1. High-Density Genetic Mapping with Interspecific Hybrids of Two Sea Urchins, Strongylocentrotus nudus and S. intermedius, by RAD Sequencing

    PubMed Central

    Dong, Ying; Gao, Shan; Chen, Zhong; Jiang, Jingwei; Yang, Aifu; Sun, Hongjuan; Guan, Xiaoyan; Jiang, Bei; Wang, Bai

    2015-01-01

    Sea urchins have long been used as research model organisms for developmental biology and evolutionary studies. Some of them are also important aquaculture species in East Asia. In this work, we report the construction of RAD-tag based high-density genetic maps by genotyping F1 interspecific hybrids derived from a crossing between a female sea urchin Strongylocentrotus nudus and a male Strongylocentrotus intermedius. With polymorphisms present in these two wild individuals, we constructed a female meiotic map containing 3,080 markers for S. nudus, and a male meiotic map for S. intermedius which contains 1,577 markers. Using the linkage maps, we were able to anchor a total of 1,591 scaffolds (495.9 Mb) accounting for 60.8% of the genome assembly of Strongylocentrotus purpuratus. A genome-wide scan resulted in the identification of one putative QTL for body size which spanned from 25.3 cM to 30.3 cM. This study showed the efficiency of RAD-Seq based high-density genetic map construction using F1 progenies for species with no prior genomic information. The genetic maps are essential for QTL mapping and are useful as framework to order and orientate contiguous scaffolds from sea urchin genome assembly. The integration of the genetic map with genome assembly would provide an unprecedented opportunity to conduct QTL analysis, comparative genomics, and population genetics studies. PMID:26398139

  2. [Evolutional principles of homology in regulatory genes of myogenesis].

    PubMed

    Ozerniuk, I D; Miuge, N S

    2012-01-01

    Analysis of early steps in muscular system development of invertebrates and vertebrates shows that early steps of myogenesis are regulated by genes-orthologs mainly belonging to two families, Pax and bHLH. In the majority of the following organisms, muscles formation (steps of determination and the earliest steps of myogenesis) is regulated by genes orthologs Pax3 which belong to the family Pax: nematodes (Caenorhabditis elegans, Pristionchus pacificus), insects (Drosophila melanogaster), echinoderms (Strongylocentrotus purpuratus), sea squirts (Ciona intestinalis, Holocynthia roretzi), fishes (Danio rerio), amphibians (Xenopus laevis), birds, and mammals (mouse, rat). The nematode C. elegans is an exception since formation of its muscles in this period is regulated by homeobox gene Pal-1 belonging to the family Caudal. The sea squirt C. intestinalis is also an exception because the earliest steps of development involved in further muscle formation are accompanied by activation of the gene CiSna (snail) (gene family basic Zinc finger). The next steps of myogenesis in all analyzed species are regulated by genes orthologs belonging to the family of transcriptional factors bHLH. They along with genes Pax3 are characterized by a high extent of homology in all studied groups of animals. PMID:22988754

  3. Extraordinary Diversity of Immune Response Proteins among Sea Urchins: Nickel-Isolated Sp185/333 Proteins Show Broad Variations in Size and Charge.

    PubMed

    Sherman, Lauren S; Schrankel, Catherine S; Brown, Kristy J; Smith, L Courtney

    2015-01-01

    Effective protection against pathogens requires the host to produce a wide range of immune effector proteins. The Sp185/333 gene family, which is expressed by the California purple sea urchin Strongylocentrotus purpuratus in response to bacterial infection, encodes a highly diverse repertoire of anti-pathogen proteins. A subset of these proteins can be isolated by affinity to metal ions based on multiple histidines, resulting in one to four bands of unique molecular weight on standard Western blots, which vary depending on the individual sea urchin. Two dimensional gel electrophoresis (2DE) of nickel-isolated protein samples followed by Western blot was employed to detect nickel-isolated Sp185/333 (Ni-Sp185/333) proteins and to evaluate protein diversity in animals before and after immune challenge with marine bacteria. Ni-Sp185/333 proteins of the same molecular weight on standard Western blots appear as a broad complex of variants that differ in pI on 2DE Western blots. The Ni-Sp185/333 protein repertoire is variable among animals, and shows a variety of changes among individual sea urchins in response to immune challenges with both the same and different species of bacteria. The extraordinary diversity of the Ni-Sp185/333 proteins may provide significant anti-pathogen capabilities for sea urchins that survive solely on innate immunity. PMID:26406912

  4. Identification and comparative analysis of complement C3-associated microRNAs in immune response of Apostichopus japonicus by high-throughput sequencing

    PubMed Central

    Zhong, Lei; Zhang, Feng; Zhai, Yu; Cao, Yanhui; Zhang, Si; Chang, Yaqing

    2015-01-01

    MicroRNAs (miRNAs) are important effectors in mediating host–pathogen interaction. In this report, coelomocytes miRNA libraries of three Japanese sea cucumbers Apostichopus japonicus were built by Illumina® Hiseq2000 from different time points after lipopolysaccharide challenge (at time 0 h, 6 h and 12 h). The clean data received from high throughput sequencing were used to sequences analysis. Referenced to the Strongylocentrotus purpuratus genome, 38 conserved miRNAs were found, and three miRNA candidates were predicted by software. According to the evidence resulting from the expression of AjC3, expressing levels of spu-miR-133, spu-miR-137 and spu-miR-2004 altered along with the expression of AjC3 changing at different time points after LPS injection. Thus, we speculated that the three miRNAs may have influence on A. japonicus complement C3. The spu-miR-137 and miR-137 gene family in miRBase were analyzed by bioinformatics. There is an obvious discrepancy between invertebrates and vertebrates. The first and ninth nucleotides in invertebrate miR-137 are offset compared vertebrate miR-137. Importantly, this is the first attempt to map the stage of immune response regulome in echinoderms, which might be considered as information for elucidating the intrinsic mechanism underlying the immune system in this species. PMID:26634300

  5. Extraordinary Diversity of Immune Response Proteins among Sea Urchins: Nickel-Isolated Sp185/333 Proteins Show Broad Variations in Size and Charge

    PubMed Central

    Sherman, Lauren S.; Schrankel, Catherine S.; Brown, Kristy J.; Smith, L. Courtney

    2015-01-01

    Effective protection against pathogens requires the host to produce a wide range of immune effector proteins. The Sp185/333 gene family, which is expressed by the California purple sea urchin Strongylocentrotus purpuratus in response to bacterial infection, encodes a highly diverse repertoire of anti-pathogen proteins. A subset of these proteins can be isolated by affinity to metal ions based on multiple histidines, resulting in one to four bands of unique molecular weight on standard Western blots, which vary depending on the individual sea urchin. Two dimensional gel electrophoresis (2DE) of nickel-isolated protein samples followed by Western blot was employed to detect nickel-isolated Sp185/333 (Ni-Sp185/333) proteins and to evaluate protein diversity in animals before and after immune challenge with marine bacteria. Ni-Sp185/333 proteins of the same molecular weight on standard Western blots appear as a broad complex of variants that differ in pI on 2DE Western blots. The Ni-Sp185/333 protein repertoire is variable among animals, and shows a variety of changes among individual sea urchins in response to immune challenges with both the same and different species of bacteria. The extraordinary diversity of the Ni-Sp185/333 proteins may provide significant anti-pathogen capabilities for sea urchins that survive solely on innate immunity. PMID:26406912

  6. Analysis of paralytic shellfish toxins and their metabolites in shellfish from the North Yellow Sea of China.

    PubMed

    Li, A; Ma, J; Cao, J; Wang, Q; Yu, R; Thomas, K; Quilliam, M A

    2012-01-01

    Samples of toxic scallop (Patinopecten yessoensis) and clam (Saxidomus purpuratus) collected on the northern coast of China from 2008 to 2009 were analysed. High-performance liquid chromatography with post-column oxidation and fluorescence detection was used to determine the profile of the main paralytic shellfish poisoning (PSP) toxins in these samples and their total toxicity. Hydrophilic interaction liquid ion chromatography with mass spectrometric detection confirmed the toxin profile and detected several metabolites in the shellfish. Results show that C1/2 toxins were the most dominant toxins in the scallop and clam samples. However, GTX1/4 and GTX2/3 were also present. M1 was the predominant metabolite in all the samples, but M3 and M5 were also identified, along with three previously unreported presumed metabolites, M6, M8 and M10. The results indicate that the biotransformation of toxins was species specific. It was concluded that the reductive enzyme in clams is more active than in scallops and that an enzyme in scallops is more apt to catalyse hydrolysis of both the sulfonate moiety at the N-sulfocabamoyl of C toxins and the 11-hydroxysulfate of C and GTX toxins to produce metabolites. This is the first report of new metabolites of PSP toxins in scallops and clams collected in China. PMID:22827221

  7. A Novel Fatty Acid-Binding Protein-Like Carotenoid-Binding Protein from the Gonad of the New Zealand Sea Urchin Evechinus chloroticus

    PubMed Central

    Pilbrow, Jodi; Sabherwal, Manya; Garama, Daniel; Carne, Alan

    2014-01-01

    A previously uncharacterized protein with a carotenoid-binding function has been isolated and characterized from the gonad of the New Zealand sea urchin Evechinus chloroticus. The main carotenoid bound to the protein was determined by reversed phase-high performance liquid chromatography to be 9′-cis-echinenone and hence this 15 kDa protein has been called an echinenone-binding protein (EBP). Purification of the EBP in quantity from the natural source proved to be challenging. However, analysis of EBP by mass spectrometry combined with information from the Strongylocentrotus purpuratus genome sequence and the recently published E. chloroticus transcriptome database, enabled recombinant expression of wild type EBP and also of a cysteine61 to serine mutant that had improved solubility characteristics. Circular dichroism data and ab initio structure prediction suggests that the EBP adopts a 10-stranded β-barrel fold consistent with that of fatty acid-binding proteins. Therefore, EBP may represent the first report of a fatty acid-binding protein in complex with a carotenoid. PMID:25192378

  8. A novel fatty acid-binding protein-like carotenoid-binding protein from the gonad of the New Zealand sea urchin Evechinus chloroticus.

    PubMed

    Pilbrow, Jodi; Sabherwal, Manya; Garama, Daniel; Carne, Alan

    2014-01-01

    A previously uncharacterized protein with a carotenoid-binding function has been isolated and characterized from the gonad of the New Zealand sea urchin Evechinus chloroticus. The main carotenoid bound to the protein was determined by reversed phase-high performance liquid chromatography to be 9'-cis-echinenone and hence this 15 kDa protein has been called an echinenone-binding protein (EBP). Purification of the EBP in quantity from the natural source proved to be challenging. However, analysis of EBP by mass spectrometry combined with information from the Strongylocentrotus purpuratus genome sequence and the recently published E. chloroticus transcriptome database, enabled recombinant expression of wild type EBP and also of a cysteine61 to serine mutant that had improved solubility characteristics. Circular dichroism data and ab initio structure prediction suggests that the EBP adopts a 10-stranded β-barrel fold consistent with that of fatty acid-binding proteins. Therefore, EBP may represent the first report of a fatty acid-binding protein in complex with a carotenoid. PMID:25192378

  9. The Sp185/333 immune response genes and proteins are expressed in cells dispersed within all major organs of the adult purple sea urchin.

    PubMed

    Majeske, Audrey J; Oleksyk, Taras K; Smith, L Courtney

    2013-12-01

    Purple sea urchins (Strongylocentrotus purpuratus) express a highly variable set of immune genes called Sp185/333 by two subtypes of coelomocytes: the polygonal and small phagocytes. We report that the Sp185/333 genes and their encoded proteins are also expressed in all of the major organs in the adult sea urchin, including the axial organ, pharynx, esophagus, intestine and gonads. After immune challenge, there is an increase in the level of Sp185/333 mRNA in cells associated with the intestine and axial organ. The Sp185/333 proteins increase in the axial organ, pharynx, esophagus and intestine after challenge. However, the proportion of Sp185/333-positive cells only increases in the axial organ, while there is no change in that proportion in the other organs after challenge. The size range of the major Sp185/333 proteins expressed by organs is broader (5 kDa to > 250 kDa) compared with those in coelomocytes (∼40 kDa to < 250 kDa). Images of the different organs do not clarify whether coelomocytes or parenchymal cells express the Sp185/333 proteins. The increase in levels of Sp185/333 transcripts, protein expression and Sp185/333-positive cells in the axial organ in response to challenge suggests that this organ may have an important role in immunity for this species. PMID:23405032

  10. Marine invertebrate genome sequences and our evolving understanding of animal immunity.

    PubMed

    Rast, Jonathan P; Messier-Solek, Cynthia

    2008-06-01

    Animal immunity is under intense evolutionary pressure, and the mechanisms that carry out recognition and elimination of pathogens are among the most rapidly evolving genetic systems. It is increasingly apparent that this has led to the emergence of novel molecular mechanisms not only among vertebrates, where immunity is by far best characterized, but also across invertebrate phyla. This propensity for rapid divergence has been a serious obstacle for progress in the field of comparative immunology. The variety of recent genome sequences from marine invertebrates representing new phyla offers a means to move forward in this area. Genome sequences provide much improved sensitivity for the detection of gene homologs and a framework for unbiased computational and experimental searches for novel immune mediators. Furthermore, new genomes now offer a more complete and unbiased view of immunity across bilaterian phyla, especially among deuterostomes. In this review we summarize these findings with particular attention toward immunity in Strongylocentrotus purpuratus, the purple sea urchin, and outline the changing perspective on the evolution of deuterostome immunity. PMID:18574104

  11. Genome-wide polymorphisms show unexpected targets of natural selection.

    PubMed

    Pespeni, Melissa H; Garfield, David A; Manier, Mollie K; Palumbi, Stephen R

    2012-04-01

    Natural selection can act on all the expressed genes of an individual, leaving signatures of genetic differentiation or diversity at many loci across the genome. New power to assay these genome-wide effects of selection comes from associating multi-locus patterns of polymorphism with gene expression and function. Here, we performed one of the first genome-wide surveys in a marine species, comparing purple sea urchins, Strongylocentrotus purpuratus, from two distant locations along the species' wide latitudinal range. We examined 9112 polymorphic loci from upstream non-coding and coding regions of genes for signatures of selection with respect to gene function and tissue- and ontogenetic gene expression. We found that genetic differentiation (F(ST)) varied significantly across functional gene classes. The strongest enrichment occurred in the upstream regions of E3 ligase genes, enzymes known to regulate protein abundance during development and environmental stress. We found enrichment for high heterozygosity in genes directly involved in immune response, particularly NALP genes, which mediate pro-inflammatory signals during bacterial infection. We also found higher heterozygosity in immune genes in the southern population, where disease incidence and pathogen diversity are greater. Similar to the major histocompatibility complex in mammals, balancing selection may enhance genetic diversity in the innate immune system genes of this invertebrate. Overall, our results show that how genome-wide polymorphism data coupled with growing databases on gene function and expression can combine to detect otherwise hidden signals of selection in natural populations. PMID:21993504

  12. Toxicity of urban highway runoff with respect to storm duration.

    PubMed

    Kayhanian, M; Stransky, C; Bay, S; Lau, S-L; Stenstrom, M K

    2008-01-25

    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. PMID:17920106

  13. The Drosophila erect wing gene, which is important for both neuronal and muscle development, encodes a protein which is similar to the sea urchin P3A2 DNA binding protein.

    PubMed Central

    DeSimone, S M; White, K

    1993-01-01

    The erect wing (ewg) locus of Drosophila melanogaster encodes a vital function important for the development of the nervous system and the indirect flight muscles. In order to understand the ewg function at a molecular level, cDNA clones were isolated. Sequence analysis of cDNAs revealed a single open reading frame (ORF) encoding a protein of 733 residues. The translational start for this ORF is a CTG codon. A 225-amino-acid region of this protein is 71% identical to the DNA binding region of the Strongylocentrotus purpuratus P3A2 DNA binding protein. Additionally, the ORF contains large acidic and basic domains characteristic of those in proteins involved in nuclear regulatory functions. Immunoblot analysis using polyclonal anti-EWG antisera generated against a bacterial fusion protein reveals a single, 116-kDa protein present throughout development, beginning at approximately stage 12 of embryogenesis, which is enriched in adult heads and absent from embryos carrying certain ewg alleles. Additionally, we show that EWG is localized specifically to the nuclei of virtually all embryonic neurons. Finally, a minigene consisting of an ewg cDNA under control of the hsp70 promoter can provide the ewg function in transgenic ewg mutant flies. Images PMID:8388540

  14. Intermittent hypoxia leads to functional reorganization of mitochondria and affects cellular bioenergetics in marine molluscs.

    PubMed

    Ivanina, Anna V; Nesmelova, Irina; Leamy, Larry; Sokolov, Eugene P; Sokolova, Inna M

    2016-06-01

    Fluctuations in oxygen (O2) concentrations represent a major challenge to aerobic organisms and can be extremely damaging to their mitochondria. Marine intertidal molluscs are well-adapted to frequent O2 fluctuations, yet it remains unknown how their mitochondrial functions are regulated to sustain energy metabolism and prevent cellular damage during hypoxia and reoxygenation (H/R). We used metabolic control analysis to investigate the mechanisms of mitochondrial responses to H/R stress (18 h at <0.1% O2 followed by 1 h of reoxygenation) using hypoxia-tolerant intertidal clams Mercenaria mercenaria and hypoxia-sensitive subtidal scallops Argopecten irradians as models. We also assessed H/R-induced changes in cellular energy balance, oxidative damage and unfolded protein response to determine the potential links between mitochondrial dysfunction and cellular injury. Mitochondrial responses to H/R in scallops strongly resembled those in other hypoxia-sensitive organisms. Exposure to hypoxia followed by reoxygenation led to a strong decrease in the substrate oxidation (SOX) and phosphorylation (PHOS) capacities as well as partial depolarization of mitochondria of scallops. Elevated mRNA expression of a reactive oxygen species-sensitive enzyme aconitase and Lon protease (responsible for degradation of oxidized mitochondrial proteins) during H/R stress was consistent with elevated levels of oxidative stress in mitochondria of scallops. In hypoxia-tolerant clams, mitochondrial SOX capacity was enhanced during hypoxia and continued rising during the first hour of reoxygenation. In both species, the mitochondrial PHOS capacity was suppressed during hypoxia, likely to prevent ATP wastage by the reverse action of FO,F1-ATPase. The PHOS capacity recovered after 1 h of reoxygenation in clams but not in scallops. Compared with scallops, clams showed a greater suppression of energy-consuming processes (such as protein turnover and ion transport) during hypoxia, indicated

  15. Magmatic evolution of pre-ore volcanics and porphyry intrusives associated with the Altar Cu-porphyry prospect, Argentina

    NASA Astrophysics Data System (ADS)

    Gatzoubaros, M.; von Quadt, A.; Gallhofer, D.; Rey, R.

    2014-11-01

    Altar is a Cu-porphyry deposit related to several small plagioclase porphyry intrusions of the late Miocene formed on the margin of the Flat-Slab segment along the Andean Cordillera in north-west Argentina. New stratigraphic and structural mapping supported by geochemistry and geochronology of pre-ore volcanics at Altar has revealed that a period of ˜6-7 Ma of volcanism during the late Oligocene-early Miocene formed ˜4000 m of volcano-stratigraphic succession making up the Pachón Formation. It represents a period dominated by explosive to effusive eruption in a dynamic arc basin with local ash fall and flow deposition in lacustrine and fluvial sites. Volcanism is typified by medium- to high-K calc-alkaline arc magmatism with a shift from mafic compositions at the base to felsic rocks at the top of the formation containing zircons aged 21.9 ± 0.2 Ma (2 Std.Dev, U-Pb). A clear geochemical separation exists between early Miocene pre-ore volcanics that show signatures akin to non-adakitic, normal arc, extensional tectonic settings conducive of chemical differentiation at shallow crustal levels and correlate with intra-regional Abanico and Farellones Formations; and the middle to late-Miocene Cu-mineralised porphyry intrusions. After a break of ˜9 Ma in the geological record at Altar, these Cu-fertile bodies are emplaced entirely within the Pachón Rhyolite and represent adakite-like magmas with fractionation trends evolving from a lower crustal MASH zone. This distinction is controlled by a change from an extensional to compressive tectonic regime in the region during the middle Miocene in which magmas were stalled in the lower crust for an extended period, subsequently became enriched in metals and then formed several Cu-porphyry bodies which were emplaced during a relatively short period towards the late Miocene.

  16. Construction of an Upper Crustal Reservoir by Lateral Magma Propagation: New insights from Geochronological Data of La Gloria Pluton, Central Chile

    NASA Astrophysics Data System (ADS)

    Gutiérrez, F. J.; Guillong, M.; Payacán, I. J.; Aravena, A.; Bachmann, O.; Parada, M. A.

    2014-12-01

    La Gloria Pluton (LGP) is a 10 Ma shallow elongated NNW reservoir of 17 km length and 4-6 km width as part of a NS trend of Miocene plutons in Central Chile. New LA-ICPMS U-Pb ages in zircons of La Gloria Pluton indicate that crystallization occurs mostly within an interval between 11.2 to 10 Ma, with southeastward decreasing ages. Zircon crystallization ages are consistently older at the boundaries of the pluton than at the center for a given cross-section. At regional scale the ages of LGP follows a plutonic trend of southward decreasing age: Estero Yerba Loca (10 Ma) and San Francisco Batholith (SFB), in the north; and Cerro Mesón Alto (12.5 Ma) and San Gabriel (SG; 13 Ma), in the south. Both regional and local (within-LGP) age trends suggest: 1) a progressive northward migration of the main deep magmatic source during the Miocene; and 2) a southeastward lateral propagation of the magma during the reservoir construction. The lateral propagation of the magma is also supported by subhorizontal mineral and magnetic lineations with a preferred NNW orientation within LGP. The within-pluton age distribution and internal configuration suggest incremental construction with horizontal propagation of magma within channels. Because the lateral migration of the magma play an important role on the thermal structure of the cooling pluton we perform numerical simulations that account for reheating caused by refilling along the axial core of the pluton . We speculate that pre-existing shallow crustal structures (faults and folds) would allow lateral magma canalization, particularly between the lower highly deformed volcanic Abanico Fm. and the less deformed overlaying volcanic Farellones Fm. The pluton distribution and internal organization in and around LGP suggest incremental construction with vertical and horizontal migration of magma within channels and reservoirs, yielding plutonic complexes with protracted ages and elongated geometries.

  17. Chronologic implications of new Miocene mammals from the Cura-Mallín and Trapa Trapa formations, Laguna del Laja area, south central Chile

    NASA Astrophysics Data System (ADS)

    Flynn, John J.; Charrier, Reynaldo; Croft, Darin A.; Gans, Phillip B.; Herriott, Trystan M.; Wertheim, Jill A.; Wyss, André R.

    2008-12-01

    Recent work in the central Andean Main Range of Chile near Laguna del Laja (˜37.5°S, 71°W) has produced the first mammal fossils for the region. Fossils, locally abundant and well preserved, occur patchily across a wide area southeast of the lake. Mammalian remains are derived from generally strongly folded (kilometer-scale) exposures of the locally ˜1.8 km thick, early to middle Miocene Cura-Mallín Formation; two identifiable specimens have been recovered from the overlying Trapa Trapa Formation as well. Both formations consist primarily of well-stratified (1-5 m thick layers) volcaniclastic and volcanic strata, deposited predominantly in fluviatile systems. The Cura-Mallín Formation is possibly the southern continuation of (or lateral equivalent to) the richly fossiliferous Abanico Formation mapped between ˜32°S and 36°S. Intensive sampling in a series of localities east and south of Laguna del Laja has yielded diverse faunas, in addition to radioisotopically dateable horizons. The new fossil mammal faunas represent as many as six South American Land Mammal "Ages" (SALMAs). Fossils, together with preliminary 40Ar/ 39Ar radioisotopic dates, ranging from ˜9 to 20 Ma across the exposed thickness of the Cura-Mallín Formation and into the overlying Trapa Trapa Formation, provide a robust geochronological framework for middle Cenozoic strata in the Laguna del Laja region. The sequence of directly superposed mammalian assemblages at Laguna del Laja is one of the longest in all of South America, rivaled only by the classic Gran Barranca section of Patagonian Argentina. These data illuminate the geological history of the area and its record of mammalian evolution. The potential to isotopically date these diverse faunas with high precision (error ± 0.5 Ma) presents a rare opportunity to calibrate related portions of the SALMA sequence.

  18. A putative 'pre-nervous' endocannabinoid system in early echinoderm development.

    PubMed

    Buznikov, G A; Nikitina, L A; Bezuglov, V V; Francisco, M E Y; Boysen, G; Obispo-Peak, I N; Peterson, R E; Weiss, E R; Schuel, H; Temple, B R S; Morrow, A L; Lauder, J M

    2010-03-01

    Embryos and larvae of sea urchins (Lytechinus variegatus, Strongylocentrotus droebachiensis, Strongylocentrotus purpuratus, Dendraster excentricus), and starfish (Pisaster ochraceus) were investigated for the presence of a functional endocannabinoid system. Anandamide (arachidonoyl ethanolamide, AEA), was measured in early L. variegatus embryos by liquid chromatography/mass spectrometry. AEA showed a strong developmental dynamic, increasing more than 5-fold between the 8-16 cell and mid-blastula 2 stage. 'Perturb-and-rescue' experiments in different sea urchin species and starfish showed that AEA blocked transition of embryos from the blastula to the gastrula stage, but had no effect on cleavage divisions, even at high doses. The non-selective cannabinoid receptor agonist, CP55940, had similar effects, but unlike AEA, also blocked cleavage divisions. CB1 antagonists, AEA transport inhibitors, and the cation channel transient membrane potential receptor V1 (TrpV1) agonist, arachidonoyl vanillic acid (arvanil), as well as arachidonoyl serotonin and dopamine (AA-5-HT, AA-DA) acted as rescue substances, partially or totally preventing abnormal embryonic phenotypes elicited by AEA or CP55940. Radioligand binding of [(3)H]CP55940 to membrane preparations from embryos/larvae failed to show significant binding, consistent with the lack of CB receptor orthologs in the sea urchin genome. However, when binding was conducted on whole cell lysates, a small amount of [(3)H]CP55940 binding was observed at the pluteus stage that was displaced by the CB2 antagonist, SR144528. Since AEA is known to bind with high affinity to TrpV1 and to certain G-protein-coupled receptors (GPCRs), the ability of arvanil, AA-5-HT and AA-DA to rescue embryos from AEA teratogenesis suggests that in sea urchins AEA and other endocannabinoids may utilize both Trp and GPCR orthologs. This possibility was explored using bioinformatic and phylogenetic tools to identify candidate orthologs in the S

  19. A Genomic View of the Sea Urchin Nervous System

    PubMed Central

    Burke, RD; Angerer, LM; Elphick, MR; Humphrey, GW; Yaguchi, S; Kiyama, T; Liang, S; Mu, X; Agca, C; Klein, WH; Brandhorst, BP; Rowe, M; Wilson, K; Churcher, AM; Taylor, JS; Chen, N; Murray, G; Wang, D; Mellott, D; Olinski, R; Hallböök, F; Thorndyke, MC

    2007-01-01

    The sequencing of the Strongylocentrotus purpuratus genome provides a unique opportunity to investigate the function and evolution of neural genes. The neurobiology of sea urchins is of particular interest because they have a close phylogenetic relationship with chordates, yet a distinctive pentaradiate body plan and unusual neural organization. Orthologues of transcription factors that regulate neurogenesis in other animals have been identified and several are expressed in neurogenic domains before gastrulation indicating that they may operate near the top of a conserved neural gene regulatory network. A family of genes encoding voltage-gated ion channels is present but, surprisingly, genes encoding gap junction proteins (connexins and pannexins) appear to be absent. Genes required for synapse formation and function have been identified and genes for synthesis and transport of neurotransmitters are present. There is a large family of G-protein-coupled receptors, including 874 rhodopsin-type receptors, 28 metabotropic glutamate-like receptors and a remarkably expanded group of 161 secretin receptor-like proteins. Absence of cannabinoid, lysophospholipid and melanocortin receptors indicates that this group may be unique to chordates. There are at least 37 putative G-protein coupled peptide receptors and precursors for several neuropeptides and peptide hormones have been identified, including SALMFamides, NGFFFamide, a vasotocin-like peptide, glycoprotein hormones, and insulin/insulin-like growth factors. Identification of a neurotrophin-like gene and Trk receptor in sea urchin indicates that this neural signaling system is not unique to chordates. Several hundred chemoreceptor genes have been predicted using several approaches, a number similar to that for other animals. Intriguingly, genes encoding homologues of rhodopsin, Pax6 and several other key mammalian retinal transcription factors are expressed in tube feet, suggesting tube feet function as photosensory

  20. Turbulent shear spurs settlement in larval sea urchins

    PubMed Central

    Gaylord, Brian; Hodin, Jason; Ferner, Matthew C.

    2013-01-01

    Marine invertebrates commonly produce larvae that disperse in ocean waters before settling into adult shoreline habitat. Chemical and other seafloor-associated cues often facilitate this latter transition. However, the range of effectiveness of such cues is limited to small spatial scales, creating challenges for larvae in finding suitable sites at which to settle, especially given that they may be carried many kilometers by currents during their planktonic phase. One possible solution is for larvae to use additional, broader-scale environmental signposts to first narrow their search to the general vicinity of a candidate settlement location. Here we demonstrate strong effects of just such a habitat-scale cue, one with the potential to signal larvae that they have arrived in appropriate coastal areas. Larvae of the purple sea urchin (Strongylocentrotus purpuratus) exhibit dramatic enhancement in settlement following stimulation by turbulent shear typical of wave-swept shores where adults of this species live. This response manifests in an unprecedented fashion relative to previously identified cues. Turbulent shear does not boost settlement by itself. Instead, it drives a marked developmental acceleration that causes “precompetent” larvae refractory to chemical settlement inducers to immediately become “competent” and thereby reactive to such inducers. These findings reveal an unrecognized ability of larval invertebrates to shift the trajectory of a major life history event in response to fluid-dynamic attributes of a target environment. Such an ability may improve performance and survival in marine organisms by encouraging completion of their life cycle in advantageous locations. PMID:23572585

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

    PubMed

    Fugmann, Sebastian D; Messier, Cynthia; Novack, Laura A; Cameron, R Andrew; Rast, Jonathan P

    2006-03-01

    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. PMID:16505374

  2. Transcriptome Sequencing and Characterization for the Sea Cucumber Apostichopus japonicus (Selenka, 1867)

    PubMed Central

    Du, Huixia; Bao, Zhenmin; Hou, Rui; Wang, Shan; Su, Hailin; Yan, Jingjing; Tian, Meilin; Li, Yan; Wei, Wen; Lu, Wei; Hu, Xiaoli; Wang, Shi; Hu, Jingjie

    2012-01-01

    Background Sea cucumbers are a special group of marine invertebrates. They occupy a taxonomic position that is believed to be important for understanding the origin and evolution of deuterostomes. Some of them such as Apostichopus japonicus represent commercially important aquaculture species in Asian countries. Many efforts have been devoted to increasing the number of expressed sequence tags (ESTs) for A. japonicus, but a comprehensive characterization of its transcriptome remains lacking. Here, we performed the large-scale transcriptome profiling and characterization by pyrosequencing diverse cDNA libraries from A. japonicus. Results In total, 1,061,078 reads were obtained by 454 sequencing of eight cDNA libraries representing different developmental stages and adult tissues in A. japonicus. These reads were assembled into 29,666 isotigs, which were further clustered into 21,071 isogroups. Nearly 40% of the isogroups showed significant matches to known proteins based on sequence similarity. Gene ontology (GO) and KEGG pathway analyses recovered diverse biological functions and processes. Candidate genes that were potentially involved in aestivation were identified. Transcriptome comparison with the sea urchin Strongylocentrotus purpuratus revealed similar patterns of GO term representation. In addition, 4,882 putative orthologous genes were identified, of which 202 were not present in the non-echinoderm organisms. More than 700 simple sequence repeats (SSRs) and 54,000 single nucleotide polymorphisms (SNPs) were detected in the A. japonicus transcriptome. Conclusion Pyrosequencing was proven to be efficient in rapidly identifying a large set of genes for the sea cucumber A. japonicus. Through the large-scale transcriptome sequencing as well as public EST data integration, we performed a comprehensive characterization of the A. japonicus transcriptome and identified candidate aestivation-related genes. A large number of potential genetic markers were also

  3. Unique system of photoreceptors in sea urchin tube feet

    PubMed Central

    Ullrich-Lüter, Esther M; Dupont, Sam; Arboleda, Enrique; Hausen, Harald; Arnone, Maria Ina

    2011-01-01

    Different sea urchin species show a vast variety of responses to variations in light intensity; however, despite this behavioral evidence for photosensitivity, light sensing in these animals has remained an enigma. Genome information of the recently sequenced purple sea urchin (Strongylocentrotus purpuratus) allowed us to address this question from a previously unexplored molecular perspective by localizing expression of the rhabdomeric opsin Sp-opsin4 and Sp-pax6, two genes essential for photoreceptor function and development, respectively. Using a specifically designed antibody against Sp-Opsin4 and in situ hybridization for both genes, we detected expression in two distinct groups of photoreceptor cells (PRCs) located in the animal's numerous tube feet. Specific reactivity of the Sp-Opsin4 antibody with sea star optic cushions, which regulate phototaxis, suggests a similar visual function in sea urchins. Ultrastructural characterization of the sea urchin PRCs revealed them to be of a microvillar receptor type. Our data suggest that echinoderms, in contrast to chordates, deploy a microvillar, r-opsin–expressing PRC type for vision, a feature that has been so far documented only in protostome animals. Surprisingly, sea urchin PRCs lack any associated screening pigment. Indeed, one of the tube foot PRC clusters may account for directional vision by being shaded through the opaque calcite skeleton. The PRC axons connect to the animal internal nervous system, suggesting an integrative function beyond local short circuits. Because juveniles display no phototaxis until skeleton completion, we suggest a model in which the entire sea urchin, deploying its skeleton as PRC screening device, functions as a huge compound eye. PMID:21536888

  4. A Model Sea Urchin Spicule Matrix Protein Self-Associates To Form Mineral-Modifying Protein Hydrogels.

    PubMed

    Jain, Gaurav; Pendola, Martin; Rao, Ashit; Cölfen, Helmut; Evans, John Spencer

    2016-08-01

    In the purple sea urchin Strongylocentrotus purpuratus, the formation and mineralization of fracture-resistant skeletal elements such as the embryonic spicule require the combinatorial participation of numerous spicule matrix proteins such as the SpSM30A-F isoforms. However, because of limited abundance, it has been difficult to pursue extensive biochemical studies of the SpSM30 proteins and deduce their role in spicule formation and mineralization. To circumvent these problems, we expressed a model recombinant spicule matrix protein, rSpSM30B/C, which possesses the key sequence attributes of isoforms "B" and "C". Our findings indicate that rSpSM30B/C is expressed in insect cells as a single polypeptide containing variations in glycosylation that create microheterogeneity in rSpSM30B/C molecular masses. These post-translational modifications incorporate O- and N-glycans and anionic mono- and bisialylated and mono- and bisulfated monosaccharides on the protein molecules and enhance its aggregation propensity. Bioinformatics and biophysical experiments confirm that rSpSM30B/C is an intrinsically disordered, aggregation-prone protein that forms porous protein hydrogels that control the in vitro mineralization process in three ways: (1) increase the time interval for prenucleation cluster formation and transiently stabilize an ACC polymorph, (2) promote and organize single-crystal calcite nanoparticles, and (3) promote faceted growth and create surface texturing of calcite crystals. These features are also common to mollusk shell nacre proteins, and we conclude that rSpSM30B/C is a spiculogenesis protein that exhibits traits found in other calcium carbonate mineral modification proteins. PMID:27426695

  5. Transformation and crystallization energetics of synthetic and biogenic amorphous calcium carbonate

    PubMed Central

    Radha, A. V.; Forbes, Tori Z.; Killian, Christopher E.; Gilbert, P. U. P. A.; Navrotsky, Alexandra

    2010-01-01

    Amorphous calcium carbonate (ACC) is a metastable phase often observed during low temperature inorganic synthesis and biomineralization. ACC transforms with aging or heating into a less hydrated form, and with time crystallizes to calcite or aragonite. The energetics of transformation and crystallization of synthetic and biogenic (extracted from California purple sea urchin larval spicules, Strongylocentrotus purpuratus) ACC were studied using isothermal acid solution calorimetry and differential scanning calorimetry. Transformation and crystallization of ACC can follow an energetically downhill sequence: more metastable hydrated ACC → less metastable hydrated ACC⇒anhydrous ACC ∼ biogenic anhydrous ACC⇒vaterite → aragonite → calcite. In a given reaction sequence, not all these phases need to occur. The transformations involve a series of ordering, dehydration, and crystallization processes, each lowering the enthalpy (and free energy) of the system, with crystallization of the dehydrated amorphous material lowering the enthalpy the most. ACC is much more metastable with respect to calcite than the crystalline polymorphs vaterite or aragonite. The anhydrous ACC is less metastable than the hydrated, implying that the structural reorganization during dehydration is exothermic and irreversible. Dehydrated synthetic and anhydrous biogenic ACC are similar in enthalpy. The transformation sequence observed in biomineralization could be mainly energetically driven; the first phase deposited is hydrated ACC, which then converts to anhydrous ACC, and finally crystallizes to calcite. The initial formation of ACC may be a first step in the precipitation of calcite under a wide variety of conditions, including geological CO2 sequestration. PMID:20810918

  6. The sea urchin metallothionein system: Comparative evaluation of the SpMTA and SpMTB metal-binding preferences☆

    PubMed Central

    Tomas, Mireia; Domènech, Jordi; Capdevila, Mercè; Bofill, Roger; Atrian, Sílvia

    2013-01-01

    Metallothioneins (MTs) constitute a superfamily of ubiquitous metal-binding proteins of low molecular weight and high Cys content. They are involved in metal homeostasis and detoxification, amongst other proposed biological functions. Two MT isoforms (SpMTA and SpMTB) have been reported in the echinoderm Strongylocentrotus purpuratus (sea urchin), both containing 20 Cys residues and presenting extremely similar sequences, although showing distinct tissular and ontogenic expression patterns. Although exhaustive information is available for the Cd(II)-SpMTA complex, this including the full resolution of its 3D structure, no data has been reported concerning either SpMTA Zn(II) and Cu(I) binding properties, or the characterization of SpMTB at protein level. In this work, both the SpMTA and SpMTB isoforms, as well as their separate α and β domains, have been recombinantly synthesized in the presence of Zn(II), Cd(II) or Cu(II), and the corresponding metal complexes have been analyzed using electrospray mass spectrometry, and CD, ICP-AES and UV–vis spectroscopies. The results clearly show a better performance of isoform A when binding Zn(II) and Cd(II), and of isoform B when coordinating Cu(I). Thus, our results confirm the differential metal binding preference of SpMTA and SpMTB, which, together with the reported induction pattern of the respective genes, highlights how also in Echinodermata the MT polymorphism may be linked to the evolution of different physiological roles. PMID:23847757

  7. Ammonia and sediment toxicity

    SciTech Connect

    Ogle, R.S.; Hansen, S.R.

    1994-12-31

    Ammonia toxicity to aquatic organisms has received considerable study, with most of these studies focusing on water column organisms. However, with the development and implementation of sediment (and pore water) toxicity tests, the toxicity of ammonia to benthic infauna and other sediment toxicity test organisms has become important, especially since sediment/porewater ammonia occurs at higher concentrations than in the water column. Unfortunately, there has been very little of this type information, especially for marine/estuarine organisms. This laboratory determined the toxicity of ammonia to three key marine/estuarine test organisms: the amphipod Eohaustorius estuarius, the bivalve Mytilus edulis, and the echinoderm Strongylocentrotus purpuratus. Because sediment/porewater pH can differ substantially from typical seawater pH, the toxicity evaluations covered a range of pH levels (6, 7, 8, and 9). Eohaustorius results indicate that while Total Ammonia increased in toxicity (measured as EC50) as pH increased (from 460 mg/L at pH 6, to 13 mg/L at pH 9), unionized ammonia toxicity decreased from 0.13 mg/L at pH 6 to 2.8 mg/L at pH 9. The amphipod was much less sensitive to ammonia than were the bivalve and echinoderm, with an unionized ammonia EC50 at pH 8 of 2.14 mg/L relative to 0.43 mg/L for the mussel and 0.13 mg/L for the purple urchin. These results are discussed with respect to design and interpretation of sediment toxicity test results, including an interpretation approach based on partitioning of Toxic Units (TU).

  8. Molecular characterization and expression profile of MAP2K1ip1/MP1 gene from tiger shrimp, Penaeus monodon.

    PubMed

    Yang, Lishi; Liu, Xianjun; Huang, Jianhua; Yang, Qibin; Qiu, Lihua; Liu, Wenjing; Jiang, Shigui

    2012-05-01

    MAPK kinase 1 interacting protein 1 (MAP2K1ip1) is an important scaffold proteins of the mitogen-activated protein kinase (MAPK) pathway that form an active signaling module and enhance the specificity and spatiality of MAPK signaling. In the present study, we identified and characterized a MAP2K1ip1 cDNA from tiger shrimp Penaeus monodon (designated as PmMAP2K1ip1). The open reading frame of PmMAP2K1ip1 is 372 bp encoding 123 amino-acid residues with a MAPK interaction domain. The predicted PmMAP2Kip1 protein is 13.6 KDa with the theoretical isoelectric point of 6.3. PmMAP2K1ip1 shared the highest amino acid with Nasonia vitripennis and Strongylocentrotus purpuratus, at 48% and 47.5%, respectively. Phylogenic analysis shows PmMAP2Kip1 is clustering with SpMAP2Kip1, and close to the group of MAP2Kip1s from insect. Furthermore, semiquantitative RT-PCR revealed PmMAP2Kip1 is widely distributed in most examined tissues except nerve, and high expressed in ovary, hemocyte, intestines and hepatopancreas. Meanwhile, PmMAP2k1ip1 is expressed ubiquitously during larval and sex gland development, and keep a high level at the initial development stage. Quantitative real time RT-PCR revealed PmMAP2K1ip1 were up-regulated by lipopolysaccharide and peptidoglycan (PGN) in haemocyte. These data reveal MAP2K1ip1 is a multifunction protein that involved development and immune response. It is benefit to characterize other MAPK signal genes and elucidate the molecular regulation mechanism of MAPK signaling in tiger shrimp. PMID:22209950

  9. Cathepsin L of the sea cucumber Apostichopus japonicus-molecular characterization and transcriptional response to Vibrio splendidus infection.

    PubMed

    Yang, Jingwen; Liu, Huihui; Zheng, Gang; Xiang, Xiaowei; Lv, Zhenming; Wang, Tianming

    2016-02-01

    Cathepsin L, a lysosomal endopeptidase, has been noted for its involvement in the innate immune response in invertebrates. Here, the cathepsin L cDNA of the sea cucumber Apostichopus japonicus (AjCatL) is identified from an EST library and then cloned by the rapid amplification of the cDNA ends (RACE) PCR. The full-length cDNA is 1678 bp long containing an open reading frame (ORF) of 1002 bp, an 80 bp 5' UTR and a 599 bp 3' UTR. The cDNA encodes 333 amino acid residues with a predicted molecular mass of 37.07 kDa and a theoretical isoelectric point (pI) of 5.01. The full-length AjCatL contains three active sites of eukaryotic thiol (cysteine) protease at positions 133-144, 278-288 and 295-314. Analysis of the predicted tertiary structure of prepro-CatL (17-333 aa) and mature-CatL (116-333 aa) reveals that the propeptide region (17-115 aa) blocks access to the substrate-binding cleft. Phylogenetic analysis shows that the AjCatL is clustered together with two other CatLs from Strongylocentrotus purpuratus. The enzymatic activity of AjCatL was verified using a substrate hydrolyzing assay with recombinant mAjCatL. Further analysis of real time-PCR demonstrates that the expression of AjCatL mRNA is significantly up-regulated in the coelomocytes in cases of infection with the common bacterial pathogen, Vibrio splendidus. This suggests that the AjCatL is likely to be involved in the immune response. PMID:26777896

  10. Molecular cloning of heat shock protein 10 (Hsp10) and 60 (Hsp60) cDNAs and their expression analysis under thermal stress in the sea cucumber Apostichopus japonicus.

    PubMed

    Xu, Dongxue; Sun, Lina; Liu, Shilin; Zhang, Libin; Ru, Xiaoshang; Zhao, Ye; Yang, Hongsheng

    2014-05-01

    Hsp10 and Hsp60 are important heat shock proteins (HSPs), which might be indispensable in the heat shock response and many other physiological processes. We obtained full-length cDNAs of genes hsp10 and hsp60 and classified their mRNA expression levels under thermal stress in the sea cucumber Apostichopus japonicus. Full-length hsp10 cDNA was 1528 bp containing a 5' untranslated region (UTR) of 83 bp, a 3' UTR of 1133 bp and an open reading frame (ORF) of 312 bp encoding 103 amino acid residues. Full-length hsp60 cDNA was 2560 bp containing a 118-bp 5' UTR, a 678-bp 3' UTR and a 1764-bp ORF encoding 586 amino acid residues. The deduced amino acid sequences of Hsp10 and Hsp60 shared the highest identity with sequences of the sea urchin Strongylocentrotus purpuratus, and phylogenetic trees showed that the evolution of Hsp10 and Hsp60 was almost in accord with the evolution of species. Further analysis by real-time PCR showed that the expression of hsp10 and hsp60 mRNA was highly up-regulated at 26 °C compared with other three groups (20, 22, 24 °C), and their expression in the intestine was in a time-dependent manner at 26 °C. The results suggested that hsp10 and hsp60 were involved in the heat-shock response in the sea cucumber A. japonicus. PMID:24721556

  11. Experimental ocean acidification alters the allocation of metabolic energy.

    PubMed

    Pan, T-C Francis; Applebaum, Scott L; Manahan, Donal T

    2015-04-14

    Energy is required to maintain physiological homeostasis in response to environmental change. Although responses to environmental stressors frequently are assumed to involve high metabolic costs, the biochemical bases of actual energy demands are rarely quantified. We studied the impact of a near-future scenario of ocean acidification [800 µatm partial pressure of CO2 (pCO2)] during the development and growth of an important model organism in developmental and environmental biology, the sea urchin Strongylocentrotus purpuratus. Size, metabolic rate, biochemical content, and gene expression were not different in larvae growing under control and seawater acidification treatments. Measurements limited to those levels of biological analysis did not reveal the biochemical mechanisms of response to ocean acidification that occurred at the cellular level. In vivo rates of protein synthesis and ion transport increased ∼50% under acidification. Importantly, the in vivo physiological increases in ion transport were not predicted from total enzyme activity or gene expression. Under acidification, the increased rates of protein synthesis and ion transport that were sustained in growing larvae collectively accounted for the majority of available ATP (84%). In contrast, embryos and prefeeding and unfed larvae in control treatments allocated on average only 40% of ATP to these same two processes. Understanding the biochemical strategies for accommodating increases in metabolic energy demand and their biological limitations can serve as a quantitative basis for assessing sublethal effects of global change. Variation in the ability to allocate ATP differentially among essential functions may be a key basis of resilience to ocean acidification and other compounding environmental stressors. PMID:25825763

  12. Characterization of the Deleted in Autism 1 Protein Family: Implications for Studying Cognitive Disorders

    PubMed Central

    Aziz, Azhari; Harrop, Sean P.; Bishop, Naomi E.

    2011-01-01

    Autism spectrum disorders (ASDs) are a group of commonly occurring, highly-heritable developmental disabilities. Human genes c3orf58 or Deleted In Autism-1 (DIA1) and cXorf36 or Deleted in Autism-1 Related (DIA1R) are implicated in ASD and mental retardation. Both gene products encode signal peptides for targeting to the secretory pathway. As evolutionary medicine has emerged as a key tool for understanding increasing numbers of human diseases, we have used an evolutionary approach to study DIA1 and DIA1R. We found DIA1 conserved from cnidarians to humans, indicating DIA1 evolution coincided with the development of the first primitive synapses. Nematodes lack a DIA1 homologue, indicating Caenorhabditis elegans is not suitable for studying all aspects of ASD etiology, while zebrafish encode two DIA1 paralogues. By contrast to DIA1, DIA1R was found exclusively in vertebrates, with an origin coinciding with the whole-genome duplication events occurring early in the vertebrate lineage, and the evolution of the more complex vertebrate nervous system. Strikingly, DIA1R was present in schooling fish but absent in fish that have adopted a more solitary lifestyle. An additional DIA1-related gene we named DIA1-Like (DIA1L), lacks a signal peptide and is restricted to the genomes of the echinoderm Strongylocentrotus purpuratus and cephalochordate Branchiostoma floridae. Evidence for remarkable DIA1L gene expansion was found in B. floridae. Amino acid alignments of DIA1 family gene products revealed a potential Golgi-retention motif and a number of conserved motifs with unknown function. Furthermore, a glycine and three cysteine residues were absolutely conserved in all DIA1-family proteins, indicating a critical role in protein structure and/or function. We have therefore identified a new metazoan protein family, the DIA1-family, and understanding the biological roles of DIA1-family members will have implications for our understanding of autism and mental retardation. PMID

  13. Distribution and evolution of the serine/aspartate racemase family in invertebrates.

    PubMed

    Uda, Kouji; Abe, Keita; Dehara, Yoko; Mizobata, Kiriko; Sogawa, Natsumi; Akagi, Yuki; Saigan, Mai; Radkov, Atanas D; Moe, Luke A

    2016-02-01

    Free D-amino acids have been found in various invertebrate phyla, while amino acid racemase genes have been identified in few species. The purpose of this study is to elucidate the distribution, function, and evolution of amino acid racemases in invertebrate animals. We searched the GenBank databases, and found 11 homologous serine racemase genes from eight species in eight different invertebrate phyla. The cloned genes were identified based on their maximum activity as Acropora millepora (Cnidaria) serine racemase (SerR) and aspartate racemase (AspR), Caenorhabditis elegans (Nematoda) SerR, Capitella teleta (Annelida) SerR, Crassostrea gigas (Mollusca) SerR and AspR, Dugesia japonica (Platyhelminthes) SerR, Milnesium tardigradum (Tardigrada) SerR, Penaeus monodon (Arthropoda) SerR and AspR and Strongylocentrotus purpuratus (Echinodermata) AspR. We found that Acropora, Aplysia, Capitella, Crassostrea and Penaeus had two amino acid racemase paralogous genes and these paralogous genes have evolved independently by gene duplication at their recent ancestral species. The transcriptome analyses using available SRA data and enzyme kinetic data suggested that these paralogous genes are expressed in different tissues and have different functions in vivo. Phylogenetic analyses clearly indicated that animal SerR and AspR are not separated by their particular racemase functions and form a serine/aspartate racemase family cluster. Our results revealed that SerR and AspR are more widely distributed among invertebrates than previously known. Moreover, we propose that the triple serine loop motif at amino acid positions 150-152 may be responsible for the large aspartate racemase activity and the AspR evolution from SerR. PMID:26352274

  14. Stormwater toxicity in Chollas Creek and San Diego Bay, California.

    PubMed

    Schiff, Kenneth; Bay, Steven; Diehl, Dario

    2003-01-01

    Stormwater discharges from Chollas Creek, a tributary of San Diego Bay, have been shown to be toxic to aquatic life. The primary objective of this study was to provide the linkage between in-channel measurements and potential impairments in the receiving waters of San Diego Bay. This study addressed this objective within the context of four questions: (1) How much area in San Diego Bay is affected by the discharge plume from Chollas Creek during wet-weather conditions?; (2) How much of the wet-weather discharge plume is toxic to marine aquatic life?; (3) How toxic is this area within the wet-weather discharge plume?; and (4) What are the constituent(s) responsible for the observed toxicity in the wet-weather plume? The stormwater plume emanating from Chollas Creek was dynamic, covering areas up to 2.25 km2. Approximately half of the plume was estimated to be toxic to marine life, based upon the results of purple sea urchin (Strongylocentroutus purpuratus) fertilization tests. The area nearest the creek mouth was the most toxic (NOEC = 3 to 12% plume sample), and the toxicity decreased with distance from the creek mouth. The toxicity of plume samples was directly proportional to the magnitude of plume mixing and dilution until, once outside the plume margin, no toxicity was observed. Trace metals, most likely zinc, were responsible for the observed plume toxicity based upon toxicity identification evaluations (TIEs). Zinc was also the constituent identified from in-channel samples of Chollas Creek stormwater using TIEs on the storms sampled in this study, and in storms sampled during the previous storm season. PMID:12620010

  15. Aggregation of sea urchin phagocytes is augmented in vitro by lipopolysaccharide.

    PubMed

    Majeske, Audrey J; Bayne, Christopher J; Smith, L Courtney

    2013-01-01

    Development of protocols and media for culturing immune cells from marine invertebrates has not kept pace with advancements in mammalian immune cell culture, the latter having been driven by the need to understand the causes of and develop therapies for human and animal diseases. However, expansion of the aquaculture industry and the diseases that threaten these systems creates the need to develop cell and tissue culture methods for marine invertebrates. Such methods will enable us to better understand the causes of disease outbreaks and to develop means to avoid and remedy epidemics. We report a method for the short-term culture of phagocytes from the purple sea urchin, Strongylocentrotus purpuratus, by modifying an approach previously used to culture cells from another sea urchin species. The viability of cultured phagocytes from the purple sea urchin decreases from 91.6% to 57% over six days and phagocyte morphology changes from single cells to aggregates leading to the formation of syncytia-like structures. This process is accelerated in the presence of lipopolysaccharide suggesting that phagocytes are capable of detecting this molecular pattern in culture conditions. Sea urchin immune response proteins, called Sp185/333, are expressed on the surface of a subset of phagocytes and have been associated with syncytia-like structures. We evaluated their expression in cultured phagocytes to determine their possible role in cell aggregation and in the formation of syncytia-like structures. Between 0 and 3 hr, syncytia-like structures were observed in cultures when only ~10% of the cells were positive for Sp185/333 proteins. At 24 hr, ~90% of the nuclei were Sp185/333-positive when all of the phagocytes had aggregated into syncytia-like structures. Consequently, we conclude that the Sp185/333 proteins do not have a major role in initiating the aggregation of cultured phagocytes, however the Sp185/333 proteins are associated with the clustered nuclei within the

  16. Comparative Study of Regulatory Circuits in Two Sea Urchin Species Reveals Tight Control of Timing and High Conservation of Expression Dynamics

    PubMed Central

    Gildor, Tsvia; Ben-Tabou de-Leon, Smadar

    2015-01-01

    Accurate temporal control of gene expression is essential for normal development and must be robust to natural genetic and environmental variation. Studying gene expression variation within and between related species can delineate the level of expression variability that development can tolerate. Here we exploit the comprehensive model of sea urchin gene regulatory networks and generate high-density expression profiles of key regulatory genes of the Mediterranean sea urchin, Paracentrotus lividus (Pl). The high resolution of our studies reveals highly reproducible gene initiation times that have lower variation than those of maximal mRNA levels between different individuals of the same species. This observation supports a threshold behavior of gene activation that is less sensitive to input concentrations. We then compare Mediterranean sea urchin gene expression profiles to those of its Pacific Ocean relative, Strongylocentrotus purpuratus (Sp). These species shared a common ancestor about 40 million years ago and show highly similar embryonic morphologies. Our comparative analyses of five regulatory circuits operating in different embryonic territories reveal a high conservation of the temporal order of gene activation but also some cases of divergence. A linear ratio of 1.3-fold between gene initiation times in Pl and Sp is partially explained by scaling of the developmental rates with temperature. Scaling the developmental rates according to the estimated Sp-Pl ratio and normalizing the expression levels reveals a striking conservation of relative dynamics of gene expression between the species. Overall, our findings demonstrate the ability of biological developmental systems to tightly control the timing of gene activation and relative dynamics and overcome expression noise induced by genetic variation and growth conditions. PMID:26230518

  17. The chronic toxicity of molybdate to marine organisms. I. Generating reliable effects data.

    PubMed

    Heijerick, D G; Regoli, L; Stubblefield, W

    2012-07-15

    A scientific research program was initiated by the International Molybdenum Association (IMOA) which addressed identified gaps in the environmental toxicity data for the molybdate ion (MoO(4)(2-)). These gaps were previously identified during the preparation of EU-REACH-dossiers for different molybdenum compounds (European Union regulation on Registration, Evaluation, Authorization and Restriction of Chemical substances; EC, 2006). Evaluation of the open literature identified few reliable marine ecotoxicological data that could be used for deriving a Predicted No-Effect Concentration (PNEC) for the marine environment. Rather than calculating a PNEC(marine) using the assessment factor methodology on a combined freshwater/marine dataset, IMOA decided to generate sufficient reliable marine chronic data to permit derivation of a PNEC by means of the more scientifically robust species sensitivity distribution (SSD) approach (also called the statistical extrapolation approach). Nine test species were chronically exposed to molybdate (added as sodium molybdate dihydrate, Na(2)MoO(4)·2H(2)O) according to published standard testing guidelines that are acceptable for a broad range of regulatory purposes. The selected test organisms were representative for typical marine trophic levels: micro-algae/diatom (Phaeodactylum tricornutum, Dunaliella tertiolecta), macro-alga (Ceramium tenuicorne), mysids (Americamysis bahia), copepod (Acartia tonsa), fish (Cyprinodon variegatus), echinoderms (Dendraster exentricus, Strongylocentrotus purpuratus) and molluscs (Mytilus edulis, Crassostrea gigas). Available NOEC/EC(10) levels ranged between 4.4 mg Mo/L (blue mussel M. edulis) and 1174 mg Mo/L (oyster C. gigas). Using all available reliable marine chronic effects data that are currently available, a HC(5,50%) (median hazardous concentration affecting 5% of the species) of 5.74(mg Mo)/L was derived with the statistical extrapolation approach, a value that can be used for national and

  18. Acute effects of non-weathered and weathered crude oil and dispersant associated with the Deepwater Horizon incident on the development of marine bivalve and echinoderm larvae.

    PubMed

    Stefansson, Emily S; Langdon, Chris J; Pargee, Suzanne M; Blunt, Susanna M; Gage, Susan J; Stubblefield, William A

    2016-08-01

    Acute toxicity tests (48-96-h duration) were conducted with larvae of 2 echinoderm species (Strongylocentrotus purpuratus and Dendraster excentricus) and 4 bivalve mollusk species (Crassostrea virginica, Crassostrea gigas, Mytilus galloprovincialis, and Mercenaria mercenaria). Developing larvae were exposed to water-accommodated fractions (WAFs) and chemically enhanced water-accommodated fractions (CEWAFs) of fresh and weathered oils collected from the Gulf of Mexico during the Deepwater Horizon incident. The WAFs (oils alone), CEWAFs (oils plus Corexit 9500A dispersant), and WAFs of Corexit alone were prepared using low-energy mixing. The WAFs of weathered oils had no effect on survival and development of echinoderm and bivalve larvae, whereas WAFs of fresh oils showed adverse effects on larval development. Similar toxicities were observed for weathered oil CEWAFs and WAFs prepared with Corexit alone for oyster (C. gigas and C. virginica) larvae, which were the most sensitive of the tested invertebrate species to Corexit. Mean 10% effective concentration values for total polycyclic aromatic hydrocarbons and dipropylene glycol n-butyl ether (a marker for Corexit) in the present study were higher than all concentrations reported in nearshore field samples collected during and after the Deepwater Horizon incident. The results suggest that water-soluble fractions of weathered oils and Corexit dispersant associated with the Deepwater Horizon incident had limited, if any, acute impacts on nearshore larvae of eastern oysters and clams, as well as other organisms with similar sensitivities to those of test species in the present study; however, exposure to sediments and long-term effects were not evaluated. Environ Toxicol Chem 2016;35:2016-2028. © 2016 SETAC. PMID:26749266

  19. Phylogeny of Echinoderm Hemoglobins

    PubMed Central

    Christensen, Ana B.; Herman, Joseph L.; Elphick, Maurice R.; Kober, Kord M.; Janies, Daniel; Linchangco, Gregorio; Semmens, Dean C.; Bailly, Xavier; Vinogradov, Serge N.; Hoogewijs, David

    2015-01-01

    Background Recent genomic information has revealed that neuroglobin and cytoglobin are the two principal lineages of vertebrate hemoglobins, with the latter encompassing the familiar myoglobin and α-globin/β-globin tetramer hemoglobin, and several minor groups. In contrast, very little is known about hemoglobins in echinoderms, a phylum of exclusively marine organisms closely related to vertebrates, beyond the presence of coelomic hemoglobins in sea cucumbers and brittle stars. We identified about 50 hemoglobins in sea urchin, starfish and sea cucumber genomes and transcriptomes, and used Bayesian inference to carry out a molecular phylogenetic analysis of their relationship to vertebrate sequences, specifically, to assess the hypothesis that the neuroglobin and cytoglobin lineages are also present in echinoderms. Results The genome of the sea urchin Strongylocentrotus purpuratus encodes several hemoglobins, including a unique chimeric 14-domain globin, 2 androglobin isoforms and a unique single androglobin domain protein. Other strongylocentrotid genomes appear to have similar repertoires of globin genes. We carried out molecular phylogenetic analyses of 52 hemoglobins identified in sea urchin, brittle star and sea cucumber genomes and transcriptomes, using different multiple sequence alignment methods coupled with Bayesian and maximum likelihood approaches. The results demonstrate that there are two major globin lineages in echinoderms, which are related to the vertebrate neuroglobin and cytoglobin lineages. Furthermore, the brittle star and sea cucumber coelomic hemoglobins appear to have evolved independently from the cytoglobin lineage, similar to the evolution of erythroid oxygen binding globins in cyclostomes and vertebrates. Conclusion The presence of echinoderm globins related to the vertebrate neuroglobin and cytoglobin lineages suggests that the split between neuroglobins and cytoglobins occurred in the deuterostome ancestor shared by echinoderms and

  20. Majority of divergence between closely related DNA samples is due to indels

    PubMed Central

    Britten, Roy J.; Rowen, Lee; Williams, John; Cameron, R. Andrew

    2003-01-01

    It was recently shown that indels are responsible for more than twice as many unmatched nucleotides as are base substitutions between samples of chimpanzee and human DNA. A larger sample has now been examined and the result is similar. The number of indels is ≈1/12th of the number of base substitutions and the average length of the indels is 36 nt, including indels up to 10 kb. The ratio (Ru) of unpaired nucleotides attributable to indels to those attributable to substitutions is 3.0 for this 2 million-nt chimp DNA sample compared with human. There is similar evidence of a large value of Ru for sea urchins from the polymorphism of a sample of Strongylocentrotus purpuratus DNA (Ru = 3–4). Other work indicates that similarly, per nucleotide affected, large differences are seen for indels in the DNA polymorphism of the plant Arabidopsis thaliana (Ru = 51). For the insect Drosophila melanogaster a high value of Ru (4.5) has been determined. For the nematode Caenorhabditis elegans the polymorphism data are incomplete but high values of Ru are likely. Comparison of two strains of Escherichia coli O157:H7 shows a preponderance of indels. Because these six examples are from very distant systematic groups the implication is that in general, for alignments of closely related DNA, indels are responsible for many more unmatched nucleotides than are base substitutions. Human genetic evidence suggests that indels are a major source of gene defects, indicating that indels are a significant source of evolutionary change. PMID:12672966

  1. Specific functions of the Wnt signaling system in gene regulatory networks throughout the early sea urchin embryo

    PubMed Central

    Cui, Miao; Siriwon, Natnaree; Li, Enhu; Davidson, Eric H.; Peter, Isabelle S.

    2014-01-01

    Wnt signaling affects cell-fate specification processes throughout embryonic development. Here we take advantage of the well-studied gene regulatory networks (GRNs) that control pregastrular sea urchin embryogenesis to reveal the gene regulatory functions of the entire Wnt-signaling system. Five wnt genes, three frizzled genes, two secreted frizzled-related protein 1 genes, and two Dickkopf genes are expressed in dynamic spatial patterns in the pregastrular embryo of Strongylocentrotus purpuratus. We present a comprehensive analysis of these genes in each embryonic domain. Total functions of the Wnt-signaling system in regulatory gene expression throughout the embryo were studied by use of the Porcupine inhibitor C59, which interferes with zygotic Wnt ligand secretion. Morpholino-mediated knockdown of each expressed Wnt ligand demonstrated that individual Wnt ligands are functionally distinct, despite their partially overlapping spatial expression. They target specific embryonic domains and affect particular regulatory genes. The sum of the effects of blocking expression of individual wnt genes is shown to equal C59 effects. Remarkably, zygotic Wnt-signaling inputs are required for only three general aspects of embryonic specification: the broad activation of endodermal GRNs, the regional specification of the immediately adjacent stripe of ectoderm, and the restriction of the apical neurogenic domain. All Wnt signaling in this pregastrular embryo is short range (and/or autocrine). Furthermore, we show that the transcriptional drivers of wnt genes execute important specification functions in the embryonic domains targeted by the ligands, thus connecting the expression and function of wnt genes by encoded cross-regulatory interactions within the specific regional GRNs. PMID:25385617

  2. Species sensitivity distribution evaluation for chronic nickel toxicity to marine organisms.

    PubMed

    DeForest, David K; Schlekat, Christian E

    2013-10-01

    In Europe, the European Union's Existing Substances Regulation (EEC 793/93), the REACH Regulation, and Water Framework Directive all share common guidance for conducting environmental effects assessments, which can be further used to derive predicted no effect concentrations (PNECs) and environmental quality standards (EQS) for chemical substances. To meet the criteria for using a species sensitivity distribution (SSD) in the effects assessment of Ni for marine organisms, chronic toxicity data from the published scientific literature were augmented with toxicity testing of several additional marine species including: a unicellular alga (Dunalliela tertiolecta), a diatom (Skeletonema costatum), 2 macroalgae (Champia parvula, Macrocystis pyrifera), 2 mollusks (Crassostrea gigas, Mytilus galloprovincialis), 2 echinoderms (Dendraster excentricus, Strongylocentrotus purpuratus), a polychaete (Neanthes arenaceodentata), and a fish (Cyprinodon variegatus). Based on this updated database, which includes chronic Ni toxicity data for a total of 17 marine species, HC5 values (hazardous concentrations to 5% of the species) were derived using an SSD. The most sensitive species is a tropical sea urchin from the Caribbean region, Diadema antillarum, which has an EC10 that is approximately 6-fold less than the EC10 for the second most sensitive species tested. There is some uncertainty in the representativeness of D. antillarum to temperate European marine waters because 1) a European sea urchin species (Paracentrotus lividus) is approximately 48-fold less sensitive to Ni, and (2) ambient marine Ni concentrations in at least some European waters closely approach the D. antillarum EC10. The HC5 values with and without D. antillarum included in the SSD are 3.9 and 20.9 μg/L, respectively. Site-specific toxicity testing with local species may be warranted for locations where Ni concentrations fall between the range in HC5s of 3.9 to 20.9 μg/L. PMID:23553986

  3. Single sea urchin phagocytes express messages of a single sequence from the diverse Sp185/333 gene family in response to bacterial challenge.

    PubMed

    Majeske, Audrey J; Oren, Matan; Sacchi, Sandro; Smith, L Courtney

    2014-12-01

    Immune systems in animals rely on fast and efficient responses to a wide variety of pathogens. The Sp185/333 gene family in the purple sea urchin, Strongylocentrotus purpuratus, consists of an estimated 50 (±10) members per genome that share a basic gene structure but show high sequence diversity, primarily due to the mosaic appearance of short blocks of sequence called elements. The genes show significantly elevated expression in three subpopulations of phagocytes responding to marine bacteria. The encoded Sp185/333 proteins are highly diverse and have central effector functions in the immune system. In this study we report the Sp185/333 gene expression in single sea urchin phagocytes. Sea urchins challenged with heat-killed marine bacteria resulted in a typical increase in coelomocyte concentration within 24 h, which included an increased proportion of phagocytes expressing Sp185/333 proteins. Phagocyte fractions enriched from coelomocytes were used in limiting dilutions to obtain samples of single cells that were evaluated for Sp185/333 gene expression by nested RT-PCR. Amplicon sequences showed identical or nearly identical Sp185/333 amplicon sequences in single phagocytes with matches to six known Sp185/333 element patterns, including both common and rare element patterns. This suggested that single phagocytes show restricted expression from the Sp185/333 gene family and infers a diverse, flexible, and efficient response to pathogens. This type of expression pattern from a family of immune response genes in single cells has not been identified previously in other invertebrates. PMID:25355922

  4. Beyond BLASTing: tertiary and quaternary structure analysis helps identify major vault proteins.

    PubMed

    Daly, Toni K; Sutherland-Smith, Andrew J; Penny, David

    2013-01-01

    We examine the advantages of going beyond sequence similarity and use both protein three-dimensional (3D) structure prediction and then quaternary structure (docking) of inferred 3D structures to help evaluate whether comparable sequences can fold into homologous structures with sufficient lateral associations for quaternary structure formation. Our test case is the major vault protein (MVP) that oligomerizes in multiple copies to form barrel-like vault particles and is relatively widespread among eukaryotes. We used the iterative threading assembly refinement server (I-TASSER) to predict whether putative MVP sequences identified by BLASTp and PSI Basic Local Alignment Search Tool are structurally similar to the experimentally determined rodent MVP tertiary structures. Then two identical predicted quaternary structures from I-TASSER are analyzed by RosettaDock to test whether a pair-wise association occurs, and hence whether the oligomeric vault complex is likely to form for a given MVP sequence. Positive controls for the method are the experimentally determined rat (Rattus norvegicus) vault X-ray crystal structure and the purple sea urchin (Strongylocentrotus purpuratus) MVP sequence that forms experimentally observed vaults. These and two kinetoplast MVP structural homologs were predicted with high confidence value, and RosettaDock predicted that these MVP sequences would dock laterally and therefore could form oligomeric vaults. As the negative control, I-TASSER did not predict an MVP-like structure from a randomized rat MVP sequence, even when constrained to the rat MVP crystal structure (PDB:2ZUO), thus further validating the method. The protocol identified six putative homologous MVP sequences in the heterobolosean Naegleria gruberi within the excavate kingdom. Two of these sequences are predicted to be structurally similar to rat MVP, despite being in excess of 300 residues shorter. The method can be used generally to help test predictions of homology via

  5. Calbindin-D32k Is Localized to a Subpopulation of Neurons in the Nervous System of the Sea Cucumber Holothuria glaberrima (Echinodermata)

    PubMed Central

    Díaz-Balzac, Carlos A.; Lázaro-Peña, María I.; García-Rivera, Enrique M.; González, Carlos I.; García-Arrarás, José E.

    2012-01-01

    Members of the calbindin subfamily serve as markers of subpopulations of neurons within the vertebrate nervous system. Although markers of these proteins are widely available and used, their application to invertebrate nervous systems has been very limited. In this study we investigated the presence and distribution of members of the calbindin subfamily in the sea cucumber Holothuria glaberrima (Selenka, 1867). Immunohistological experiments with antibodies made against rat calbindin 1, parvalbumin, and calbindin 2, showed that these antibodies labeled cells and fibers within the nervous system of H. glaberrima. Most of the cells and fibers were co-labeled with the neural-specific marker RN1, showing their neural specificity. These were distributed throughout all of the nervous structures, including the connective tissue plexi of the body wall and podia. Bioinformatics analyses of the possible antigen recognized by these markers showed that a calbindin 2-like protein present in the sea urchin Strongylocentrotus purpuratus, corresponded to the calbindin-D32k previously identified in other invertebrates. Western blots with anti-calbindin 1 and anti-parvalbumin showed that these markers recognized an antigen of approximately 32 kDa in homogenates of radial nerve cords of H. glaberrima and Lytechinus variegatus. Furthermore, immunoreactivity with anti-calbindin 1 and anti-parvalbumin was obtained to a fragment of calbindin-D32k of H. glaberrima. Our findings suggest that calbindin-D32k is present in invertebrates and its sequence is more similar to the vertebrate calbindin 2 than to calbindin 1. Thus, characterization of calbindin-D32k in echinoderms provides an important view of the evolution of this protein family and represents a valuable marker to study the nervous system of invertebrates. PMID:22412907

  6. Search for eukaryotic motility proteins in spirochetes: immunological detection of a tektin-like protein in Spirochaeta halophila.

    PubMed

    Barth, A L; Stricker, J A; Margulis, L

    1991-01-01

    The serial endosymbiotic theory (SET) for the spirochete origin of undulipodia (cilia and eukaryotic flagella) predicts that a greater number of axonemal proteins will show homology to spirochete than to other bacterial proteins. To continue testing, the SET proteins associated with eukaryotic motility (tektin, centrin and calmodulin) were sought in Spirochaeta halophila. Western blot immunological detection techniques (for tektin and centrin) and two-dimensional gel analysis (for calmodulin) were used. Tektins are filamentous proteins extending the length of the axoneme in sperm tails and other undulipodia. Whole cell extracts of S. halophila were probed with antibodies made against three sea urchin (Lytechinus pictus) sperm axonemal tektins (tektins A, B, and C). In the spirochetes, one tektin-like protein was detected as a band on Western blots (a C tektin.) An antibody made against Lytechinus pictus sperm tail axonemes, affinity-purified against the C tektin of another sea urchin, Stronglyocentrotus purpuratus, bound to a 30 kDa protein from Spirochaeta halophila. The C tektin epitope was not detected in Escherichia coli. Both the poly- and monoclonal anti-centrin antibodies cross-reacted with multiple proteins in the control alga (Tetraselmis striata) and in the putatively negative control bacterium E. coli. No cross reaction was seen between any anti-centrin antibody and S. halophila. Neither did a two-dimensional gel analysis reveal the presence of calmodulin in these spirochetes or in the two other prokaryotes tested (Spiroplasma citri, Acholeplasma laidlawii). Although neither centrin nor calmodulin were detected, a 30 kDa tektin-like protein apparently is present in these spirochetes. PMID:1863720

  7. Curve fitting toxicity test data: Which comes first, the dose response or the model?

    SciTech Connect

    Gully, J.; Baird, R.; Bottomley, J.

    1995-12-31

    The probit model frequently does not fit the concentration-response curve of NPDES toxicity test data and non-parametric models must be used instead. The non-parametric models, trimmed Spearman-Karber, IC{sub p}, and linear interpolation, all require a monotonic concentration-response. Any deviation from a monotonic response is smoothed to obtain the desired concentration-response characteristics. Inaccurate point estimates may result from such procedures and can contribute to imprecision in replicate tests. The following study analyzed reference toxicant and effluent data from giant kelp (Macrocystis pyrifera), purple sea urchin (Strongylocentrotus purpuratus), red abalone (Haliotis rufescens), and fathead minnow (Pimephales promelas) bioassays using commercially available curve fitting software. The purpose was to search for alternative parametric models which would reduce the use of non-parametric models for point estimate analysis of toxicity data. Two non-linear models, power and logistic dose-response, were selected as possible alternatives to the probit model based upon their toxicological plausibility and ability to model most data sets examined. Unlike non-parametric procedures, these and all parametric models can be statistically evaluated for fit and significance. The use of the power or logistic dose response models increased the percentage of parametric model fits for each protocol and toxicant combination examined. The precision of the selected non-linear models was also compared with the EPA recommended point estimation models at several effect.levels. In general, precision of the alternative models was equal to or better than the traditional methods. Finally, use of the alternative models usually produced more plausible point estimates in data sets where the effects of smoothing and non-parametric modeling made the point estimate results suspect.

  8. Base excision DNA repair in the embryonic development of the sea urchin, Strongylocentrotus intermedius.

    PubMed

    Torgasheva, Natalya A; Menzorova, Natalya I; Sibirtsev, Yurii T; Rasskazov, Valery A; Zharkov, Dmitry O; Nevinsky, Georgy A

    2016-06-21

    In actively proliferating cells, such as the cells of the developing embryo, DNA repair is crucial for preventing the accumulation of mutations and synchronizing cell division. Sea urchin embryo growth was analyzed and extracts were prepared. The relative activity of DNA polymerase, apurinic/apyrimidinic (AP) endonuclease, uracil-DNA glycosylase, 8-oxoguanine-DNA glycosylase, and other glycosylases was analyzed using specific oligonucleotide substrates of these enzymes; the reaction products were resolved by denaturing 20% polyacrylamide gel electrophoresis. We have characterized the profile of several key base excision repair activities in the developing embryos (2 blastomers to mid-pluteus) of the grey sea urchin, Strongylocentrotus intermedius. The uracil-DNA glycosylase specific activity sharply increased after blastula hatching, whereas the specific activity of 8-oxoguanine-DNA glycosylase steadily decreased over the course of the development. The AP-endonuclease activity gradually increased but dropped at the last sampled stage (mid-pluteus 2). The DNA polymerase activity was high at the first cleavage division and then quickly decreased, showing a transient peak at blastula hatching. It seems that the developing sea urchin embryo encounters different DNA-damaging factors early in development within the protective envelope and later as a free-floating larva, with hatching necessitating adaptation to the shift in genotoxic stress conditions. No correlation was observed between the dynamics of the enzyme activities and published gene expression data from developing congeneric species, S. purpuratus. The results suggest that base excision repair enzymes may be regulated in the sea urchin embryos at the level of covalent modification or protein stability. PMID:27158700

  9. Mature maternal mRNAs are longer than zygotic ones and have complex degradation kinetics in sea urchin.

    PubMed

    Gildor, Tsvia; Malik, Assaf; Sher, Noa; Ben-Tabou de-Leon, Smadar

    2016-06-01

    Early in embryogenesis, maternally deposited transcripts are degraded and new zygotic transcripts are generated during the maternal to zygotic transition. Recent works have shown that early zygotic transcripts are short compared to maternal transcripts, in zebrafish and Drosophila species. The reduced zygotic transcript length was attributed to the short cell cycle in these organisms that prevents the transcription of long primary transcripts (intron delay). Here we study the length of maternal mRNAs and their degradation kinetics in two sea urchin species to further the understanding of maternal gene usage and processing. Early zygotic primary transcripts and mRNAs are shorter than maternal ones in the sea urchin, Strongylocentrotus purpuratus. Yet, while primary transcripts length increases when cell cycle lengthens, typical for intron delay, the relatively short length of zygotic mRNAs is consistent. The enhanced mRNA length is due to significantly longer maternal open reading frames and 3'UTRs compared to the zygotic lengths, a ratio that does not change with developmental time. This implies unique usage of both coding sequences and regulatory information in the maternal stage compared to the zygotic stages. We extracted the half-lifetimes due to maternal and zygotic degradation mechanisms from high-density time course of a set of maternal mRNAs in Paracentrotus lividus. The degradation rates due to maternal and zygotic degradation mechanisms are not correlated, indicating that these mechanisms are independent and relay on different regulatory information. Our studies illuminate specific structural and kinetic properties of sea urchin maternal mRNAs that might be broadly shared by other organisms. PMID:27085752

  10. Two cis elements collaborate to spatially repress transcription from a sea urchin promoter

    NASA Technical Reports Server (NTRS)

    Frudakis, T. N.; Wilt, F.

    1995-01-01

    The expression pattern of many territory-specific genes in metazoan embryos is maintained by an active process of negative spatial regulation. However, the mechanism of this strategy of gene regulation is not well understood in any system. Here we show that reporter constructs containing regulatory sequence for the SM30-alpha gene of Stronglyocentrotus purpuratus are expressed in a pattern congruent with that of the endogenous SM30 gene(s), largely as a result of active transcriptional repression in cell lineages in which the gene is not normally expressed. Chloramphenicol acetyl transferase assays of deletion constructs from the 2600-bp upstream region showed that repressive elements were present in the region from -1628 to -300. In situ hybridization analysis showed that the spatial fidelity of expression was severely compromised when the region from -1628 to -300 was deleted. Two highly repetitive sequence motifs, (G/A/C)CCCCT and (T/C)(T/A/C)CTTTT(T/A/C), are present in the -1628 to -300 region. Representatives of these elements were analyzed by gel mobility shift experiments and were found to interact specifically with protein in crude nuclear extracts. When oligonucleotides containing either sequence element were co-injected with a correctly regulated reporter as potential competitors, the reporter was expressed in inappropriate cells. When composite oligonucleotides, containing both sequence elements, were fused to a misregulated reporter, the expression of the reporter in inappropriate cells was suppressed. Comparison of composite oligonucleotides with oligonucleotides containing single constituent elements show that both sequence elements are required for effective spatial regulation. Thus, both individual elements are required, but only a composite element containing both elements is sufficient to function as a tissue-specific repressive element.

  11. Regulation of membrane fusion and secretory events in the sea urchin embryo

    SciTech Connect

    Roe, J.L.

    1990-01-01

    Membrane fusion and secretory events play a key role in fertilization and early development in the sea urchin embryo. To investigate the mechanism of membrane fusion, the effect of inhibitors of metalloendoprotease activity was studied on two model systems of cell fusion; fertilization and spiculogenesis by primary mesenchyme cells in the embryo. Both the zinc chelator, 1,10-phenanthroline, and peptide metalloprotease substrates were found to inhibit both fertilization and gamete fusion, while peptides that are not substrates of metalloproteases did not affect either process. Primary mesenchyme cells form the larval skeleton in the embryo by deposition of mineral and an organic matrix into a syncytial cavity formed by fusion of filopodia of these cells. Metalloprotease inhibitors were found to inhibit spiculogenesis both in vivo and in cultures of isolated primary mesenchyme cells, and the activity of a metalloprotease of the appropriate specificity was found in the primary mesenchyme cells. These two studies implicate the activity of a metalloprotease in a necessary step in membrane fusion. Following fertilization, exocytosis of the cortical granules results in the formation of the fertilization envelope and the hyaline layer, that surround the developing embryo. The hatching enzyme is secreted by the blastula stage sea urchin embryo, which proteolyzes the fertilization envelope surrounding the embryo, allowing the embryo to hatch. Using an assay that measures {sup 125}I-fertilization envelope degradation, the hatching enzyme was identified as a 33 kDa metalloprotease, and was purified by ion-exchange and affinity chromatography from the hatching media of Strongylocentrotus purpuratus embryos. The hatching enzyme showed a substrate preference for only a minor subset of fertilization envelope proteins.

  12. An Elk transcription factor is required for Runx-dependent survival signaling in the sea urchin embryo.

    PubMed

    Rizzo, Francesca; Coffman, James A; Arnone, Maria Ina

    2016-08-01

    Elk proteins are Ets family transcription factors that regulate cell proliferation, survival, and differentiation in response to ERK (extracellular-signal regulated kinase)-mediated phosphorylation. Here we report the embryonic expression and function of Sp-Elk, the single Elk gene of the sea urchin Strongylocentrotus purpuratus. Sp-Elk is zygotically expressed throughout the embryo beginning at late cleavage stage, with peak expression occurring at blastula stage. Morpholino antisense-mediated knockdown of Sp-Elk causes blastula-stage developmental arrest and embryo disintegration due to apoptosis, a phenotype that is rescued by wild-type Elk mRNA. Development is also rescued by Elk mRNA encoding a serine to aspartic acid substitution (S402D) that mimics ERK-mediated phosphorylation of a conserved site that enhances DNA binding, but not by Elk mRNA encoding an alanine substitution at the same site (S402A). This demonstrates both that the apoptotic phenotype of the morphants is specifically caused by Elk depletion, and that phosphorylation of serine 402 of Sp-Elk is critical for its anti-apoptotic function. Knockdown of Sp-Elk results in under-expression of several regulatory genes involved in cell fate specification, cell cycle control, and survival signaling, including the transcriptional regulator Sp-Runt-1 and its target Sp-PKC1, both of which were shown previously to be required for cell survival during embryogenesis. Both Sp-Runt-1 and Sp-PKC1 have sequences upstream of their transcription start sites that specifically bind Sp-Elk. These results indicate that Sp-Elk is the signal-dependent activator of a feed-forward gene regulatory circuit, consisting also of Sp-Runt-1 and Sp-PKC1, which actively suppresses apoptosis in the early embryo. PMID:27235147

  13. Genetics of gene expression responses to temperature stress in a sea urchin gene network.

    PubMed

    Runcie, Daniel E; Garfield, David A; Babbitt, Courtney C; Wygoda, Jennifer A; Mukherjee, Sayan; Wray, Gregory A

    2012-09-01

    Stress responses play an important role in shaping species distributions and robustness to climate change. We investigated how stress responses alter the contribution of additive genetic variation to gene expression during development of the purple sea urchin, Strongylocentrotus purpuratus, under increased temperatures that model realistic climate change scenarios. We first measured gene expression responses in the embryos by RNA-seq to characterize molecular signatures of mild, chronic temperature stress in an unbiased manner. We found that an increase from 12 to 18 °C caused widespread alterations in gene expression including in genes involved in protein folding, RNA processing and development. To understand the quantitative genetic architecture of this response, we then focused on a well-characterized gene network involved in endomesoderm and ectoderm specification. Using a breeding design with wild-caught individuals, we measured genetic and gene-environment interaction effects on 72 genes within this network. We found genetic or maternal effects in 33 of these genes and that the genetic effects were correlated in the network. Fourteen network genes also responded to higher temperatures, but we found no significant genotype-environment interactions in any of the genes. This absence may be owing to an effective buffering of the temperature perturbations within the network. In support of this hypothesis, perturbations to regulatory genes did not affect the expression of the genes that they regulate. Together, these results provide novel insights into the relationship between environmental change and developmental evolution and suggest that climate change may not expose large amounts of cryptic genetic variation to selection in this species. PMID:22856327

  14. A G/C-rich DNA-regulatory element controls positive expression of the sea urchin Lytechinus pictus aboral ectoderm-specific LpS1 gene.

    PubMed

    Wang, W; Klein, W H

    1996-02-01

    The LpS1 beta gene of Lytechinus pictus is activated at the late cleavage stage about 12 hr after fertilization. LpS1 beta transcripts accumulate exclusively in aboral ectoderm lineages. LpS1 beta is thus a classic example of a gene whose expression is tightly controlled both temporally and spatially during early development. Previous studies on the LpS1 beta promoter identified two G-string DNA elements, one proximal and one distal to the LpS1 beta transcriptional start site, which bind to an ectoderm-enriched nuclear factor. In this report, we show that the ectoderm G-string factor binds to a G/C-rich region larger than the G-string itself and that the binding of the G-string factor requires sequences immediately downstream from the G-string. These downstream sequences are essential for full promoter activity. Two regions of 5'-flanking DNA are required for positive control of LpS1 beta, region I from base pairs -762 to -511, and region II, which includes the G/C-rich element, from base pairs -108 to -61. Region I also contains a mesenchyme cell repressor element. The results indicate that LpS1 beta expression is regulated positively in ectoderm cells and negatively in mesenchyme cells. Similar positive and negative control mechanisms regulate the expression of the related Spec genes of Strongylocentrotus purpuratus, but in this gene family the DNA elements are entirely different. We hypothesize that cis-regulatory elements are evolutionarily dynamic and that many different combinations of DNA elements are capable of given rise to aboral ectoderm-specific expression. PMID:8634141

  15. Effects of seawater acidification on gene expression: resolving broader-scale trends in sea urchins.

    PubMed

    Evans, Tyler G; Watson-Wynn, Priscilla

    2014-06-01

    Sea urchins are ecologically and economically important calcifying organisms threatened by acidification of the global ocean caused by anthropogenic CO2 emissions. Propelled by the sequencing of the purple sea urchin (Strongylocentrotus purpuratus) genome, profiling changes in gene expression during exposure to high pCO2 seawater has emerged as a powerful and increasingly common method to infer the response of urchins to ocean change. However, analyses of gene expression are sensitive to experimental methodology, and comparisons between studies of genes regulated by ocean acidification are most often made in the context of major caveats. Here we perform meta-analyses as a means of minimizing experimental discrepancies and resolving broader-scale trends regarding the effects of ocean acidification on gene expression in urchins. Analyses across eight studies and four urchin species largely support prevailing hypotheses about the impact of ocean acidification on marine calcifiers. The predominant expression pattern involved the down-regulation of genes within energy-producing pathways, a clear indication of metabolic depression. Genes with functions in ion transport were significantly over-represented and are most plausibly contributing to intracellular pH regulation. Expression profiles provided extensive evidence for an impact on biomineralization, epitomized by the down-regulation of seven spicule matrix proteins. In contrast, expression profiles provided limited evidence for CO2-mediated developmental delay or induction of a cellular stress response. Congruence between studies of gene expression and the ocean acidification literature in general validates the accuracy of gene expression in predicting the consequences of ocean change and justifies its continued use in future studies. PMID:25070868

  16. SM50 repeat-polypeptides self-assemble into discrete matrix subunits and promote appositional calcium carbonate crystal growth during sea urchin tooth biomineralization.

    PubMed

    Mao, Yelin; Satchell, Paul G; Luan, Xianghong; Diekwisch, Thomas G H

    2016-01-01

    The two major proteins involved in vertebrate enamel formation and echinoderm sea urchin tooth biomineralization, amelogenin and SM50, are both characterized by elongated polyproline repeat domains in the center of the macromolecule. To determine the role of polyproline repeat polypeptides in basal deuterostome biomineralization, we have mapped the localization of SM50 as it relates to crystal growth, conducted self-assembly studies of SM50 repeat polypeptides, and examined their effect on calcium carbonate and apatite crystal growth. Electron micrographs of the growth zone of Strongylocentrotus purpuratus sea urchin teeth documented a series of successive events from intravesicular mineral nucleation to mineral deposition at the interface between tooth surface and odontoblast syncytium. Using immunohistochemistry, SM50 was detected within the cytoplasm of cells associated with the developing tooth mineral, at the mineral secreting front, and adjacent to initial mineral deposits, but not in muscles and ligaments. Polypeptides derived from the SM50 polyproline alternating hexa- and hepta-peptide repeat region (SM50P6P7) formed highly discrete, donut-shaped self-assembly patterns. In calcium carbonate crystal growth studies, SM50P6P7 repeat peptides triggered the growth of expansive networks of fused calcium carbonate crystals while in apatite growth studies, SM50P6P7 peptides facilitated the growth of needle-shaped and parallel arranged crystals resembling those found in developing vertebrate enamel. In comparison, SM50P6P7 surpassed the PXX24 polypeptide repeat region derived from the vertebrate enamel protein amelogenin in its ability to promote crystal nucleation and appositional crystal growth. Together, these studies establish the SM50P6P7 polyproline repeat region as a potent regulator in the protein-guided appositional crystal growth that occurs during continuous tooth mineralization and eruption. In addition, our studies highlight the role of species

  17. Experimental ocean acidification alters the allocation of metabolic energy

    PubMed Central

    Pan, T.-C. Francis; Applebaum, Scott L.; Manahan, Donal T.

    2015-01-01

    Energy is required to maintain physiological homeostasis in response to environmental change. Although responses to environmental stressors frequently are assumed to involve high metabolic costs, the biochemical bases of actual energy demands are rarely quantified. We studied the impact of a near-future scenario of ocean acidification [800 µatm partial pressure of CO2 (pCO2)] during the development and growth of an important model organism in developmental and environmental biology, the sea urchin Strongylocentrotus purpuratus. Size, metabolic rate, biochemical content, and gene expression were not different in larvae growing under control and seawater acidification treatments. Measurements limited to those levels of biological analysis did not reveal the biochemical mechanisms of response to ocean acidification that occurred at the cellular level. In vivo rates of protein synthesis and ion transport increased ∼50% under acidification. Importantly, the in vivo physiological increases in ion transport were not predicted from total enzyme activity or gene expression. Under acidification, the increased rates of protein synthesis and ion transport that were sustained in growing larvae collectively accounted for the majority of available ATP (84%). In contrast, embryos and prefeeding and unfed larvae in control treatments allocated on average only 40% of ATP to these same two processes. Understanding the biochemical strategies for accommodating increases in metabolic energy demand and their biological limitations can serve as a quantitative basis for assessing sublethal effects of global change. Variation in the ability to allocate ATP differentially among essential functions may be a key basis of resilience to ocean acidification and other compounding environmental stressors. PMID:25825763

  18. The Protein Precursors of Peptides That Affect the Mechanics of Connective Tissue and/or Muscle in the Echinoderm Apostichopus japonicus

    PubMed Central

    Elphick, Maurice R.

    2012-01-01

    Peptides that cause muscle relaxation or contraction or that modulate electrically-induced muscle contraction have been discovered in the sea cucumber Apostichopus japonicus (Phylum Echinodermata; Class Holothuroidea). By analysing transcriptome sequence data, here the protein precursors of six of these myoactive peptides (the SALMFamides Sticho-MFamide-1 and -2, NGIWYamide, stichopin, GN-19 and GLRFA) have been identified, providing novel insights on neuropeptide and endocrine-type signalling systems in echinoderms. The A. japonicus SALMFamide precursor comprises eight putative neuropeptides including both L-type and F-type SALMFamides, which contrasts with previous findings from the sea urchin Strongylocentrotus purpuratus where L-type and F-type SALMFamides are encoded by different genes. The NGIWYamide precursor contains five copies of NGIWYamide but, unlike other NG peptide-type neuropeptide precursors in deuterostomian invertebrates, the NGIWYamide precursor does not have a C-terminal neurophysin domain, indicating loss of this character in holothurians. NGIWYamide was originally discovered as a muscle contractant, but it also causes stiffening of mutable connective tissue in the body wall of A. japonicus, whilst holokinins (PLGYMFR and derivative peptides) cause softening of the body wall. However, the mechanisms by which these peptides affect the stiffness of body wall connective tissue are unknown. Interestingly, analysis of the A. japonicus transcriptome reveals that the only protein containing the holokinin sequence PLGYMFR is an alpha-5 type collagen. This suggests that proteolysis of collagen may generate peptides (holokinins) that affect body wall stiffness in sea cucumbers, providing a novel perspective on mechanisms of mutable connective tissue in echinoderms. PMID:22952987

  19. De novo assembly and analysis of tissue-specific transcriptomes revealed the tissue-specific genes and profile of immunity from Strongylocentrotus intermedius.

    PubMed

    Chen, Yadong; Chang, Yaqing; Wang, Xiuli; Qiu, Xuemei; Liu, Yang

    2015-10-01

    Strongylocentrotus intermedius is an important marine species in north China and Japan. Recent years, diseases are threating the sea urchin aquaculture industry seriously. To provide a genetic resource for S. intermedius as well as overview the immune-related genes of S. intermedius, we performed transcriptome sequencing of three cDNA libraries representing three tissues, coelomocytes, gut and peristomial membrane respectively. In total 138,421 contigs were assembled from all sequencing data. 96,764 contigs were annotated according to bioinformatics databases, including NT, nr, Swiss-Prot, KEGG, COG. 49,336 Contigs were annotated as CDS. In this study, we obtained 24,778 gene families from S. intermedius transcriptome. The gene expression analysis revealed that more genes were expressed in gut, more high expression level genes in coelomocytes when compared with other tissues. Specific expressed contigs in coelomocytes, gut, and peristomial membrane were 546, 1136, and 1012 respectively. Pathway analysis suggested 25, 17 and 36 potential specifically pathways may specific progressed in peristomial membrane, gut and coelomocytes respectively. Similarities and differences between S. intermedius and other echinoderms were analyzed. S. intermedius was more homology to Strongylocentrotus purpuratus than others sea urchin. Of 24,778 genes, 1074 genes are immune-related, immune genes were expressed with a higher level in coelomocytes than other tissues. Complement system may be the most important immune system in sea urchin. We also identified 2438 SSRs and 16,236 SNPs for S. intermedius. These results provide a transcriptome resource and foundation to study molecular mechanisms of sea urchin immune system. PMID:26253994

  20. Centrocins: isolation and characterization of novel dimeric antimicrobial peptides from the green sea urchin, Strongylocentrotus droebachiensis.

    PubMed

    Li, Chun; Haug, Tor; Moe, Morten K; Styrvold, Olaf B; Stensvåg, Klara

    2010-09-01

    As immune effector molecules, antimicrobial peptides (AMPs) play an important role in the invertebrate immune system. Here, we present two novel AMPs, named centrocins 1 (4.5kDa) and 2 (4.4kDa), purified from coelomocyte extracts of the green sea urchin, Strongylocentrotus droebachiensis. The native peptides are cationic and show potent activities against Gram-positive and Gram-negative bacteria. The centrocins have an intramolecular heterodimeric structure, containing a heavy chain (30 amino acids) and a light chain (12 amino acids). The cDNA encoding the peptides and genomic sequences were cloned and sequenced. One putative isoform (centrocin 1b) was identified and one intron was found in the genes coding for the centrocins. The full length protein sequence of centrocin 1 consists of 119 amino acids, whereas centrocin 2 consists of 118 amino acids which both include a preprosequence of 51 or 50 amino acids for centrocins 1 and 2, respectively, and an interchain of 24 amino acids between the heavy and light chain. The difference of molecular mass between the native centrocins and the deduced sequences from cDNA indicates that the native centrocins contain a post-translational brominated tryptophan. In addition, two amino acids at the C-terminal, Gly-Arg, were removed from the light chains during the post-translational processing. The separate peptide chains of centrocin 1 were synthesized and the heavy chain alone was shown to be sufficient for antimicrobial activity. The genome of the closely related species, the purple sea urchin (S. purpuratus), was shown to contain two putative proteins with high similarity to the centrocins. PMID:20438753

  1. Geometrical analysis of the structure in the western flank of the Chilean Andes at 34°15'S-34°30'S

    NASA Astrophysics Data System (ADS)

    Jara, P. P.; Charrier, R.; Arriagada, C.; Farias, M.

    2007-12-01

    We present a structural analysis of strongly deformed Cenozoic volcanic rocks of the western Principal Cordillera between 34°15'S and 34°30'S. Deformation has been related to the inversion of the Abanico Basin occurred between 22 and 16 Ma. The structural array is characterized by a syncline with a very steep western limb and a rather flat slightly folded axial zone. On both sides of this structure, layers dip in directions opposite to the syncline flanks, suggesting disruption of anticlines by faulting. The reconstructed geometry performed on 6 cross- sections indicates that: 1. Deformation occurred by "fault-propagation folding" with considerable fault displacement, and 2. The anticlines were transported by the faults cutting their crests along the axial plane, similar to the anticlinal breakthrough deformation modes. Based on this mechanism and the relations between the axial and cut off angles, we developed a kinematic model using geometric methods and the Trishear program. Results suggests that folding was controlled by two E-vergent faults (western, WF, and eastern, EF). The most important (WF) would have a cutoff angle 40°-60°, which is compatible with inverted normal faults. Indeed, the regional nature of shortening in this region has been interpreted as the result of basin inversion. The EF has a cutoff angle of about 20 degrees, which is compatible with a neoformed thrust and allowing its interpretation as a short cut. These structures have accommodated ca. 30% (2 to 3 km) of shortening in this region. Although our proposition at depth for the main fault geometry is based on the surface analysis, it is quite consistent with the structural array interpreted by seismicity and MT images. Moreover, this structure seems to be the upward prolongation of a W-dipping ramp structure, connecting the subduction zone with the tectonic front of the mountain belt, in a zone where the ramp flattens and forms a detachment at 15-20 km depth beneath the western edge of

  2. Searching Structural Control on the Evolution of the Reservoir Permeability in the Porphyry Copper-Molybdenum Rio Blanco - Los Bronces District: Structural Geology and Passive Seismic Tomography

    NASA Astrophysics Data System (ADS)

    Comte, D.; Carrizo, D.; Vela, I.; Silva, W.

    2013-05-01

    Understand main factors that controls the crustal permeability related to giant ore deposits is a crucial goal for improving metallogenic models. Subsurface data at distrital scales, remains a key factor to reach that goal, however use of regional geophysical methodologies are still extremely expensive and poorly developed on ore exploration. We present the results associated to distrital structural analyses, combined with passive seismic tomography in a case study in Central Andes. Tectono-magmatic evolution of the Rio Blanco-Los Bronces District (Porphyry Cu-Mo Deposit) involves a complex interaction of different processes along the time which includes: Pre-Mesozoic structural inheritance, Oligo-Miocene extension, Mio-Pliocene orogenic shortening with the respective tectonic inversion associated to the uplift of the Andean western margin. Miocene volcano sedimentary units (Abanico Formation) and Pliocene syn-orogenic volcanic units form the deformed host distrital structure. These rocks are part of a trench-linked Andean deformation belt that accommodates EW shortening by NS- strike faults and NS-axes fault related folds. This structural belt controls the uplift of the western margin of the Andes by tectonic inversion of Miocene rift-type structural geometries. The reservoir structure is interpreted by the emplacement of a batholith in the core of the transfer zone between two NS-strike reverse faults, where the ore bodies are located in the center of an anticline general structure showing NW-SE mineralization trend. In turn, NE trending faults dislocate ore bodies in the transfer zone. The reservoir structure is interpreted by the emplacement of a Mio-Pliocene syntectonic batholith in the middle of a transfer zone between two NS-strike reverse faults, where the ore bodies are located in the axis of the anticlinal general structure. Passive seismic tomography results suggest a NNW-strike zone formed by fractured or/and hydrated rocks. This zone is spatially

  3. Signs of adaptation to local pH conditions across an environmental mosaic in the California Current Ecosystem.

    PubMed

    Pespeni, M H; Chan, F; Menge, B A; Palumbi, S R

    2013-11-01

    Little is known about the potential for rapid evolution in natural populations in response to the high rate of contemporary climatic change. Organisms that have evolved in environments that experience high variability across space and time are of particular interest as they may harbor genetic variation that can facilitate evolutionary response to changing conditions. Here we review what is known about genetic capacity for adaptation in the purple sea urchin, Strongylocentrotus purpuratus, a species that has evolved in the upwelling ecosystem of the Northeast Pacific Ocean. We also present new results testing for adaptation to local pH conditions in six populations from Oregon to southern California. We integrate data on 19,493 genetic polymorphisms with data on local pH conditions. We find correlations between allele frequency and rank average time spent at pH <7.8 in 318 single-nucleotide polymorphisms in 275 genes. Two of the genes most correlated with local pH are a protein associated with the cytoskeleton and a proton pump, with functional roles in maintenance of cell volume and with internal regulation of pH, respectively. Across all loci tested, high correlations with local pH were concentrated in genes related to transport of ions, biomineralization, lipid metabolism, and cell-cell adhesion, functional pathways important for maintaining homeostasis at low pH. We identify a set of seven genes as top candidates for rapid evolutionary response to acidification of the ocean. In these genes, the putative low-pH-adapted allele, based on allele frequencies in natural populations, rapidly increases in frequency in purple sea urchin larvae raised at low pH. We also found that populations from localities with high pH show a greater change in allele frequency toward putative low-pH-adapted alleles under experimental acidification, compared with low-pH populations, suggesting that both natural and artificial selection favor the same alleles for response to low pH. These

  4. Juvenile skeletogenesis in anciently diverged sea urchin clades.

    PubMed

    Gao, Feng; Thompson, Jeffrey R; Petsios, Elizabeth; Erkenbrack, Eric; Moats, Rex A; Bottjer, David J; Davidson, Eric H

    2015-04-01

    Mechanistic understanding of evolutionary divergence in animal body plans devolves from analysis of those developmental processes that, in forms descendant from a common ancestor, are responsible for their morphological differences. The last common ancestor of the two extant subclasses of sea urchins, i.e., euechinoids and cidaroids, existed well before the Permian/Triassic extinction (252 mya). Subsequent evolutionary divergence of these clades offers in principle a rare opportunity to solve the developmental regulatory events underlying a defined evolutionary divergence process. Thus (i) there is an excellent and fairly dense (if yet incompletely analyzed) fossil record; (ii) cladistically confined features of the skeletal structures of modern euechinoid and cidaroid sea urchins are preserved in fossils of ancestral forms; (iii) euechinoids and cidaroids are among current laboratory model systems in molecular developmental biology (here Strongylocentrotus purpuratus [Sp] and Eucidaris tribuloides [Et]); (iv) skeletogenic specification in sea urchins is uncommonly well understood at the causal level of interactions of regulatory genes with one another, and with known skeletogenic effector genes, providing a ready arsenal of available molecular tools. Here we focus on differences in test and perignathic girdle skeletal morphology that distinguish all modern euechinoid from all modern cidaroid sea urchins. We demonstrate distinct canonical test and girdle morphologies in juveniles of both species by use of SEM and X-ray microtomography. Among the sharply distinct morphological features of these clades are the internal skeletal structures of the perignathic girdle to which attach homologous muscles utilized for retraction and protraction of Aristotles׳ lantern and its teeth. We demonstrate that these structures develop de novo between one and four weeks after metamorphosis. In order to study the underlying developmental processes, a method of section whole mount in

  5. A provisional regulatory gene network for specification of endomesoderm in the sea urchin embryo

    NASA Technical Reports Server (NTRS)

    Davidson, Eric H.; Rast, Jonathan P.; Oliveri, Paola; Ransick, Andrew; Calestani, Cristina; Yuh, Chiou-Hwa; Minokawa, Takuya; Amore, Gabriele; Hinman, Veronica; Arenas-Mena, Cesar; Otim, Ochan; Brown, C. Titus; Livi, Carolina B.; Lee, Pei Yun; Revilla, Roger; Schilstra, Maria J.; Clarke, Peter J C.; Rust, Alistair G.; Pan, Zhengjun; Arnone, Maria I.; Rowen, Lee; Cameron, R. Andrew; McClay, David R.; Hood, Leroy; Bolouri, Hamid

    2002-01-01

    We present the current form of a provisional DNA sequence-based regulatory gene network that explains in outline how endomesodermal specification in the sea urchin embryo is controlled. The model of the network is in a continuous process of revision and growth as new genes are added and new experimental results become available; see http://www.its.caltech.edu/mirsky/endomeso.htm (End-mes Gene Network Update) for the latest version. The network contains over 40 genes at present, many newly uncovered in the course of this work, and most encoding DNA-binding transcriptional regulatory factors. The architecture of the network was approached initially by construction of a logic model that integrated the extensive experimental evidence now available on endomesoderm specification. The internal linkages between genes in the network have been determined functionally, by measurement of the effects of regulatory perturbations on the expression of all relevant genes in the network. Five kinds of perturbation have been applied: (1) use of morpholino antisense oligonucleotides targeted to many of the key regulatory genes in the network; (2) transformation of other regulatory factors into dominant repressors by construction of Engrailed repressor domain fusions; (3) ectopic expression of given regulatory factors, from genetic expression constructs and from injected mRNAs; (4) blockade of the beta-catenin/Tcf pathway by introduction of mRNA encoding the intracellular domain of cadherin; and (5) blockade of the Notch signaling pathway by introduction of mRNA encoding the extracellular domain of the Notch receptor. The network model predicts the cis-regulatory inputs that link each gene into the network. Therefore, its architecture is testable by cis-regulatory analysis. Strongylocentrotus purpuratus and Lytechinus variegatus genomic BAC recombinants that include a large number of the genes in the network have been sequenced and annotated. Tests of the cis-regulatory predictions of

  6. First molluscan TNFR homologue in Zhikong scallop: molecular characterization and expression analysis.

    PubMed

    Li, Ling; Qiu, Limei; Song, Linsheng; Song, Xiaoyan; Zhao, Jianmin; Wang, Lingling; Mu, Changkao; Zhang, Huan

    2009-11-01

    Tumor necrosis factor receptors (TNFRs) are a superfamily of proteins characterized by the unique cysteine-rich domain (CRD) and their important roles in diverse physiological and pathological events such as inflammation, apoptosis, autoimmunity and organogenesis. The first member of the molluscan TNFR family, designated as CfTNFR, was identified from Zhikong scallop Chlamys farreri by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of CfTNFR was of 1334 bp, consisting of a 5' UTR of 17 bp, a 3' UTR of 69 bp with a poly (A) tail, and an open reading frame (ORF) of 1248 bp encoding a polypeptide of 415 amino acids with a theoretical isoelectric point of 8.33 and predicted molecular weight of 47.07 kDa. There were a signal peptide, a CRD, a transmembrane region and a death domain in the deduced amino acid sequence of CfTNFR, suggesting that it was a typical type I membrane protein. The high identities (22-40%) of CfTNFR with other TNFR superfamily members indicated that CfTNFR should be a member of TNFR superfamily, and moreover, it should be the first death domain-containing TNFR found in invertebrates. Phylogenetic analysis revealed that CfTNFR was closely related to TNFR-like proteins from Strongylocentrotus purpuratus, Drosophila melanogaster and Ciona intestinalis, and they formed a separate branch apart from vertebrate TNFRs. The spatial expression of CfTNFR transcripts in healthy and bacteria challenged scallops was examined by quantitative real-time PCR. CfTNFR transcripts could be detected in all tested tissues, including haemocytes, gonad, gill, mantle and hepatopancreas, and significantly up-regulated in the tissues of gonad, gill, mantle and hepatopancreas after Listonella anguillarum challenge, indicating that CfTNFR was constitutive and inducible acute-phase protein involved in immune defence. The present results suggested the existence of the TNFR-like molecules and TNF-TNFR system in low

  7. Innate immune complexity in the purple sea urchin: diversity of the sp185/333 system.

    PubMed

    Smith, L Courtney

    2012-01-01

    The California purple sea urchin, Strongylocentrotus purpuratus, is a long-lived echinoderm with a complex and sophisticated innate immune system. There are several large gene families that function in immunity in this species including the Sp185/333 gene family that has ∼50 (±10) members. The family shows intriguing sequence diversity and encodes a broad array of diverse yet similar proteins. The genes have two exons of which the second encodes the mature protein and has repeats and blocks of sequence called elements. Mosaics of element patterns plus single nucleotide polymorphisms-based variants of the elements result in significant sequence diversity among the genes yet maintains similar structure among the members of the family. Sequence of a bacterial artificial chromosome insert shows a cluster of six, tightly linked Sp185/333 genes that are flanked by GA microsatellites. The sequences between the GA microsatellites in which the Sp185/333 genes and flanking regions are located, are much more similar to each other than are the sequences outside the microsatellites suggesting processes such as gene conversion, recombination, or duplication. However, close linkage does not correspond with greater sequence similarity compared to randomly cloned and sequenced genes that are unlikely to be linked. There are three segmental duplications that are bounded by GAT microsatellites and include three almost identical genes plus flanking regions. RNA editing is detectible throughout the mRNAs based on comparisons to the genes, which, in combination with putative post-translational modifications to the proteins, results in broad arrays of Sp185/333 proteins that differ among individuals. The mature proteins have an N-terminal glycine-rich region, a central RGD motif, and a C-terminal histidine-rich region. The Sp185/333 proteins are localized to the cell surface and are found within vesicles in subsets of polygonal and small phagocytes. The coelomocyte proteome shows full

  8. Phylogenomics of strongylocentrotid sea urchins

    PubMed Central

    2013-01-01

    Background Strongylocentrotid sea urchins have a long tradition as model organisms for studying many fundamental processes in biology including fertilization, embryology, development and genome regulation but the phylogenetic relationships of the group remain largely unresolved. Although the differing isolating mechanisms of vicariance and rapidly evolving gamete recognition proteins have been proposed, a stable and robust phylogeny is unavailable. Results We used a phylogenomic approach with mitochondrial and nuclear genes taking advantage of the whole-genome sequencing of nine species in the group to establish a stable (i.e. concordance in tree topology among multiple lies of evidence) and robust (i.e. high nodal support) phylogenetic hypothesis for the family Strongylocentrotidae. We generated eight draft mitochondrial genome assemblies and obtained 13 complete mitochondrial genes for each species. Consistent with previous studies, mitochondrial sequences failed to provide a reliable phylogeny. In contrast, we obtained a very well-supported phylogeny from 2301 nuclear genes without evidence of positive Darwinian selection both from the majority of most-likely gene trees and the concatenated fourfold degenerate sites: ((P. depressus, (M. nudus, M. franciscanus), (H. pulcherrimus, (S. purpuratus, (S. fragilis, (S. pallidus, (S. droebachiensis, S. intermedius)). This phylogeny was consistent with a single invasion of deep-water environments followed by a holarctic expansion by Strongylocentrotus. Divergence times for each species estimated with reference to the divergence times between the two major clades of the group suggest a correspondence in the timing with the opening of the Bering Strait and the invasion of the holarctic regions. Conclusions Nuclear genome data contains phylogenetic signal informative for understanding the evolutionary history of this group. However, mitochondrial genome data does not. Vicariance can explain major patterns observed in the

  9. Early-Middle Cenozoic Andean mammal faunas: Integrated analyses of biochronology, geochronology, and paleoecology (Invited)

    NASA Astrophysics Data System (ADS)

    Flynn, J. J.

    2010-12-01

    For almost two centuries, understanding of the South American Cenozoic terrestrial biota was derived largely from the extensive but gap-riddled record from Patagonia and nearby lowland, high-latitude sites. But discoveries and analyses of Andean and tropical fossil mammal assemblages have increased substantially in recent years, and integrating these new paleontological data with those typically used in geochronologic and tectonic studies can yield new or deeper insights into the timing, origin, and magnitude of biotic responses to environmental, climatic and other physical changes, including the influences of regional (e.g., tectonism) versus global (e.g., climate change) events. More than two decades of collaborative research with R. Charrier (U. Chile), A. Wyss and P. Gans (UC-Santa Barbara), D. Croft (Case Western), the National Museum of Chile, and other investigators in the Main Range of the Chilean Andes is creating one of the premier archives of early-middle Cenozoic terrestrial mammal fossils. The active margin setting and thick volcaniclastic sequences accumulating in Andean extensional basins foster preservation of a unique record of mammalian evolution, and development of a more precise and reliable terrestrial geochronology integrating biochronology, magnetostratigraphy and high-precision radioisotopic dating, including the first calibration for some South American Land Mammal “Ages” (SALMAs). Intensive work within the Andes of Chile (particularly the Abanico Fm. and its equivalents, from 33°-38°30’S) has yielded more than 3,000 specimens from > 2 dozen sets of localities, spanning some 30° of latitude and ranging in age from at least 40 to 10 Ma (late Eocene to late Miocene). Exemplar “case-studies” illustrate how these new fossils and dates provide key data for understanding mammalian evolution and paleoecology, documenting faunal change through time (during periods of profound environmental and biotic restructuring), assessing

  10. The Eastern delta-fan deposits on the Granada Basin as tectonic indicators of the Sierra Nevada uplift (Betic Cordillera, South Spain)

    NASA Astrophysics Data System (ADS)

    Roldán, Francisco Javier; Azañon, Jose Miguel; Mateos, Rosa Maria

    2014-05-01

    Sierra Nevada uplift and with original NE-SW normal left-lateral fault system associated with a compatible ESE-WNW normal fault system. The asymmetrical morphologies of the two delta-fan bodies confirm this configuration as tectonic induced deposits. References Braga, J.C., Martín, J.M. and Alcalá, B. (1990). Coral reefs in coarse-terrigenous sedimentary environments (Upper Tortonian, Granada Basin, southern Spain). Sedimentary Geology, 66, pp 135-150. Clark, S.J.P.and Dempster, T.J. (2009). The record of tectonic denudation and erosion in an emerging orogen: an apatite fission-track study of the Sierra Nevada, southern Spain. Journal of the Geological Society, London, Vol. 166, pp. 87-100. Dabrio, C.J., Fernández, J., Peña, J., Ruíz-Bustos, A. y Sanz de Galdeano, C. (1978). Rasgos sedimentarios de los conglomerados miocénicos del borde noreste de la Depresión de Granada. Estudios Geológicos, 34, pp 89-97. García-García, F., Fernández, J., Viseras, C. and Soria, J.M. (2006). Architecture and sedimentary facies evolution in a delta stack controlled by fault growth (Betic Cordillera, southern Spain, late Tortonian). Sedimentary Geology, 185, pp. 79-92. García-García, F., Viseras, C. and Fernández, J. (1999). Organización secuencial de abanicos deltaicos controlados por la tectónica (Tortoniense superior, cuenca de Granada, Cordillera Bética). Rev. Soc. Geol. España, 12 (2): 199-208.

  11. Evolutionary Dynamics of the wnt Gene Family: A Lophotrochozoan Perspective

    PubMed Central

    Cho, Sung-Jin; Vallès, Yvonne; Giani, Vincent C.; Seaver, Elaine C.; Weisblat, David A.

    2010-01-01

    The wnt gene family encodes a set of secreted glycoproteins involved in key developmental processes, including cell fate specification and regulation of posterior growth (Cadigan KM, Nusse R. 1997. Wnt signaling: a common theme in animal development. Genes Dev. 11:3286–3305.; Martin BL, Kimelman D. 2009. Wnt signaling and the evolution of embryonic posterior development. Curr Biol. 19:R215–R219.). As for many other gene families, evidence for expansion and/or contraction of the wnt family is available from deuterostomes (e.g., echinoderms and vertebrates [Nusse R, Varmus HE. 1992. Wnt genes. Cell. 69:1073–1087.; Schubert M, Holland LZ, Holland ND, Jacobs DK. 2000. A phylogenetic tree of the Wnt genes based on all available full-length sequences, including five from the cephalochordate amphioxus. Mol Biol Evol. 17:1896–1903.; Croce JC, Wu SY, Byrum C, Xu R, Duloquin L, Wikramanayake AH, Gache C, McClay DR. 2006. A genome-wide survey of the evolutionarily conserved Wnt pathways in the sea urchin Strongylocentrotus purpuratus. Dev Biol. 300:121–131.]) and ecdysozoans (e.g., arthropods and nematodes [Eisenmann DM. 2005. Wnt signaling. WormBook. 1–17.; Bolognesi R, Farzana L, Fischer TD, Brown SJ. 2008. Multiple Wnt genes are required for segmentation in the short-germ embryo of Tribolium castaneum. Curr Biol. 18:1624–1629.]), but little is known from the third major bilaterian group, the lophotrochozoans (e.g., mollusks and annelids [Prud'homme B, Lartillot N, Balavoine G, Adoutte A, Vervoort M. 2002. Phylogenetic analysis of the Wnt gene family. Insights from lophotrochozoan members. Curr Biol. 12:1395.]). To obtain a more comprehensive scenario of the evolutionary dynamics of this gene family, we exhaustively mined wnt gene sequences from the whole genome assemblies of a mollusk (Lottia gigantea) and two annelids (Capitella teleta and Helobdella robusta) and examined them by phylogenetic, genetic linkage, intron–exon structure, and embryonic