Sea urchin embryos build their endoskeleton following a well defined time- and space-dependent program, that is not yet fully understood despite the many studies on this classical renowned embryological model. The availability of the sea urchin genome published in 2006[1] has expanded the possibilities to investigate the skeletogenic program at the gene level, with an obvious positive impact on the knowledge of the biomineralization process. In the sea urchin embryo, the biomineral is produced by a specialized population of mesodermal cells, arranging themselves in a definite pattern at the gastrula stage, and it is deposited in the extracellular spaces in between cell fused filopodia[2]. The capability of mesodermal cells to produce the necessary skeleton matrix proteins which will constitute, together with magnesian calcite, the embryonic spicules is activated by growth factors deriving from the nearby embryonic ectodermal cells, as we showed for the first time more than ten years ago[3]. Since then, many of the crucial steps of this complex activation cascade, from ectoderm cells to embryonic spicules, have been elucidated. Research is also directed to the understanding of environmental impacts on biomineral formation as a consequence of embryo exposure to stressors, including toxic metals [4-9] and ionizing radiations [9-15]. Our studies always showed, although with peculiarities dependent on the stressor used, major defects in skeleton elongation and patterning that we correlated with the modulation of specific biomarkers, at the transductional, translational and post-translational levels. In recent years, the approaches used in our laboratory to dissect the molecular bases of biomineralization included: identification of the skelegenesis-related cDNAs, cloning and characterization of their sequences, study of developmentally-relevant spatial and temporal gene expression, phylogenetic analysis, recombinant proteins and specific antibodies production, functional biological assays. To date we developed a molecular toolset of cDNAs, recombinant proteins and specific antibodies, to be used in molecular, biochemical, cellular and physiological studies, with which we expect to contribute to the understanding of sea urchin embryo biomineralization [16-19]., as well as adult exploitantion[9-15]. . In addition, we are developing new biotechnological approaches for the treatment of biomineral-associated pathologies in humans by the production and application of new recombinant proteins in homologous and heterologous systems. References [1] Sea Urchin Genome Sequencing Consortium (2006) The genome of the sea urchin Strongylocentrotus purpuratus. Science 314, 941-952. [2] Matranga V, Bonaventura R, Costa C, Karakostis K, Pinsino A, Russo R, Zito F (2011). Echinoderms as blueprints for biocalcification: regulation of skeletogenic genes and matrices. Prog Mol Subcell Biol 52:225-248. [3] Zito, F., Costa, C., Sciarrino, S., Poma, V., Russo, R., Angerer, L. M., and Matranga, V. (2003). Expression of univin, a TGF-beta growth factor, requires ectoderm-ECM interaction and promotes skeletal growth in the sea urchin embryo. Dev. Biol. 264, 217-227 [4] Russo R, Bonaventura R, Zito F, Schröder HC, Müller I, Müller WE, Matranga V (2003) Stress to cadmium monitored by metallothionein gene induction in Paracentrotus lividus embryos. Cell Stress Chaperones 8: 232-241 [5] Roccheri M.C., Agnello M., Bonaventura R., Matranga V. 2004. Cadmium induces the expression of specific stress proteins in sea urchin embryos. Biochemical and Biophysical Research Communications, 321:80-87. [6] Filosto S., Roccheri M.C., Bonaventura R. & Matranga V. 2008. Environmentally relevant cadmium concentrations affect development and induce apoptosis of Paracentrotus lividus larvae cultured in vitro. Cell Biology and Toxicology, 24:603-610. [7] Pinsino A., Matranga V., Trinchella F. & Roccheri M.C. 2010. Sea urchin embryos as an in vivo model for the assessment of manganese toxicity: developmental and stress response effects. Ecotoxicology, 19:555-562. [8] Pinsino A, Roccheri MC, Costa C, Matranga V (2011) Manganese interferes with calcium, perturbs ERK signaling, and produces embryos with no skeleton. Toxicol Sci. 123:217-30. [9] Pinsino A., Roccheri M.C., Matranga V. (2013) Manganese overload affects p38 MAPK phosphorylation and metalloproteinase activity during sea urchin embryonic development. Marine Environmental Research, submitted [10] Bonaventura R., Poma V., Costa C., Matranga V. 2005. UVB radiation prevents skeleton growth and stimulates the expression of stress markers in sea urchin embryos. Biochemical and Biophysical Research Communications, 328:150-157. [11] Bonaventura R., Poma V., Russo R., Zito F., Matranga V. 2006. Effects of UV-B radiation on the development and hsp70 expression in sea urchin cleavage embryos. Marine Biology, 149: 79-86. Erratum in: Mar Biol (2007) 150: 1051 [12] Russo R, Zito F, Costa C, Bonaventura R, Matranga V (2010) Transcriptional increase and misexpression of 14-3-3 epsilon in sea urchin embryos exposed to UV-B. Cell Stress Chaperones. 15: 993-1001. [13] Russo R., Bonaventura R., Matranga V (2013) Early and late responsive genes affected by UVB radiation in sea urchin embryos. Marine Environmental Research, submitted [14] Matranga V, Zito F, Costa C, Bonaventura R, Giarrusso S & Celi F (2010) Embryonic development and skeletogenic gene expression affected by X-rays in the Mediterranean sea urchin Paracentrotus lividus. Ecotoxicology, 19:530-537. [15] Bonaventura R, Zito F, Costa C, Giarrusso S, Celi F, Matranga V (2011) Stress response gene activation protects sea urchin embryos exposed to X-rays. Cell Stress Chaperones 16: 681-687. [16] Costa C., Karakostis K., Zito F. & Matranga V. 2012. Phylogenetic analysis and expression patterns of p16 and p19 in Paracentrotus lividus embryos. Development Genes and Evolution, 222:245-51. [17] Karakostis K., Costa C., Zito F., Schröder H.C., Müller W.E.G., Matranga V (2013) Molecular characterization and biological activities of a newly identified galectin-8 from P. lividus sea urchin embryo. manuscript in preparation. [18] Karakostis K, Costa C, Brümmer F, Matranga V (2013) Characterization of a newly identified carbonic anhydrase from P. lividus involved in biomineralization. manuscript in preparation. [19] Karakostis K., Guichard N., Matranga V., Marin F (2013) Proteomic analysis of biomineralization proteins of the organic matrix from the adult test of the sea urchin Paracentrotus lividus. manuscript in preparation.
The sea urchin model and its use in biomineralization studies and applications.
V Matranga;R Bonaventura;C Costa;A Pinsino;R Russo;F Zito
2013
Abstract
Sea urchin embryos build their endoskeleton following a well defined time- and space-dependent program, that is not yet fully understood despite the many studies on this classical renowned embryological model. The availability of the sea urchin genome published in 2006[1] has expanded the possibilities to investigate the skeletogenic program at the gene level, with an obvious positive impact on the knowledge of the biomineralization process. In the sea urchin embryo, the biomineral is produced by a specialized population of mesodermal cells, arranging themselves in a definite pattern at the gastrula stage, and it is deposited in the extracellular spaces in between cell fused filopodia[2]. The capability of mesodermal cells to produce the necessary skeleton matrix proteins which will constitute, together with magnesian calcite, the embryonic spicules is activated by growth factors deriving from the nearby embryonic ectodermal cells, as we showed for the first time more than ten years ago[3]. Since then, many of the crucial steps of this complex activation cascade, from ectoderm cells to embryonic spicules, have been elucidated. Research is also directed to the understanding of environmental impacts on biomineral formation as a consequence of embryo exposure to stressors, including toxic metals [4-9] and ionizing radiations [9-15]. Our studies always showed, although with peculiarities dependent on the stressor used, major defects in skeleton elongation and patterning that we correlated with the modulation of specific biomarkers, at the transductional, translational and post-translational levels. In recent years, the approaches used in our laboratory to dissect the molecular bases of biomineralization included: identification of the skelegenesis-related cDNAs, cloning and characterization of their sequences, study of developmentally-relevant spatial and temporal gene expression, phylogenetic analysis, recombinant proteins and specific antibodies production, functional biological assays. To date we developed a molecular toolset of cDNAs, recombinant proteins and specific antibodies, to be used in molecular, biochemical, cellular and physiological studies, with which we expect to contribute to the understanding of sea urchin embryo biomineralization [16-19]., as well as adult exploitantion[9-15]. . In addition, we are developing new biotechnological approaches for the treatment of biomineral-associated pathologies in humans by the production and application of new recombinant proteins in homologous and heterologous systems. References [1] Sea Urchin Genome Sequencing Consortium (2006) The genome of the sea urchin Strongylocentrotus purpuratus. Science 314, 941-952. [2] Matranga V, Bonaventura R, Costa C, Karakostis K, Pinsino A, Russo R, Zito F (2011). Echinoderms as blueprints for biocalcification: regulation of skeletogenic genes and matrices. Prog Mol Subcell Biol 52:225-248. [3] Zito, F., Costa, C., Sciarrino, S., Poma, V., Russo, R., Angerer, L. M., and Matranga, V. (2003). Expression of univin, a TGF-beta growth factor, requires ectoderm-ECM interaction and promotes skeletal growth in the sea urchin embryo. Dev. Biol. 264, 217-227 [4] Russo R, Bonaventura R, Zito F, Schröder HC, Müller I, Müller WE, Matranga V (2003) Stress to cadmium monitored by metallothionein gene induction in Paracentrotus lividus embryos. Cell Stress Chaperones 8: 232-241 [5] Roccheri M.C., Agnello M., Bonaventura R., Matranga V. 2004. Cadmium induces the expression of specific stress proteins in sea urchin embryos. Biochemical and Biophysical Research Communications, 321:80-87. [6] Filosto S., Roccheri M.C., Bonaventura R. & Matranga V. 2008. Environmentally relevant cadmium concentrations affect development and induce apoptosis of Paracentrotus lividus larvae cultured in vitro. Cell Biology and Toxicology, 24:603-610. [7] Pinsino A., Matranga V., Trinchella F. & Roccheri M.C. 2010. Sea urchin embryos as an in vivo model for the assessment of manganese toxicity: developmental and stress response effects. Ecotoxicology, 19:555-562. [8] Pinsino A, Roccheri MC, Costa C, Matranga V (2011) Manganese interferes with calcium, perturbs ERK signaling, and produces embryos with no skeleton. Toxicol Sci. 123:217-30. [9] Pinsino A., Roccheri M.C., Matranga V. (2013) Manganese overload affects p38 MAPK phosphorylation and metalloproteinase activity during sea urchin embryonic development. Marine Environmental Research, submitted [10] Bonaventura R., Poma V., Costa C., Matranga V. 2005. UVB radiation prevents skeleton growth and stimulates the expression of stress markers in sea urchin embryos. Biochemical and Biophysical Research Communications, 328:150-157. [11] Bonaventura R., Poma V., Russo R., Zito F., Matranga V. 2006. Effects of UV-B radiation on the development and hsp70 expression in sea urchin cleavage embryos. Marine Biology, 149: 79-86. Erratum in: Mar Biol (2007) 150: 1051 [12] Russo R, Zito F, Costa C, Bonaventura R, Matranga V (2010) Transcriptional increase and misexpression of 14-3-3 epsilon in sea urchin embryos exposed to UV-B. Cell Stress Chaperones. 15: 993-1001. [13] Russo R., Bonaventura R., Matranga V (2013) Early and late responsive genes affected by UVB radiation in sea urchin embryos. Marine Environmental Research, submitted [14] Matranga V, Zito F, Costa C, Bonaventura R, Giarrusso S & Celi F (2010) Embryonic development and skeletogenic gene expression affected by X-rays in the Mediterranean sea urchin Paracentrotus lividus. Ecotoxicology, 19:530-537. [15] Bonaventura R, Zito F, Costa C, Giarrusso S, Celi F, Matranga V (2011) Stress response gene activation protects sea urchin embryos exposed to X-rays. Cell Stress Chaperones 16: 681-687. [16] Costa C., Karakostis K., Zito F. & Matranga V. 2012. Phylogenetic analysis and expression patterns of p16 and p19 in Paracentrotus lividus embryos. Development Genes and Evolution, 222:245-51. [17] Karakostis K., Costa C., Zito F., Schröder H.C., Müller W.E.G., Matranga V (2013) Molecular characterization and biological activities of a newly identified galectin-8 from P. lividus sea urchin embryo. manuscript in preparation. [18] Karakostis K, Costa C, Brümmer F, Matranga V (2013) Characterization of a newly identified carbonic anhydrase from P. lividus involved in biomineralization. manuscript in preparation. [19] Karakostis K., Guichard N., Matranga V., Marin F (2013) Proteomic analysis of biomineralization proteins of the organic matrix from the adult test of the sea urchin Paracentrotus lividus. manuscript in preparation.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
