The Experts below are selected from a list of 300 Experts worldwide ranked by ideXlab platform
Juliana Mozer Sciani - One of the best experts on this subject based on the ideXlab platform.
-
preliminary molecular characterization of a proinflammatory and nociceptive molecule from the Echinometra Lucunter spines extracts
Journal of Venomous Animals and Toxins Including Tropical Diseases, 2017Co-Authors: Juliana Mozer Sciani, Bianca Cestari Zychar, Luis Roberto De Camargo Goncalves, Thiago De Oliveira Nogueira, Renata Giorgi, Daniel C PimentaAbstract:Sea urchins are animals commonly found on the Brazilian shoreline, being Echinometra Lucunter the most abundant species. Accidents caused by E. Lucunter have been reported as one of the most frequent in Brazil, and are characterized by intense pain and inflammation, consequence of spine puncture in the skin. In order to characterize such toxic effects, we isolated one molecule that caused inflammatory and nociceptive effects. E. Lucunter specimens were collected without gender distinction. Spines were removed and molecules were extracted, fractionated by RP-HPLC and assayed for inflammatory and nociceptive activity, in a biological-driven fractionation way, until the obtainment of one active molecule and its subsequent analysis by mass spectrometry (MS and MS/MS). For inflammation, intravital microscopy was performed on the mouse cremaster muscle, in order to evaluate rolled, adherent and migrating leukocytes. Paw edema was also evaluated. For the nociceptive activity, the paw pressure test was performed in rats. One molecule could be isolated and related to the inflammatory and nociceptive activity. Regarding inflammation, increase in adherent and migrating cells was observed in the cremaster muscle after the administration of the molecule. Corroborating the inflammatory response, paw edema was also observed, although only in 20% of controls and 20 min after injection. Additionally, this molecule was able to decrease significantly the pain threshold, characterizing hyperalgesia. This molecule was analyzed by mass spectrometry, and according to the exact molecular mass, isotopic distribution and fragmentation profile, it was possible to propose the molecular formula C29H48N3O10. One isolated molecule from the spine extract of E. Lucunter is able to elicit inflammation and hypernociception in animal models, which is in agreement with the effects observed in sea urchin accidents.
-
Initial peptidomic profiling of Brazilian sea urchins: Arbacia lixula, Lytechinus variegatus and Echinometra Lucunter
Journal of Venomous Animals and Toxins including Tropical Diseases, 2016Co-Authors: Juliana Mozer Sciani, Andrews Krupinski Emerenciano, José Roberto Machado Cunha Da Silva, Daniel Carvalho PimentaAbstract:Background Sea urchins can be found throughout the Brazilian coast and are reported to be one of the major causes of marine accidents on the shoreline. Although not lethal, these accidents are reported to be extremely painful. In order to understand the toxinology of the Brazilian urchins, a peptidomic approach was performed aiming to characterize the naturally occurring peptides in both the coelomic fluid and the spine. Methods Animals were collected without gender distinction and samples of the coelomic fluid and spines extracted were analyzed by RP-HPLC and mass spectrometry for peptide de novo sequencing. Results Several peptides were identified either in the coelomic fluid or the spine extract (except for E. Lucunter ). The peptide sequences were aligned with public deposited sequences and possible functions were inferred. Moreover, some peptides can be cryptides, since their sequences were identified within functional proteins, for example thymosin from Strongylocentrotus purpuratus. Conclusions Although preliminary, the peptidomic approach presented here reports, for the first time, the abundance of novel biological molecules derived from these animals. The discovery of such molecules may be of potential biotechnological application, as described for other organisms; nevertheless, further studies are required.
-
initial peptidomic profiling of brazilian sea urchins arbacia lixula lytechinus variegatus and Echinometra Lucunter
Journal of Venomous Animals and Toxins Including Tropical Diseases, 2016Co-Authors: Juliana Mozer Sciani, Jose Roberto Machado Cunha Da Silva, Andrews Krupinski Emerenciano, Daniel C PimentaAbstract:Background Sea urchins can be found throughout the Brazilian coast and are reported to be one of the major causes of marine accidents on the shoreline. Although not lethal, these accidents are reported to be extremely painful. In order to understand the toxinology of the Brazilian urchins, a peptidomic approach was performed aiming to characterize the naturally occurring peptides in both the coelomic fluid and the spine.
-
echinometrin a novel mast cell degranulating peptide from the coelomic liquid of Echinometra Lucunter sea urchin
Peptides, 2014Co-Authors: Juliana Mozer Sciani, Bianca Cestari Zychar, Luis Roberto De Camargo Goncalves, Thiago De Oliveira Nogueira, Renata Giorgi, Marlos Cortez Sampaio, Robson L Melo, Catarina Teixeira, Daniel C PimentaAbstract:Abstract Echinometra Lucunter is an abundant sea urchin found in Brazilian waters. Accidents caused by this animal are common and are characterized by the penetration of the spines in the skin, which raises an inflammatory reaction through mechanical trauma as well as by the presumable action of toxins. Additionally, there have been reports of inflammatory reaction after the consumption of raw sea urchin eggs. In this work, we have isolated a peptide from E. Lucunter coelomic fluid that could elicit inflammatory reactions, such as paw edema, leukocyte recruitment and diminishment of the pain threshold. This peptide was termed Echinometrin . Moreover, the peptide administration was able to produce in vivo degranulation of mouse mast cells, in a dose–response manner. The peptide was ‘de novo’ sequenced by mass spectrometry and its synthetic analog could reproduce all the observed effects. Sequence alignment indicates that this peptide is comprised in vitellogenin, an abundant nutrient protein present in the gametogenic cells of sea urchins, making it possible that echinometrin would be a cryptide with pro-inflammatory effects.
-
cathepsin b x is secreted by Echinometra Lucunter sea urchin spines a structure rich in granular cells and toxins
Journal of Venomous Animals and Toxins Including Tropical Diseases, 2013Co-Authors: Juliana Mozer Sciani, Marta M Antoniazzi, Adriana Da Costa Neves, Daniel C PimentaAbstract:Background: Echinometra Lucunter is a common American sea urchin responsible for the majority of the marine accidents in Brazil. Although not lethal, these accidents are reported to be extremely painful. Recently, our group described the presence of toxins in its spines that contribute to the pathological reactions. Additionally, we have observed that the E. Lucunter spines can regenerate when broken. In the present work we evaluated the enzymatic activities of sea urchin spine extracts in order to identify an enzyme that could contribute not only to the toxicity, but also participate in the spine growth and regeneration. Results: The spine aqueous extract was tested for peptidase activity, with synthetic substrates, in the presence and absence of inhibitors and activators. For proper enzyme classification, the FRET-substrate cleavage pattern, pH-dependency activity and Western-blot analyses were performed. The spine extract was able to cleave Z-R-MCA and Abz-GIVRAK(Dnp)-OH following pre-incubation with DTT, and was inhibited by E-64. Furthermore, the double-peaked pH curve (5 and 7) and the cleavage site proportion (4:6, R↓A:A↓K) indicate the presence of both mono and dicarboxypeptidase activities. Moreover, in Western-blot analysis, the spine extract was positive for anti-cathepsin B antibody. Conclusions: E. Lucunter spines extracts presented a cysteine peptidase activity that was identified as cathepsin B/X that would participate in the remodeling and growth processes of the spine, as well as in the inflammatory response to the accident.
Daniel C Pimenta - One of the best experts on this subject based on the ideXlab platform.
-
preliminary molecular characterization of a proinflammatory and nociceptive molecule from the Echinometra Lucunter spines extracts
Journal of Venomous Animals and Toxins Including Tropical Diseases, 2017Co-Authors: Juliana Mozer Sciani, Bianca Cestari Zychar, Luis Roberto De Camargo Goncalves, Thiago De Oliveira Nogueira, Renata Giorgi, Daniel C PimentaAbstract:Sea urchins are animals commonly found on the Brazilian shoreline, being Echinometra Lucunter the most abundant species. Accidents caused by E. Lucunter have been reported as one of the most frequent in Brazil, and are characterized by intense pain and inflammation, consequence of spine puncture in the skin. In order to characterize such toxic effects, we isolated one molecule that caused inflammatory and nociceptive effects. E. Lucunter specimens were collected without gender distinction. Spines were removed and molecules were extracted, fractionated by RP-HPLC and assayed for inflammatory and nociceptive activity, in a biological-driven fractionation way, until the obtainment of one active molecule and its subsequent analysis by mass spectrometry (MS and MS/MS). For inflammation, intravital microscopy was performed on the mouse cremaster muscle, in order to evaluate rolled, adherent and migrating leukocytes. Paw edema was also evaluated. For the nociceptive activity, the paw pressure test was performed in rats. One molecule could be isolated and related to the inflammatory and nociceptive activity. Regarding inflammation, increase in adherent and migrating cells was observed in the cremaster muscle after the administration of the molecule. Corroborating the inflammatory response, paw edema was also observed, although only in 20% of controls and 20 min after injection. Additionally, this molecule was able to decrease significantly the pain threshold, characterizing hyperalgesia. This molecule was analyzed by mass spectrometry, and according to the exact molecular mass, isotopic distribution and fragmentation profile, it was possible to propose the molecular formula C29H48N3O10. One isolated molecule from the spine extract of E. Lucunter is able to elicit inflammation and hypernociception in animal models, which is in agreement with the effects observed in sea urchin accidents.
-
initial peptidomic profiling of brazilian sea urchins arbacia lixula lytechinus variegatus and Echinometra Lucunter
Journal of Venomous Animals and Toxins Including Tropical Diseases, 2016Co-Authors: Juliana Mozer Sciani, Jose Roberto Machado Cunha Da Silva, Andrews Krupinski Emerenciano, Daniel C PimentaAbstract:Background Sea urchins can be found throughout the Brazilian coast and are reported to be one of the major causes of marine accidents on the shoreline. Although not lethal, these accidents are reported to be extremely painful. In order to understand the toxinology of the Brazilian urchins, a peptidomic approach was performed aiming to characterize the naturally occurring peptides in both the coelomic fluid and the spine.
-
echinometrin a novel mast cell degranulating peptide from the coelomic liquid of Echinometra Lucunter sea urchin
Peptides, 2014Co-Authors: Juliana Mozer Sciani, Bianca Cestari Zychar, Luis Roberto De Camargo Goncalves, Thiago De Oliveira Nogueira, Renata Giorgi, Marlos Cortez Sampaio, Robson L Melo, Catarina Teixeira, Daniel C PimentaAbstract:Abstract Echinometra Lucunter is an abundant sea urchin found in Brazilian waters. Accidents caused by this animal are common and are characterized by the penetration of the spines in the skin, which raises an inflammatory reaction through mechanical trauma as well as by the presumable action of toxins. Additionally, there have been reports of inflammatory reaction after the consumption of raw sea urchin eggs. In this work, we have isolated a peptide from E. Lucunter coelomic fluid that could elicit inflammatory reactions, such as paw edema, leukocyte recruitment and diminishment of the pain threshold. This peptide was termed Echinometrin . Moreover, the peptide administration was able to produce in vivo degranulation of mouse mast cells, in a dose–response manner. The peptide was ‘de novo’ sequenced by mass spectrometry and its synthetic analog could reproduce all the observed effects. Sequence alignment indicates that this peptide is comprised in vitellogenin, an abundant nutrient protein present in the gametogenic cells of sea urchins, making it possible that echinometrin would be a cryptide with pro-inflammatory effects.
-
cathepsin b x is secreted by Echinometra Lucunter sea urchin spines a structure rich in granular cells and toxins
Journal of Venomous Animals and Toxins Including Tropical Diseases, 2013Co-Authors: Juliana Mozer Sciani, Marta M Antoniazzi, Adriana Da Costa Neves, Daniel C PimentaAbstract:Background: Echinometra Lucunter is a common American sea urchin responsible for the majority of the marine accidents in Brazil. Although not lethal, these accidents are reported to be extremely painful. Recently, our group described the presence of toxins in its spines that contribute to the pathological reactions. Additionally, we have observed that the E. Lucunter spines can regenerate when broken. In the present work we evaluated the enzymatic activities of sea urchin spine extracts in order to identify an enzyme that could contribute not only to the toxicity, but also participate in the spine growth and regeneration. Results: The spine aqueous extract was tested for peptidase activity, with synthetic substrates, in the presence and absence of inhibitors and activators. For proper enzyme classification, the FRET-substrate cleavage pattern, pH-dependency activity and Western-blot analyses were performed. The spine extract was able to cleave Z-R-MCA and Abz-GIVRAK(Dnp)-OH following pre-incubation with DTT, and was inhibited by E-64. Furthermore, the double-peaked pH curve (5 and 7) and the cleavage site proportion (4:6, R↓A:A↓K) indicate the presence of both mono and dicarboxypeptidase activities. Moreover, in Western-blot analysis, the spine extract was positive for anti-cathepsin B antibody. Conclusions: E. Lucunter spines extracts presented a cysteine peptidase activity that was identified as cathepsin B/X that would participate in the remodeling and growth processes of the spine, as well as in the inflammatory response to the accident.
-
pro inflammatory effects of the aqueous extract of Echinometra Lucunter sea urchin spines
Experimental Biology and Medicine, 2011Co-Authors: Juliana Mozer Sciani, Bianca Cestari Zychar, Luis Roberto De Camargo Goncalves, Thiago De Oliveira Nogueira, Renata Giorgi, Daniel C PimentaAbstract:The sea urchin, Echinometra Lucunter, can be found along the Western Central Atlantic shores. In Brazil, it is responsible by circa 50% of the accidents caused by marine animals. The symptoms usually surpass trauma and may be pathologically varied and last differently, ranging from spontaneous healing in a few days, to painful consequences lasting for weeks. In this work, we have mimicked the sea urchin accident by administering an aqueous extract of the spine into mice and rats and evaluated the pathophysiological developments. Our data clearly indicate that the sea urchin accident is indeed a pro-inflammatory event, triggered by toxins present in the spine that can cause edema and alteration in the leukocyte–endothelial interaction. Moreover, the spine extract was shown to exhibit a hyperalgesic effect. The extract is rich in proteins, as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but also contains other molecules that can be analyzed by reversed phase high-performance liquid...
Jose Roberto Machado Cunha Da Silva - One of the best experts on this subject based on the ideXlab platform.
-
Ocean acidification affects parameters of immune response and extracellular pH in tropical sea urchins Lytechinus variegatus and Echinometra luccunter
Aquatic Toxicology, 2016Co-Authors: Débora Alvares Leite Figueiredo, Paola Cristina Branco, Joao Carlos Shimada Borges, Douglas Amaral Dos Santos, Andrews Krupinski Emerenciano, Renata Stecca Iunes, Jose Roberto Machado Cunha Da SilvaAbstract:Abstract The rising concentration of atmospheric CO2 by anthropogenic activities is changing the chemistry of the oceans, resulting in a decreased pH. Several studies have shown that the decrease in pH can affect calcification rates and reproduction of marine invertebrates, but little attention has been drawn to their immune response. Thus this study evaluated in two adult tropical sea urchin species, Lytechinus variegatus and Echinometra Lucunter, the effects of ocean acidification over a period of 24 h and 5days, on parameters of the immune response, the extracellular acid base balance, and the ability to recover these parameters. For this reason, the phagocytic capacity (PC), the phagocytic index (PI), the capacity of cell adhesion, cell spreading, cell spreading area of phagocytic amebocytes in vitro, and the coelomic fluid pH were analyzed in animals exposed to a pH of 8.0 (control group), 7.6 and 7.3. Experimental pH’s were predicted by IPCC for the future of the two species. Furthermore, a recovery test was conducted to verify whether animals have the ability to restore these physiological parameters after being re-exposed to control conditions. Both species presented a significant decrease in PC, in the pH of coelomic fluid and in the cell spreading area. Besides that, Echinometra Lucunter showed a significant decrease in cell spreading and significant differences in coelomocyte proportions. The recovery test showed that the PC of both species increased, also being below the control values. Even so, they were still significantly higher than those exposed to acidified seawater, indicating that with the re-establishment of the pH value the phagocytic capacity of cells tends to restore control conditions. These results demonstrate that the immune system and the coelomic fluid pH of these animals can be affected by ocean acidification. However, the effects of a short-term exposure can be reversible if the natural values are re-established. Thus, the effects of ocean acidification could lead to consequences for pathogen resistance and survival of these sea urchin species.
-
initial peptidomic profiling of brazilian sea urchins arbacia lixula lytechinus variegatus and Echinometra Lucunter
Journal of Venomous Animals and Toxins Including Tropical Diseases, 2016Co-Authors: Juliana Mozer Sciani, Jose Roberto Machado Cunha Da Silva, Andrews Krupinski Emerenciano, Daniel C PimentaAbstract:Background Sea urchins can be found throughout the Brazilian coast and are reported to be one of the major causes of marine accidents on the shoreline. Although not lethal, these accidents are reported to be extremely painful. In order to understand the toxinology of the Brazilian urchins, a peptidomic approach was performed aiming to characterize the naturally occurring peptides in both the coelomic fluid and the spine.
-
the impact of rising sea temperature on innate immune parameters in the tropical subtidal sea urchin lytechinus variegatus and the intertidal sea urchin Echinometra Lucunter
Marine Environmental Research, 2013Co-Authors: Paola Cristina Branco, Joao Carlos Shimada Borges, Marinilce Fagundes Dos Santos, Bernard Ernesto Jensch, Jose Roberto Machado Cunha Da SilvaAbstract:Abstract Ocean temperatures are rising throughout the world, making it necessary to evaluate the impact of these temperature changes on sea urchins, which are well-known bioindicators. This study evaluated the effect of an increase in temperature on the immune response of the subtidal Lytechinus variegatus and the intertidal Echinometra Lucunter sea urchins. Both species were exposed to 20 (control), 25 and 30 °C temperatures for 24 h, 2, 7 and 14 days. Counting of coelomocytes and assays on the phagocytic response, adhesion and spreading of coelomocytes were performed. Red and colorless sphere cells were considered biomarkers for heat stress. Moreover, a significant decrease in the phagocytic indices and a decrease in both cell adhesion and cell spreading were observed at 25 and 30 °C for L. variegatus. For E. Lucunter, the only alteration observed was for the cell proportions. This report shows how different species of sea urchins respond immunologically to rising temperatures.
-
innate immune response in the sea urchin Echinometra Lucunter echinodermata
Journal of Invertebrate Pathology, 2008Co-Authors: Marcos Tucunduva De Faria, Jose Roberto Machado Cunha Da SilvaAbstract:Echinometra Lucunter, (Pinda) is a sea urchin encountered in the Brazilian coast and exposed to high and low temperatures related to low and high tides. Despite their great distribution and importance, few studies have been done on the biological function of their coelomocytes. Thus, Echinometra Lucunter perivisceral coelomocytes were characterized under optical and transmission electron microscopy. Phagocytic amoebocytes in the perivisceral coelom were labelled by injecting ferritin, and ferritin labelled phagocytic amoebocytes were found in the peristomial connective tissue after injecting India ink into the tissue, indicating the amoebocytes ability to respond to an inflammatory stimulus. Results showed that the phagocytic amoebocytes were the main inflammatory cells found in the innate immune response of E. Lucunter. While other works have recorded these phenomena in sea urchins found in moderate and constant temperature, this study reports on these same phenomena in a tropical sea urchin under great variation of temperature, thus providing new data to inflammatory studies in invertebrate pathology.
Luis Fernando Marquessantos - One of the best experts on this subject based on the ideXlab platform.
-
abcb1 and abcc1 like transporters in immune system cells from sea urchins Echinometra Lucunter and echinus esculentus and oysters crassostrea gasar and crassostrea gigas
Fish & Shellfish Immunology, 2017Co-Authors: Luis Fernando Marquessantos, Helene Hegaret, Leonardo Limasantos, Fernando Ramos Queiroga, Patricia Mirella Da SilvaAbstract:Abstract ABC transporters activity and expression have been associated with the multixenobiotic resistance phenotype (MXR). The activity of these proteins leads to a reduction in the intracellular concentration of several xenobiotics, thus reducing their toxicity. However, little attention has been given to the expression of ABC transporters in marine invertebrates and few studies have investigated their role in immune system cells of sea urchins and shellfish bivalves. The aim of the present study was to investigate the activity of the ABC transporters ABCB1 and ABCC1 in immune system cells of sea urchins (coelomocytes) and oysters (hemocytes) from different climatic regions (Brazil and France). Sea urchins and oysters were collected at Paraiba coast; Brazil ( Echinometra Lucunter and Crassostrea gasar ) and Rade of Brest; France ( Echinus esculentus and Crassostrea gigas ). Coelomocytes and hemocytes were stained with the ABC transporter substrate calcein-AM and dye accumulation analyzed under flow cytometry. Reversin 205 (ABCB1 transporter blocker) and MK571 (ABCC1 transporter blocker) were used as pharmacological tools to investigate ABC transporter activity. A different pattern of calcein accumulation was observed in coelomocytes: phagocytes > colorless spherulocytes > vibrate cells > red spherulocytes. The treatment with MK571 increased calcein fluorescence levels in coelomocytes from both species. However, reversin 205 treatment was not able to increase calcein fluorescence in E. esculentus coelomocytes. These data suggest that ABCC1-like transporter activity is present in both sea urchin species, but ABCB1-like transporter activity might only be present in E. Lucunter coelomocytes. The activity of ABCC1-like transporter was observed in all cell types from both bivalve species. However, reversin 205 only increased calcein accumulation in hyalinocytes of the oyster C. gasar , suggesting the absence of ABCB1-like transporter activity in all other cell types, including hyalinocytes from the oyster C. gigas. Additionally, our results showed that C. gigas exhibited higher activity of ABCC1-like transporter in all hemocyte types than C. gasar . The present work is the first to characterize ABCB1 and ABCC1-like transporter activity in the immune system cells of sea urchins E. Lucunter and E. esculentus and oysters. Our findings encourage the performing studies regarding ABC transporters activity/expression in immune system cells form marine invertebrates under stress conditions and the possible use of ABC transporters as biomarkers.
-
effects of salinity on the immune system cells of the tropical sea urchin Echinometra Lucunter
Journal of Experimental Marine Biology and Ecology, 2017Co-Authors: Thais Bezerra Mangeon Honorato, Raianna Boni, Patricia Mirella Da Silva, Luis Fernando MarquessantosAbstract:Abstract Human activities have caused climate changes and altered the salinity of the oceans. The reduction of the salinity is one of the factors that may limit the distribution and the survival of marine organisms. Coelomocytes are the immune system cells of the echinoderms and have been studied as biomarkers of stress. The aim of the present study was to investigate the effect of the salinity on the immune system cells of the tropical sea urchin Echinometra Lucunter . Animals were collected in Joao Pessoa coast (Brazilian Northeast). Animals or coelomocytes were exposed to different salinities (25, 35, 45) from 4 to 24 h. Phagocytic parameters, production of reactive oxygen species (ROS), mitochondrial activity and ABC transporter activity were analyzed. The phagocytic parameters did not change when animals or cells were exposed to a salinity of 25 or 45 in any time intervals monitored. However, the coelomocytes concentration increased when animals were exposed to the lower salinity. The levels of ROS were higher when cells were incubated at a salinity of 25 but lower when cells were kept at a salinity of 45. It was observed the loss of the mitochondrial inner membrane potential when coelomocytes were incubated at a salinity of 45. The activity of ABC transporters decreased when cells were incubated at the lowest salinity and increased when cells were incubated at the highest salinity tested. The present work shows that the immune system of the tropical sea urchins Ei Lucunter tolerates salinity changes from 25 to 45, and suggests two cellular parameters (ROS levels and ABC transporters activity) as potential biomarkers for the monitoring of the impact of environmental salinity changes.
-
involvement of abcb1 and abcc1 transporters in sea urchin Echinometra Lucunter fertilization
Molecular Reproduction and Development, 2012Co-Authors: Helena Lima Silvaneta, Elis Torrezan, Jocelmo Cassio De Araujo Leite, Tatiane Santigadelha, Luis Fernando MarquessantosAbstract:Fertilization is an ordered sequence of cellular interactions that promotes gamete fusion to form a new individual. Since the pioneering work of Oskar Hertwig conducted on sea urchins, echinoderms have contributed to the understanding of cellular and molecular aspects of the fertilization processes. Studies on sea urchin spermatozoa reported the involvement of a plasma membrane protein that belongs to the ABC proteins superfamily in the acrosome reaction. ABC transporters are expressed in membranes of eukaryotic and prokaryotic cells, and are associated with the transport of several compounds or ions across biomembranes. We aimed to investigate ABCB1 and ABCC1 transporter activity in sea urchin spermatozoa and their involvement in fertilization. Our results indicate that Echinometra Lucunter spermatozoa exhibit a low intracellular calcein accumulation (18.5% stained cells); however, the ABC blockers reversin205, verapamil, and MK571 increased dye accumulation (93.0–96.6% stained cells). We also demonstrated that pharmacologically blocking ABCB1 and ABCC1 decreased spermatozoa fertilizing capacity (70% inhibition), and this phenotype was independent of extracellular calcium. These data suggest that functional spermatozoa ABCB1 and ABCC1 transporters are crucial for a successful fertilization. Additional studies must be performed to investigate the involvement of membrane lipid homeostasis in the fertilization process. Mol. Reprod. Dev. 79: 861–869, 2012. © 2012 Wiley Periodicals, Inc.
-
extracellular ca2 influx is crucial for the early embryonic development of the sea urchin Echinometra Lucunter
Journal of Experimental Zoology, 2011Co-Authors: Jocelmo Cassio De Araujo Leite, Luis Fernando MarquessantosAbstract:The involvement of Ca2+ in the activation of eggs and in the first steps of the embryonic development of several species is a well-known phenomenon. An association between Ca2+ sources with the fate of the blastopore during embryonic development has been investigated by several authors. Ca2+ influx mediated by voltage-gated channels and Ca2+ mobilization from intracellular stores are the major sources of Ca2+ to egg activation and succeeding cell divisions. Studies on sea urchins embryonic development show that intracellular Ca2+ stores are responsible for egg activation and early embryogenesis. In the present work we investigated the involvement of extracellular Ca2+ in the first stages of the embryonic development of the sea urchin Echinometra Lucunter. Divalent cation chelators EDTA and EGTA strongly blocked the early embryonic development. Adding to this, we demonstrated the involvement of voltage-gated Ca2+ channels in E. Lucunter embryogenesis since Ca2+ channel blockers powerfully inhibited the early embryonic development. Our data also revealed that Ca2+ influx is crucial for embryonic development during only the first 40 min postfertilization. However, intracellular Ca2+ remains mandatory to embryonic development 40 min postfertilization, seen that both the intracellular Ca2+ chelator BAPTA-AM and calmodulin antagonists trifluoperazine and chlorpromazine inhibited the first stages of development when added to embryos culture 50 min postfertilization. Our work highlights the crucial role of extracellular Ca2+ influx through voltage-gated Ca2+ channels for the early embryonic development of the sea urchin E. Lucunter and characterizes an exception in the phylum Echinodermata. J. Exp. Zool. (Mol. Dev. Evol.) 318:123–133, 2012. © 2011 Wiley Periodicals, Inc.
-
characterization of functional activity of abcb1 and abcc1 proteins in eggs and embryonic cells of the sea urchin Echinometra Lucunter
Bioscience Reports, 2010Co-Authors: Monica Queiroz Vasconcelos De Souza, Elis Torrezan, Taissa Valeria Barros, Airlla Laana De Medeiros Cavalcanti, Regina Celia Bressan Queiroz De Figueiredo, Luis Fernando MarquessantosAbstract:ABC transporter (ATP-binding-cassette transporter) proteins have been strongly associated with the phenomenon of multidrug resistance in cancer cells. Furthermore, their physiological expression has been studied in many organisms, including bacteria, fungi, plants and vertebrate or invertebrate animals. Their widespread expression through the evolution demonstrates their relevance to the survival of living things. In the present study, we characterized the functional activity of ABCB1 and ABCC1 proteins in gametes and embryonic cells of the sea urchin Echinometra Lucunter . The ABC transporter proteins9 functional activity was up-regulated post-fertilization. Eggs and spermatozoa of E. Lucunter accumulated more C-AM (calcein acetoxymethyl ester), a fluorescent substrate of ABCB1 and ABCC1 proteins, than embryonic cells. Verapamil, reversin 205 and indomethacin were able to increase C-AM influx in eggs and embryos. However, verapamil and reversin 205 were more efficient than indomethacin, suggesting a predominance of ABCB1 protein over ABCC1 protein activity. Multidrug resistance modulating agents, at the concentration range that inhibited ABC transporter proteins, did not block the embryonic development until blastula stage. However, inhibition of ABCB1-mediated efflux by reversin 205 circumvented resistance of embryos to the antimitotic vinca alkaloid vinblastine. Embryonic development was more efficiently blocked when colchicine was previously added to eggs than to embryos 5 min after fertilization. This set of results suggests that these proteins act as a fundamental biochemical barrier conferring a protective physiological role against toxic xenobiotics in E. Lucunter embryos.
Alexandre Holanda Sampaio - One of the best experts on this subject based on the ideXlab platform.
-
sequenciamento parcial de uma lectina do ourico negro Echinometra Lucunter
Encontros Universitários da UFC, 2017Co-Authors: Tulio Jose Da Silva Marques, Romulo Farias Carneiro, Celso Shiniti Nagano, Alexandre Holanda SampaioAbstract:Lectinas sao proteinas de origem nao imune que reconhecem e se ligam a carboidratos de maneira especifica e reversivel. As lectinas sao moleculas de alto potencial biotecnologico com aplicacoes nas mais diversas areas e presentes em todos os organismos. Em equinodermos, poucas lectinas foram isoladas e caracterizadas a nivel estrutural e biologico. O ourico negro Echinometra Lucunter e uma especie amplamente distribuida em regioes tropicais. A lectina (Echinometra Lucunter agglutinin - ELA), alvo do presente estudo, foi purificada do fluido celomatico do ourico negro com o intuito de se determinar sua sequencia primaria de aminoacidos via espectrometria de massas. A especie foi coletada na Praia do Pacheco e acondicionada no Laboratorio de Biotecnologia Marinha-BIOMAR, o fluido celomatico dos individuos foi extraido e a lectina purificada por cromatografia liquida de afinidade em uma matriz de goma de xantana. A massa da proteina foi estimada por MALDI TOF e sua estrutura primaria foi obtida por ESI-MS/MS a partir da analise de peptideos tripticos. ELA exibiu massas medias entre 25 e 27 kDa com dois ions majoritarios de massas 25377 ±5 Da e 27291 ±5 Da. A partir da digestao triptica, obteve-se onze peptideos correspondendo a 54,5% de sua sequencia completa. Os dados indicaram grau de identidade em 47% e grau de similaridade em 50% para com uma proteina putativa da especie de ourico Strongylocentrotus purpuratus, alem de semelhancas com proteinas pertencentes ao grupo das interlectinas. Portanto, a lectina purificada do fluido celomatico de E. Lucunter assemelha-se a uma proteina presente em S. purpuratus. Estudos posteriores serao carreados de forma a concluir sua sequencia de aminoacidos, averiguar a definicao de ambas as proteinas como sendo interlectinas e suas aplicacoes biotecnologicas.
-
sequencia parcial de aminoacidos de uma lectina ligante de l ramnose a partir dos ovos de ourico negro Echinometra Lucunter
Encontros Universitários da UFC, 2017Co-Authors: Lucas De Vasconcelos Teixeira, Romulo Farias Carneiro, Alexandre Holanda Sampaio, Celso Shiniti NaganoAbstract:Lectinas sao proteinas de origem nao imune que se ligam de maneira reversivel a carboidratos especificos modificar quimicamente os mesmos. As lectinas RBLs (Lectina ligante de ramnose) pertencem a um grupo de lectinas que interagem especificamente a L- ramnose e alfa-galactose, em vez de beta-galactosideos, e nao necessitam de cations bivalentes ou grupamento tiol para exercer sua atividade hemaglutinante. O objetivo deste trabalho foi isolar, determinar a estrutura primaria e modificacoes pos traducionais, caracterizar estruturalmente uma lectina presente nos ovos de ourico negro Echinometra Lucunter. A lectina (ELEL) foi purificada atraves de cromatografia de afinidade em uma coluna de lactosil-agarose. A ELEL apresentou duas isoformas que foram separadas por cromatografia de fase reversa. Posteriormente uma analise por espectrometria de massa foi capaz de demonstrar uma pequena diferenca entre as massas moleculares de ELEL-1 e ELEL -2. Apos realizada a analise de peptideos foi possivel obter 12 peptideos, sendo 10 oriundos de digestao por tripsina e 2 por digestao com quimiotripsina. Quando comparados com a identidade da proteina SUEL o peptideo T2 apresentou 91,66% e 86,33% de similaridade e identidade, enquanto o peptideo Q1 apresenta 85% de identidade. O peptideo T3 representa o N-Terminal da ELEL que se apresenta bloqueado impossibilitando a analise por degradacao de Edman.
-
l rhamnose binding lectin from eggs of the Echinometra Lucunter amino acid sequence and molecular modeling
International Journal of Biological Macromolecules, 2015Co-Authors: Romulo Farias Carneiro, Celso Shiniti Nagano, Claudener S Teixeira, Arthur Alves De Melo, Alexandra Sampaio De Almeida, Benildo Sousa Cavada, Oscarina Viana De Sousa, B A M Rocha, Alexandre Holanda SampaioAbstract:Abstract An l -rhamnose-binding lectin named ELEL was isolated from eggs of the rock boring sea urchin Echinometra Lucunter by affinity chromatography on lactosyl-agarose. ELEL is a homodimer linked by a disulfide bond with subunits of 11 kDa each. The new lectin was inhibited by saccharides possessing the same configuration of hydroxyl groups at C-2 and C-4, such as l -rhamnose, melibiose, galactose and lactose. The amino acid sequence of ELEL was determined by tandem mass spectrometry. The ELEL subunit has 103 amino acids, including nine cysteine residues involved in four conserved intrachain disulfide bonds and one interchain disulfide bond. The full sequence of ELEL presents conserved motifs commonly found in rhamnose-binding lectins, including YGR, DPC and KYL. A three-dimensional model of ELEL was created, and molecular docking revealed favorable binding energies for interactions between ELEL and rhamnose, melibiose and Gb 3 (Galα1-4Galβ1-4Glcβ1-Cer). Furthermore, ELEL was able to agglutinate Gram-positive bacterial cells, suggesting its ability to recognize pathogens.
