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1.
Artículo en Inglés | MEDLINE | ID: mdl-36306998

RESUMEN

The enteric morphology, enteric microbiota structure and serum metabolomics of M. salmoides before and after infected by A. hydrophila were analysed to explore the pathogenic mechanism of A. hydrophila infection in M. salmoides. The results revealed that, after the infection of A. hydrophila, the villus boundary of largemouth bass became less obvious; the relative abundance of Proteobacteria and decreasing relative abundance of Tenericutes were increasing; genera relative abundance of putatively beneficial bacteria (Mycoplasma) were decreasing, whereas the genus Aeromonas increased after infection; serum metabolomic analysis showed that infection with A. hydrophila caused disorder to the metabolic processes of largemouth bass, particularly amino acid metabolism, and caused inflammation; several potential pathogen infection-related and significantly differential intestinal microbiota-related metabolite markers were identified, such as 6-hydroxy-5-methoxyindole glucuronide, zalcitabine, bilirubin, aciclovir. This study may provide new insights into the potential association between enteric microbiota and serum metabolism and the pathogenic mechanism of M. salmoides infected by A. hydrophila, providing a scientific basis for disease control in largemouth bass breeding.


Asunto(s)
Aeromonas , Lubina , Microbioma Gastrointestinal , Animales , Aeromonas hydrophila
2.
Fish Shellfish Immunol ; 103: 385-402, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32387478

RESUMEN

In the present study, we characterized tumor necrosis factor receptor-associated factor 2/7 (lcTRAF2/7) and TNFR1-associated death domain protein (lcTRADD) in Larimichthys crocea (L. crocea) and examined their expression profiles in tissues of Vibrio-challenged and unchallenged fish. The coding sequences of lcTRAF2, lcTRAF7, and lcTRADD were 1488, 2454, and 744 nucleotides, and they encoded proteins of 495, 344, and 248 amino acids, respectively. The results of phylogenetic analysis revealed that lcTRAF2, lcTRAF7, and lcTRADD were closest to Oplegnathus fasciatus (85%), Xiphophorus maculatus (97%), and Acanthochromis polyacanthus (65%), respectively. Multiple sequence alignment showed that lcTRAF2 and lcTRAF7 were highly conserved with other vertebrate TRAFs in their functional domains; however, lcTRADD was poorly conserved. The results of quantitative real-time polymerase chain reaction analysis indicated that lcTRAF2, lcTRAF7, and lcTRADD were constitutively expressed in the spleen, liver, kidney, heart, brain, gill, bladder, skin, fin, eye, and muscle. After challenging fish with Vibrio parahaemolyticus, the mRNA expression levels of lcTRAF2, lcTRAF7, and lcTRADD were upregulated in liver, spleen, and kidney. Immunofluorescence staining revealed that lcTRAF2 and lcTRADD were cytoplasmic in localization, whereas lcTRAF7 targeted both the cytoplasm and nucleus. In addition, the NF-κB protein level was upregulated after lipopolysaccharide stimulation in lcTRAF2, lcTRAF7, or lcTRADD overexpressing cells. Taken collectively, these results have improved our understanding of the functions of TRAF2, TRAF7, and TRADD in pathogenic infections in teleosts.


Asunto(s)
Enfermedades de los Peces/inmunología , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Perciformes/genética , Perciformes/inmunología , Secuencia de Aminoácidos , Animales , Proteínas de Peces/química , Perfilación de la Expresión Génica/veterinaria , Filogenia , Alineación de Secuencia/veterinaria , Proteína de Dominio de Muerte Asociada a Receptor de TNF/química , Proteína de Dominio de Muerte Asociada a Receptor de TNF/genética , Proteína de Dominio de Muerte Asociada a Receptor de TNF/inmunología , Péptidos y Proteínas Asociados a Receptores de Factores de Necrosis Tumoral/química , Péptidos y Proteínas Asociados a Receptores de Factores de Necrosis Tumoral/genética , Péptidos y Proteínas Asociados a Receptores de Factores de Necrosis Tumoral/inmunología , Vibriosis/inmunología , Vibriosis/veterinaria , Vibrio parahaemolyticus/fisiología
3.
Fish Shellfish Immunol ; 78: 259-269, 2018 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-29702237

RESUMEN

Glutathione peroxidases family is a key role in the antioxidant system in oxybiotic organisms for cell redox homeostasis. One of their members, phospholipid hydroperoxide glutathione peroxidase (GPx4) have unique monomeric structure and can directly react with complex lipid and membrane-bound peroxides under the presence of glutathione(GSH). In this paper, two complete GPx4 cDNAs (designated as LcGPx4a and LcGPx4b) from Larimichthys crocea are identified by rapid amplification of cDNA ends. The cDNA of LcGPx4a was consisted of a 5'-untranslated region (UTR) of 258 bp, a 3'-UTR of 330 bp, and an open reading frame (ORF) of 561 bp encoding 186 amino acid (aa) polypeptides. And the full-length sequence of LcGPx4b was 1164 bp with a 5'-UTR of 34 bp, a 3'-UTR of 551 bp and an ORF of 576 bp encoding a polypeptide of 191 aa residues with a predicted signal peptide of 15 aa. The characteristic selenocysteine insertion (SECIS) sequence was detected in the 3'UTR of the two sequences with 78 bp in length. The conserved active site of selenocysteine (Sec) encoded by TGA was also identified and formed a tetrad functional structure with glutamine, tryptophan, and asparagine in LcGPx4a and LcGPx4b. Two signature site motifs ("LRILAFPSNQFGNQEPG" and "LRILGFPCNQFGGQEPG") were both conserved in the deduced amino acid of LcGPx4a and LcGPx4b. The genomic structure analysis revealed that the two sequences both had 7 exons and 6 introns, and the Sec opal codon and SECIS element were located at the third and seventh exons, respectively. LcGPx4a and LcGPx4b both have a wide distribution in 9 tissues with various relative expression levels and a highest expression pattern in the liver. Under Vibrio parahaemolyticus challenge, their relative expression levels were altered in the liver, spleen, kidney, and head kidney but with different magnitudes and response time. LcGPx4a and LcGPx4b showed a significantly up-regulated trend in the spleen during experimental period. Above results suggested that LcGPx4a and LcGPx4b were two conserved immune molecules and might play a role in the immune response of fish with a tissue-depemdent manners.


Asunto(s)
Enfermedades de los Peces/inmunología , Regulación de la Expresión Génica/inmunología , Glutatión Peroxidasa/genética , Glutatión Peroxidasa/inmunología , Inmunidad Innata/genética , Perciformes/genética , Perciformes/inmunología , Secuencia de Aminoácidos , Animales , Antioxidantes/metabolismo , Secuencia de Bases , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica/veterinaria , Glutatión Peroxidasa/química , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/inmunología , Fosfolípido Hidroperóxido Glutatión Peroxidasa , Filogenia , Reacción en Cadena de la Polimerasa/veterinaria , ARN Mensajero/genética , ARN Mensajero/metabolismo , Distribución Aleatoria , Alineación de Secuencia/veterinaria , Vibriosis/inmunología , Vibrio parahaemolyticus/fisiología
4.
Fish Shellfish Immunol ; 71: 411-422, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-28964863

RESUMEN

Glutathione peroxidases, a vital family of antioxidant enzymes in oxybiotic organisms, are involved in anti-pathogen immune response. In this study, two complete selenium-dependent glutathione peroxidase 1 cDNAs (designated as LcGPx1a and LcGPx1b) were obtained from the large yellow croaker Larimichthys crocea by rapid amplification of cDNA ends. The full-length sequence of LcGPx1a was 917 bp with a 5'-untranslated region (UTR) of 52 bp, a 3'-UTR of 289 bp, and an open reading frame of 576 bp encoding 191 amino acid (aa) polypeptides. The cDNA of LcGPx1b was composed of 884 bp with a 5'-UTR of 59 bp, a 3'-UTR of 258 bp, and an open reading frame of 567 bp encoding 188 aa polypeptides. The conserved selenocysteine insertion sequence was detected in the 3'-UTR of both isoforms, which can classify types I and II. Protein sequence analysis revealed that both isoforms included a selenocysteine encoded by an opal codon (TGA) and formed the functioning tetrad site with glutamine, tryptophan, and asparagine. Three conservative motifs, including one active site motif ("GKVVLIENVASLUGTT") and two signature site motifs ("LVILGVPCNQFGHQENC" and "V(A/S)WNFEKFLI"), were conserved both in sequence and location. Multiple alignments revealed that they exhibited a high level of identities with GPx1 from other organisms, especially in the abovementioned conserved amino acid sequence motifs. Tissue expression analysis indicated that LcGPx1a and LcGPx1b had a wide distribution in nine tissues with various abundances. The transcript level of LcGPx1a was not significantly different among the nine tissues, whereas that of LcGPx1b was higher in the kidney and head kidney than in the other tissues. After Vibrio parahaemolyticus stimulation, the expression levels of LcGPx1a and LcGPx1b were unanimously altered in the liver, spleen, kidney, and head kidney but with different magnitudes and response time. LcGPx1a and LcGPx1b showed distinct expression trends in the liver, where LcGPx1b was induced and LcGPx1a was depressed in response to pathogen infection. These results indicate that LcGPx1a and LcGPx1b display functional diversities and play crucial roles in mediating the immune response of fish.


Asunto(s)
Enfermedades de los Peces/inmunología , Regulación de la Expresión Génica/inmunología , Glutatión Peroxidasa/genética , Glutatión Peroxidasa/inmunología , Inmunidad Innata/genética , Perciformes/genética , Perciformes/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica , Glutatión Peroxidasa/química , Isoenzimas/química , Isoenzimas/genética , Isoenzimas/inmunología , Filogenia , Alineación de Secuencia/veterinaria , Vibriosis/inmunología , Vibrio parahaemolyticus/fisiología , Glutatión Peroxidasa GPX1
5.
Ecotoxicol Environ Saf ; 139: 18-26, 2017 May.
Artículo en Inglés | MEDLINE | ID: mdl-28092736

RESUMEN

The present study explored the possible preventive effects of blue light emitting diodes (LEDs) on cadmium (Cd)-induced oxidative stress and immunotoxicity in zebrafish. To this end, zebrafish were exposed to a white fluorescent bulb or blue LEDs (LDB, peak at 450nm, at an irradiance of 0.9W/m2), and 0 or 30µgL-1 waterborne Cd for 5 weeks. Growth performance, survival rate, and hepatic histology, ultrastructure, antioxidant and innate immune responses were determined in zebrafish. Cd exposure alone reduced growth and survival rate, and induced oxidative damage and changes in histology and ultrastructure. However, Cd exposure in combination with LDB apparently relieved these negative effects. The alleviation of adverse effects might result from the up-regulation of antioxidant and innate immune genes at transcriptional, translational, or post-translational levels. Cd exposure alone dramatically enhanced mRNA levels of nuclear transcription factor κB (NF-κB) and E2-related factor (Nrf2). However, compared to Cd exposure alone, Cd exposure in combination with LDB apparently down-regulated both genes. Taken together, our results suggest that chronic Cd exposure induced a negative effect on zebrafish, possibly involved in NF-κB-induced immunotoxicity and Nrf2-induced oxidative stress. Finally, for the first time, our data demonstrated that LDB could protect fish against Cd toxicity.


Asunto(s)
Antioxidantes , Cadmio/toxicidad , Inmunidad Innata , Luz , Hígado/efectos de los fármacos , Estrés Oxidativo , Pez Cebra/metabolismo , Animales , Antioxidantes/metabolismo , Regulación hacia Abajo , Exposición a Riesgos Ambientales , Inmunidad Innata/efectos de los fármacos , Inmunidad Innata/genética , Hígado/metabolismo , Hígado/patología , Hígado/ultraestructura , Factor 2 Relacionado con NF-E2/genética , Factor 2 Relacionado con NF-E2/metabolismo , FN-kappa B/genética , FN-kappa B/metabolismo , ARN Mensajero/metabolismo , Regulación hacia Arriba , Pez Cebra/genética , Pez Cebra/crecimiento & desarrollo , Proteínas de Pez Cebra/genética , Proteínas de Pez Cebra/metabolismo
6.
Aquat Toxicol ; 182: 113-119, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27888766

RESUMEN

Up to date, little information is available on effects of circadian rhythm on metal-induced toxicity in fish. In this study, zebrafish were acutely exposed to 0.97mgL-1 cadmium for 12h either at ZT0 (the light intensity began to reached maximum) or at ZT12 (light intensity began to reached minimum) to evaluate the temporal sensitivity of oxidative stress and inflammatory responses in the brain of zebrafish. Profiles of responses of some genes at mRNA, protein and activity levels were different between ZT0 and ZT12 in the normal water. Exposure to Cd induced contrary antioxidant responses and similar inflammatory responses between ZT0 and ZT12. However, the number of inflammatory genes which were up-regulated was significantly greater at ZT12 than at ZT0. And, the up-regulated inflammatory genes were more responsive at ZT12 than at ZT0. At ZT12, antioxidant genes were down-regulated at mRNA, protein and activity levels. Contrarily, antioxidant genes were not affected at mRNA levels but activated at the protein and/or activity levels at ZT0. Reactive oxygen species (ROS) sharply increased and remained relatively stable when fish were exposed to Cd at ZT12 and ZT0, respectively. Positive correlations between ROS levels and mRNA levels of nuclear transcription factor κB (NF-κB) and between mRNA levels of NF-κB and its target genes were observed, suggesting that ROS may play an essential role in regulating the magnitude of inflammatory responses. Taken together, oxidative stress and immunotoxicity in the brain were more serious when fish were exposed to Cd in the evening than in the morning, highlighting the importance of circadian rhythm in Cd-induced neurotoxicity in fish.


Asunto(s)
Antioxidantes/metabolismo , Encéfalo/efectos de los fármacos , Cadmio/toxicidad , Ritmo Circadiano/fisiología , Inflamación/inducido químicamente , Contaminantes Químicos del Agua/toxicidad , Pez Cebra/fisiología , Animales , Biomarcadores/metabolismo , Encéfalo/metabolismo , Regulación hacia Abajo/efectos de los fármacos , Inflamación/metabolismo , FN-kappa B/metabolismo , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Pruebas de Toxicidad Aguda , Regulación hacia Arriba/efectos de los fármacos
7.
J Exp Zool A Ecol Genet Physiol ; 325(7): 453-66, 2016 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-27455909

RESUMEN

Gonadotropin-releasing hormone (GnRH) plays a vital role in the regulation of reproduction through interaction with a specific receptor (the GnRH receptor). In this study, the GnRH receptor gene from the cuttlefish Sepiella japonica (SjGnRHR) was identified and characterized. The cloned full-length SjGnRHR cDNA was 1,468 bp long and contained a 1,029 bp open reading frame encoding 342 amino acid residues, 8 bp of 5' untranslated regions (UTR), and 431 bp of 3' UTR. The putative protein was predicted to have a molecular weight of 38.75 kDa and an isoelectric point of 9.47. In addition, this protein was identified as belonging to the rhodopsin-type (class A) G protein-coupled receptor family. The predicted amino acid sequence contained two N-linked glycosylation sites and 18 phosphorylation sites. Multiple sequence alignment, phylogenetic tree analysis, and three-dimensional structure modeling were conducted to clarify SjGnRHR bioinformatics characteristics. In vitro SjGnRHR expression was carried out using HEK293 cells and the pEGFP-N1 plasmid, to verify the transmembrane properties of this protein. The interaction between the S. japonica GnRH receptor and its ligand was clarified using internalization analysis. SjGnRHR transcriptional quantification confirmed the wide distribution of SjGnRHR in various S. japonica mature tissues. In addition, the transcriptional profile of SjGnRHR in the female brain and ovary during gonadal development was analyzed. Results indicate that GnRHR may be involved in diverse S. japonica physiological functions, especially in the control of reproduction.


Asunto(s)
Decapodiformes/metabolismo , Expresión Génica , Receptores LHRH/química , Secuencia de Aminoácidos , Animales , Clonación Molecular , Decapodiformes/fisiología , Femenino , Humanos , Masculino , Modelos Moleculares , Especificidad de Órganos , Filogenia , Receptores LHRH/genética , Receptores LHRH/metabolismo , Receptores LHRH/fisiología , Reproducción , Alineación de Secuencia
8.
Fish Physiol Biochem ; 42(6): 1595-1607, 2016 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-27380381

RESUMEN

AMP-activated protein kinase (AMPK) is a highly conserved and multi-functional protein kinase that plays important roles in both intracellular energy balance and cellular stress response. In the present study, molecular characterization, tissue distribution and gene expression levels of the AMPK α1 and α2 genes from turbot (Scophthalmus maximus) under salinity stress are described. The complete coding regions of the AMPK α1 and α2 genes were isolated from turbot through degenerate primers in combination with RACE using muscle cDNA. The complete coding regions of AMPK α1 (1722 bp) and α2 (1674 bp) encoded 573 and 557 amino acids peptides, respectively. Multiple alignments, structural analysis and phylogenetic tree construction indicated that S. maximus AMPK α1 and α2 shared a high amino acid identity with other species, especially fish. AMPK α1 and α2 genes could be detected in all tested tissues, indicating that they are constitutively expressed. Salinity challenges significantly altered the gene expression levels of AMPK α1 and α2 mRNA in a salinity- and time-dependent manners in S. maximus gill tissues, suggesting that AMPK α1 and α2 played important roles in mediating the salinity stress in S. maximus. The expression levels of AMPK α1 and α2 mRNA were a positive correlation with gill Na+, K+-ATPase activities. These findings will aid our understanding of the molecular mechanism of juvenile turbot in response to environmental salinity changes.


Asunto(s)
Proteínas Quinasas Activadas por AMP/genética , Proteínas de Peces/genética , Peces Planos/genética , Salinidad , Estrés Fisiológico/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , ADN Complementario/genética , Proteínas de Peces/metabolismo , Peces Planos/metabolismo , Expresión Génica , Branquias/enzimología , Filogenia , Isoformas de Proteínas/genética , ARN Mensajero/metabolismo , ATPasa Intercambiadora de Sodio-Potasio/metabolismo
9.
Ecotoxicol Environ Saf ; 132: 345-52, 2016 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-27362492

RESUMEN

The aim of the present study was to evaluate the effects of acute inorganic Hg exposure (0, 32 and 64µgHgL(-1)) on lipid peroxidation, activities and gene expression of antioxidant enzymes (Cu/Zn-SOD, CAT, GPx, GR and GST), and mRNA levels of the Keap1-Nrf2 signaling molecules at different exposure times (6h, 12h, 24h, 48h, and 96h) in the liver of large yellow croaker Pseudosciaena crocea. The results showed that lipid peroxidation was sharply reduced by 32µg Hg L(-1) during 6-12h before returning to control levels. Similarly, lipid peroxidation was significantly reduced during 6-12h followed by a sharp increase towards the end of the exposure in the 64µgHgL(-1) group. There was a negative relationship between lipid peroxidation and antioxidant enzyme activities, and positive relationship between activities and gene expression of antioxidant enzymes, suggesting that the changes at a molecular level may underlie enzymatic level and accordingly affect hepatic lipid peroxidation. Obtained results also showed a coordinated transcriptional regulation of antioxidant genes, suggesting that Nrf2 is required for the protracted induction of these genes. Furthermore, a negative relationship between the mRNA levels of Nrf2 and Keap1 indicated that Keap1 may play an important role in switching off the Nrf2 response. In conclusion, this is the first study to elucidate effects of waterborne Hg on antioxidant system in large yellow croaker through the Keap1-Nrf2 pathway, which will aid our understanding of the molecular mechanisms of waterborne heavy metal on antioxidant responses in fish.


Asunto(s)
Mercurio/toxicidad , Factor 2 Relacionado con NF-E2/fisiología , Estrés Oxidativo/efectos de los fármacos , Perciformes/metabolismo , Animales , Antioxidantes/metabolismo , Regulación de la Expresión Génica , Peroxidación de Lípido/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Mercurio/metabolismo , Compuestos de Mercurio/metabolismo , Compuestos de Mercurio/toxicidad , Factor 2 Relacionado con NF-E2/genética , Oxidación-Reducción/efectos de los fármacos , Distribución Aleatoria , Transducción de Señal/efectos de los fármacos
10.
Fish Shellfish Immunol ; 55: 535-49, 2016 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-27346150

RESUMEN

In this study, we sequenced and characterized melanoma differentiation-associated antigen 5 (LcMDA5), laboratory of genetics and physiology 2 (LcLGP2) and mitochondrial antiviral signaling protein (LcMAVS) from large yellow croaker (Larimichthys crocea). The LcMDA5 encodes 969 amino acids and contains two caspase-associated and recruitment domains (CARDs), a DExDc (DExD/H box-containing domain), a HELICc (helicase superfamily C-terminal domain) and a C-terminal regulatory domain (RD). The LcLGP2 encodes 679 amino acids and contains a DExDc, a HELICc and a RD. The LcMAVS encodes 512 amino acids and contains a CARD, a proline-rich domain, a transmembrane helix domain and a putative TRAF2-binding motif ((269)PVQDT(273)). Phylogenetic analyses showed that all the three genes of large yellow croaker are clustered together with their counterparts from other teleost fishes. The Real-time PCR analyses showed that all the three genes were found to be constitutively expressed in all examined tissues in large yellow croaker, but all with relatively low expression levels. Expression analyses showed that the three genes were all rapidly and significantly upregulated in vivo after poly (I:C) challenge in peripheral blood, liver, spleen and head kidney tissues. The results indicate that the LcMDA5, LcLGP2 and LcMAVS might play important roles in antiviral immune responses.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas de Peces/genética , Regulación de la Expresión Génica/inmunología , Inmunidad Innata , Helicasa Inducida por Interferón IFIH1/genética , Perciformes/genética , ARN Helicasas/genética , Proteínas Adaptadoras Transductoras de Señales/química , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Secuencia de Aminoácidos , Animales , Proteínas de Peces/química , Proteínas de Peces/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Helicasa Inducida por Interferón IFIH1/química , Helicasa Inducida por Interferón IFIH1/metabolismo , Perciformes/inmunología , Perciformes/metabolismo , Poli I-C/farmacología , Conformación Proteica , ARN Helicasas/química , ARN Helicasas/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Alineación de Secuencia
11.
Fish Shellfish Immunol ; 52: 1-8, 2016 May.
Artículo en Inglés | MEDLINE | ID: mdl-26940795

RESUMEN

We evaluated the effects of acute Zn exposure (4 and 8 mg L(-1) Zn) on lipid peroxidation, and activities and mRNA levels of antioxidant enzyme genes (Cu/Zn-SOD, CAT, GPx, and GR), and gene expression of the Nrf2-Keap1 signaling molecule at different exposure times (0, 6, 12, 24, 48, and 96 h) in the spleen of large yellow croaker. Lipid peroxidation remained relatively constant during 6-48 h and 6-24 h and sharply increased at 96 h and during 48-96 h in fish exposed to 4 and 8 mg L(-1) Zn, respectively. Activities of all tested enzymes increased during the early stage of exposure and decreased towards the end of the exposure in both groups. However, mRNA levels of antioxidant enzyme genes were dramatically up-regulated by 4 and 8 mg L(-1) Zn during the late stage of exposure. During the early stage of exposure for 6 h, the 8 mg L(-1) Zn exposure sharply increased mRNA levels of Cu/Zn-SOD, CAT, GPx1b, Nrf2, and Keap1, whereas, the 4 mg L(-1) Zn exposure did not significantly affect the expression of these genes. Our data also showed positive relationships between Nrf2 expression and mRNA levels of its target genes, suggesting that Nrf2 was required for the protracted induction of these genes. Furthermore, a sharp increase in Keap1 expression levels was observed in fish exposed to 4 mg L(-1) at 96 h, and 8 mg L(-1) at 6, 48, and 96 h. In conclusion, the present study demonstrated that Zn-induced antioxidant defenses were involved in modifications at enzymatic and transcriptional levels and the transcriptional regulation of the Nrf2-Keap1 signaling molecule; these results may contribute to the understanding of mechanisms that maintain the correct redox balance in the immune organ of the large yellow croaker.


Asunto(s)
Proteínas de Peces/genética , Regulación de la Expresión Génica/efectos de los fármacos , Proteína 1 Asociada A ECH Tipo Kelch/genética , Factor 2 Relacionado con NF-E2/genética , Perciformes/genética , Bazo/efectos de los fármacos , Zinc/toxicidad , Animales , Antioxidantes/metabolismo , Proteínas de Peces/metabolismo , Perfilación de la Expresión Génica , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Peroxidación de Lípido , Factor 2 Relacionado con NF-E2/metabolismo , Perciformes/inmunología , Perciformes/metabolismo , Bazo/metabolismo , Contaminantes Químicos del Agua/toxicidad
12.
Fish Shellfish Immunol ; 51: 1-8, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26892794

RESUMEN

Long noncoding RNAs (lncRNAs) are a class of transcripts that longer than 200 bp and do not encode proteins. Recent genome-wide studies of vertebrate transcriptomes have annotated lncRNAs that are expressed in various tissues and development stages. The draft genome and several transcriptome sequencing data sets have been collected for the study of protein-coding genes in large yellow croaker (Larimichthys crocea), but little is known about the expression and functional roles of lncRNAs in this species. In order to obtain a catalog of lncRNAs for large yellow croaker, several RNA-seq datasets were integrated from various tissues including egg, muscle, liver, and spleen. A total of 48,953 high-confidence transcripts were reconstructed in 38,017 loci, recovering the most of expressed reference transcripts while thousands of novel expressed loci have been identified. The tissue expression profile revealed that most lncRNAs were specifically enriched in different tissues. A stringent set of 210 lncRNAs were identified as being specifically expressed in spleen and potentially involved in immune response. Our study first systematically identify lncRNAs in large yellow croaker, benefiting the future genomic study of this species.


Asunto(s)
Inmunidad Innata , Perciformes/genética , ARN Largo no Codificante/genética , Transcriptoma , Animales , Perfilación de la Expresión Génica/veterinaria , Especificidad de Órganos , Perciformes/inmunología , Perciformes/metabolismo , ARN Largo no Codificante/metabolismo , Análisis de Secuencia de ARN/veterinaria
13.
Environ Sci Pollut Res Int ; 23(8): 7529-42, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26728289

RESUMEN

Large yellow croaker (LYC) cage farming is a rapidly developing industry in the coastal areas of the East China Sea. However, little is known about the environmental nutrient loadings resulting from the current aquaculture practices for this species. In this study, a nitrogenous waste model was developed for LYC based on thermal growth and bioenergetic theories. The growth model produced a good fit with the measured data of the growth trajectory of the fish. The total, dissolved and particulate nitrogen outputs were estimated to be 133, 51 and 82 kg N tonne(-1) of fish production, respectively, with daily dissolved and particulate nitrogen outputs varying from 69 to 104 and 106 to 181 mg N fish(-1), respectively, during the 2012 operational cycle. Greater than 80 % of the nitrogen input from feed was predicted to be lost to the environment, resulting in low nitrogen retention (<20 %) in the fish tissues. Ammonia contributed the greatest proportion (>85 %) of the dissolved nitrogen generated from cage farming. This nitrogen loading assessment model is the first to address nitrogenous output from LYC farming and could be a valuable tool to examine the effects of management and feeding practices on waste from cage farming. The application of this model could help improve the scientific understanding of offshore fish farming systems. Furthermore, the model predicts that a 63 % reduction in nitrogenous waste production could be achieved by switching from the use of trash fish for feed to the use of pelleted feed.


Asunto(s)
Explotaciones Pesqueras , Modelos Teóricos , Nitrógeno/análisis , Perciformes/crecimiento & desarrollo , Contaminantes Químicos del Agua/análisis , Amoníaco/análisis , Animales , China , Perciformes/metabolismo
14.
Aquat Toxicol ; 172: 21-9, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26765083

RESUMEN

The hypothesis tested in the present study was that Zn acclimation will alleviate high Zn induced oxidative stress in large yellow croaker Pseudosciaena crocea. To the end, fish were pre-exposed to 0 and 2mgZnL(-1) for 48h and then exposed to 0 and 10mgZnL(-1) for 48h. Lipid peroxidation, activities and mRNA levels of antioxidant enzyme genes (Cu/Zn-SOD, CAT, GPx and GR), and gene expressions of Nrf2-Keap1 signaling molecules at different exposure time (12h, 24h and 48h) were determined in the liver and spleen of large yellow croaker. 10mgZnL(-1) exposure alone enhanced lipid peroxidation in the liver during 12-48h and in the spleen during 24-48h. Although 2mgZnL(-1) pre-exposure did not affect lipid peroxidation, 2mgZnL(-1) pre-exposure mitigated high Zn induced oxidative stress. The positive effect of Zn acclimation could be attributed to the up-regulated expression and activities of antioxidant enzyme genes under high Zn stress. Obtained results also showed a coordinated transcriptional regulation of antioxidant genes, suggesting that Nrf2 is required for the protracted induction of these genes. Besides, the sharp increase in Keap1 expression levels would support its role in switching off Nrf2 response. In conclusion, Zn acclimation mitigated high Zn-induced oxidative stress in large yellow croker, emphasizing a central role of transcription factor Nrf2 in the process.


Asunto(s)
Regulación de la Expresión Génica/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Perciformes/fisiología , Zinc/metabolismo , Zinc/toxicidad , Aclimatación/genética , Animales , Proteínas de Peces/genética , Peroxidación de Lípido/efectos de los fármacos , Hígado/efectos de los fármacos , Oxidorreductasas/genética , Oxidorreductasas/metabolismo , Perciformes/metabolismo , Bazo/efectos de los fármacos , Contaminantes Químicos del Agua/toxicidad
15.
Gene ; 576(1 Pt 3): 458-65, 2016 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-26519998

RESUMEN

Gonadotropin-releasing hormone III (GnRH3) is considered to be a key neurohormone in fish reproduction control. In the present study, the cDNA and genomic sequences of GnRH3 were cloned and characterized from large yellow croaker Larimichthys crocea. The cDNA encoded a protein of 99 amino acids with four functional motifs. The full-length genome sequence was composed of 3797 nucleotides, including four exons and three introns. Higher identities of amino acid sequences and conserved exon-intron organizations were found between LcGnRH3 and other GnRH3 genes. In addition, some special features of the sequences were detected in partial species. For example, two specific residues (V and A) were found in the family Sciaenidae, and the unique 75-72 bp type of the open reading frame 2 and 3 existed in the family Cyprinidae. Analysis of the 2576 bp promoter fragment of LcGnRH3 showed a number of transcription factor binding sites, such as AP1, CREB, GATA-1, HSF, FOXA2, and FOXL1. Promoter functional analysis using an EGFP reporter fusion in zebrafish larvae presented positive signals in the brain, including the olfactory region, the terminal nerve ganglion, the telencephalon, and the hypothalamus. The expression pattern was generally consistent with the endogenous GnRH3 GFP-expressing transgenic zebrafish lines, but the details were different. These results indicate that the structure and function of LcGnRH3 are generally similar to the other teleost GnRH3 genes, but there exist some distinctions among them.


Asunto(s)
Hormona Liberadora de Gonadotropina/genética , Perciformes/genética , Regiones Promotoras Genéticas , Ácido Pirrolidona Carboxílico/análogos & derivados , Secuencia de Aminoácidos , Animales , Animales Modificados Genéticamente , Secuencia de Bases , Hormona Liberadora de Gonadotropina/química , Datos de Secuencia Molecular , Sistemas de Lectura Abierta , Ácido Pirrolidona Carboxílico/química , Homología de Secuencia de Aminoácido
16.
Mitochondrial DNA A DNA Mapp Seq Anal ; 27(5): 3122-3, 2016 09.
Artículo en Inglés | MEDLINE | ID: mdl-25648920

RESUMEN

In this study, we determined the complete mitochondrial genome of the little squid (Loliolus (Nipponololigo) uyii). The genome was 17,134 bp in length and contained 13 protein-coding genes,22 transfer RNA genes, 2 ribosomal RNA genes and 3 main non-coding regions. The composition and order of genes were similar to most other invertebrates. The overall base composition of L. uyii is A 39.98%, C 18.65 %, T 32.39% and G 8.98%, with a high A + T bias of 72.37%. All of the three control regions (CR) contain termination-associated sequences and conserved sequence blocks. These mitogenome sequence data would play an important role in the investigation of phylogenetic relationship, taxonomic resolution and phylogeography of the Loliginidae.


Asunto(s)
Decapodiformes/genética , Genoma Mitocondrial , Animales , Composición de Base , Decapodiformes/clasificación , Sistemas de Lectura Abierta , Filogenia , ARN Ribosómico/genética , ARN de Transferencia/genética , ARN no Traducido/genética
17.
Mitochondrial DNA A DNA Mapp Seq Anal ; 27(5): 3337-8, 2016 09.
Artículo en Inglés | MEDLINE | ID: mdl-25703845

RESUMEN

In this study, we determined the complete mitochondrial genome of the Loligo opalescence. The genome was 17,370 bp in length and contained 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and 3 main non-coding regions. The composition and order of genes, were similar to most other invertebrates. The overall base composition of L. opalescence is A 38.62%, C 19.40%, T 32.37% and G 9.61%, with a highly A + T bias of 70.99%. All of the three control regions (CR) contain termination-associated sequences and conserved sequence blocks. This mitogenome sequence data would play an important role in the investigation of phylogenetic relationship, taxonomic resolution and phylogeography of the Loliginidae.


Asunto(s)
Genoma Mitocondrial , Loligo/genética , Animales , Composición de Base , Codón , Secuencia Conservada , Orden Génico , Genes Mitocondriales , Tamaño del Genoma , Loligo/clasificación , Sistemas de Lectura Abierta , Secuencias Reguladoras de Ácidos Nucleicos , Análisis de Secuencia de ADN , Secuenciación Completa del Genoma
18.
Mitochondrial DNA A DNA Mapp Seq Anal ; 27(5): 3335-6, 2016 09.
Artículo en Inglés | MEDLINE | ID: mdl-25707415

RESUMEN

In this study, we determined the complete mitochondrial genome of the Illex argentinus. The genome was 20,278 bp in length and contained 18 protein-coding genes, 23 transfer RNA genes, 2 ribosomal RNA genes and 3 main non-coding regions. The composition and order of genes were different to some other invertebrates. The overall base composition of I. argentinus was A 39.23%, C 17.50%, T 33.71% and G 9.56%, with a highly A + T bias of 72.94%. All of the three control regions (CR) contained termination-associated sequences and conserved sequence blocks. This mitogenome sequence data would play an important role in the taxonomic resolution and phylogeography of the Ommastrephidae.


Asunto(s)
Decapodiformes/genética , Genoma Mitocondrial , Animales , Composición de Base , Codón , Decapodiformes/clasificación , Orden Génico , Genes Mitocondriales , Tamaño del Genoma , Sistemas de Lectura Abierta , Secuencias Reguladoras de Ácidos Nucleicos , Análisis de Secuencia de ADN , Secuenciación Completa del Genoma
19.
Mitochondrial DNA A DNA Mapp Seq Anal ; 27(4): 2723-4, 2016 07.
Artículo en Inglés | MEDLINE | ID: mdl-26104158

RESUMEN

In this study, we determined the complete mitochondrial genome of the little squid (Loligo duvaucelii). The genome is 17,413 bp in length, containing 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and 3 main non-coding regions. The overall base composition of L. duvaucelii is 40.01% A, 32.33% T, 19.14% C and 8.52% G, with a high A + T bias of 72.34%. All of the three control regions (CR) contain termination-associated sequences and conserved sequence blocks. Here, we describe a phylogenetic analysis of 10 species of Cephalopoda based on the complete mitochondrial genome, the result showed that the Loliolus uyii is most closely related to L. duvaucelii. This mitogenome sequence data would play an important role in the investigation of phylogenetic relationship, taxonomic resolution and phylogeography of the Cephalopoda.


Asunto(s)
Genoma Mitocondrial/genética , Loligo/genética , Animales , Composición de Base/genética , ADN Mitocondrial/genética , Orden Génico/genética , Loligo/clasificación , Filogenia , ARN Ribosómico/genética , ARN de Transferencia/genética , Análisis de Secuencia de ADN
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