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1.
Food Chem ; 403: 134309, 2023 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-36191413

RESUMO

To demonstrate the roles of bacteria in fish protein degradation and related quality changes, three major grass carp spoilage bacteria were individually inoculated into grass carp flesh, and their effects on protein molecules, muscle structures, and quality indices were evaluated through peptidomics, optical microscopy and transmission electron microscope, and texture and water distribution analyses, respectively. Results showed that Pseudomonas putida degraded obscurin, nebulin, and titin, caused disarrangement of myofilaments and fragmented myofibers, and induced great loss of free water in muscle. Shewanella putrefaciens was active in hydrolyzing collagen and degraded both thick and thin filament proteins. Mutual separation of myofibers and severe texture softening were also observed in S. putrefaciens-inoculated samples. Aeromonas rivipollensis degraded myosin heavy chain and some thin filament proteins but less affected muscle microstructure and quality indices. Therefore, this study revealed the mechanisms of bacteria-induced grass carp protein degradation and provided guidance for developing quality control strategies.


Assuntos
Carpas , Animais , Carpas/metabolismo , Proteínas de Peixes/metabolismo , Proteólise , Miofibrilas/metabolismo , Água/metabolismo
2.
J Sci Food Agric ; 103(1): 298-307, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-35861049

RESUMO

BACKGROUND: Apoptosis activation is an essential research to reveal the triggering mechanism of flesh quality deterioration. This study was aimed at explaining apoptotic mechanism of postmortem fish in terms of caspases activation, cytochrome c (cyt-c) release, B-cell lymphoma 2 (Bcl-2) and Bcl2-associated X (Bax) protein levels, transcriptional levels of its molecules, and apoptosis-inducing factor (AIF) translocation at 4 °C for 5 days. RESULTS: Activation of caspase-9, caspase-8, caspase-3 and the release of mitochondrial cyt-c were observed during storage. The decreased Bcl-2 protein levels, increased Bax protein expressions and Bax/Bcl-2 ratio were major steps for inducing apoptosis. Collectively, transcriptional regulation of Fas ligand (FasL), apoptotic protease activating factor-1 (Apaf-1), inhibitors of apoptosis proteins (IAPs), myeloid cell leukemia-1 (Mcl-1), c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) indicated that extrinsic apoptotic pathways (FasL/caspase-8/caspase-3) and intrinsic pathway [(JNK and p38 MAPK)/(Bcl-2, Bax and Mcl-1)/cyt-c/Apaf-1/caspase-9/caspase-3] were involved in apoptotic process. Mitochondrial AIF translocation to nuclear indicated that AIF mediated caspase-independent pathway. CONCLUSION: Therefore, transcriptional and translational alterations of multiple signaling molecules acted important roles in regulating apoptosis activation in postmortem process. © 2022 Society of Chemical Industry.


Assuntos
Carpas , Animais , Carpas/genética , Carpas/metabolismo , Proteína X Associada a bcl-2/metabolismo , Caspase 9/metabolismo , Caspase 8/metabolismo , Caspase 3/metabolismo , Proteína de Sequência 1 de Leucemia de Células Mieloides/metabolismo , Apoptose , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Músculos/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo
3.
Dev Comp Immunol ; 138: 104516, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36084755

RESUMO

Studies on host immunity evasion by aquatic viruses have largely focused on coding genes. There is accumulating evidence for the important biological functions of non-coding miRNAs in virus-host interactions. The regulatory functions of non-coding miRNAs in fish reovirus-host interactions remain unknown. Here, miR-2188-5p in grass carp (Ctenopharyngodon idellus), a miRNA specific to teleosts, was predicted to target the 3' UTR of the transcription factor klf2a. A correlation analysis and dual-luciferase reporter assay revealed that miR-2188-5p could induce the degradation of klf2a. The expression of miR-2188-5p induced the degradation of klf2a in a dose-dependent manner, suppressing the type I interferon response and promoting grass carp reovirus (GCRV) replication. As determined by a co-expression analysis, klf2a inhibited viral infection when miR-2188-5p was overexpressed. The targeted degradation of klf2a by miR-2188-5p could inhibit the type I interferon response and promote the replication of GCRV; however, this targeted degradation ability was insufficient to fully inhibit GCRV infection. These results provide novel insights into the regulatory effects and biological functions of non-coding miRNAs in fish-virus interactions.


Assuntos
Carpas , Doenças dos Peixes , Interferon Tipo I , MicroRNAs , Infecções por Reoviridae , Reoviridae , Regiões 3' não Traduzidas , Animais , Carpas/genética , Carpas/metabolismo , Interferon Tipo I/metabolismo , MicroRNAs/genética , Reoviridae/fisiologia , Fatores de Transcrição/genética
4.
Sci Total Environ ; 856(Pt 1): 159054, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36170916

RESUMO

Olanzapine (OLA) is a common drug used to treat schizophrenia and has recently come under increasing scrutiny as an emerging contaminant. However, its impact on lipid metabolism in fish and its mechanisms of action are not well understood. In this study, common carp were exposed to 0, 10, 100, and 250 µM OLA for 60 days. The results indicated that OLA exposure increased weight gain, total cholesterol (TC), low-density lipoprotein (LDL), and triglycerides (TG) and decreased high-density lipoprotein (HDL). In addition, lipids accumulated in the liver of the common carp. To explore the underlying mechanisms of action, gut microbiota, short-chain fatty acids (SCFAs), liver transcripts, and genes related to lipid metabolism were measured. It was discovered that OLA exposure altered the common carp gut microbiota composition and increased the abundance of SCFA-producing bacteria. Correspondingly, this study showed that OLA exposure increased the levels of SCFAs, which are highly relevant to the development of lipid accumulation. Transcriptome sequencing results indicated that OLA exposure could change lipid metabolism signalling pathways, including steroid biosynthesis, the PPAR signalling pathway, asglycerophospholipid metabolism, glycerolipid metabolism, and fatty acid metabolic pathways of the common carp. Additionally, OLA exposure interrupted lipid metabolism by means of significant upregulation of lipid synthesis-related genes, including pparγ, srebp1, and fas. OLA exposure also resulted in significant lipolysis-related gene downregulation, including cpt, lpl, hsl, and pparα. The results of this study indicated that contamination of aquatic environments with OLA alters lipid metabolism in common carp. In addition, the underlying mechanism might be due in part to the modulation of the gut microbiota-SCFA-PPAR signalling pathway.


Assuntos
Antipsicóticos , Carpas , Microbioma Gastrointestinal , Animais , Carpas/metabolismo , Metabolismo dos Lipídeos , Olanzapina/metabolismo , Antipsicóticos/metabolismo , Receptores Ativados por Proliferador de Peroxissomo/metabolismo , Ácidos Graxos Voláteis/metabolismo , Fígado/metabolismo
5.
Artigo em Inglês | MEDLINE | ID: mdl-36283647

RESUMO

Among pesticides, fungicides are the most extensively used ones in agriculture and their effects on fish health and indirectly human health are needed to evaluate. Folpet has been used for over 50 years as a fungicide across the world. The mechanism of action of folpet on non-target aquatic organisms is so poorly understood and there is no available information about potential acute toxicity of folpet and its mechanism of action in non-target aquatic organisms. With this motivation, two successive experiments were set up: first, 96 h-LC50 value of folpet for common carp (Cyprinus carpio) was determined; and second, effects of different sublethal concentrations (0, 0.025, 0.50, 0.1 and 0.15 mg/L) of folpet on hematological serum biochemical blood parameters, DNA damages, expression levels of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione S-transferase (GST), and cortisol receptor (HSP70) genes were evaluated in this study. It was calculated that 0.199 mg/L of folpet was the 96 h-LC50 value. After the folpet exposure for 14 days, significant decreases in red blood cells, hemoglobin, hematocrit, serum total protein, and GST gene expression levels were observed while serum glucose, liver enzymes activities, and expression levels of CAT, SOD, GPx, HSP70 genes increased. Also, folpet induced a significant genotoxic effect on the blood cells regarding to DNA damages. Consequently, the results have shown the toxic effects of folpet even at the lowest concentration on common carp.


Assuntos
Carpas , Fungicidas Industriais , Poluentes Químicos da Água , Animais , Humanos , Carpas/metabolismo , Fungicidas Industriais/toxicidade , Estresse Oxidativo , Poluentes Químicos da Água/toxicidade , Catalase/metabolismo , Glutationa Peroxidase/metabolismo , Superóxido Dismutase/metabolismo , Antioxidantes/metabolismo
6.
Food Chem ; 398: 133905, 2023 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-35969991

RESUMO

Maillard reaction products (MRPs) play pivotal roles in gut health by affecting the microbiome-host interactions. This study aimed at investigating the effects of MRPs derived from bighead carp meat hydrolysates with galactose and galacto-oligosaccharides on intestinal microbial composition and metabolic profile by in vitro pig fecal fermentation. The pH decreased sharply in the first 12 h and the highest production of butyric acid was observed in GM (glycated BCH with galacto-oligosaccharide) treatment with 64.7 µmoL/10 mL (p < 0.05) at 48 h. Clostridium_sensu_stricto_1, Streptococcus, and Enterococcus were dominant in the GM treatment, while Escherichia-Shigella was predominant in LgM (glycated BCH with galactose) treatment at 12 h. The up-regulated metabolites indicated that GM and LgM might participate in the fatty acids synthesis and modulate lipid metabolism, respectively. Overall, GM will be more beneficial for gut health by promoting the production of butyric acid and fatty acids synthesis.


Assuntos
Carpas , Microbioma Gastrointestinal , Animais , Ácido Butírico , Carpas/metabolismo , Ácidos Graxos Voláteis/metabolismo , Fezes/química , Fermentação , Galactose/análise , Produtos Finais de Glicação Avançada/análise , Carne , Metaboloma , Oligossacarídeos/química , Suínos
7.
Food Chem ; 399: 133799, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-35998490

RESUMO

Flesh quality is evaluated according to nutritional value and sensory quality. Cinnamaldehyde (CIN) improves mammalian meat quality, but research relating this to aquaculture is scarce. In this study, five doses of CIN (0, 36, 72, 108, 144 mg/kg diet) were fed to grass carp (Ctenopharyngodon idella) for 60 days. The results show that CIN supplementation increased nutritional value by increasing crude protein content. CIN also improved the sensory quality by increasing the pH and collagen content, decreasing shear force, lactate, and cooking loss. These changes may be related to changes in muscle fiber growth by increasing myofiber diameter. The increased myofiber diameter induced by CIN is associated with TOR mRNA and protein levels, and down-regulated FOXO3a mRNA levels, which might be associated with PTP1B/IGF1/PI3K/AKTs-TOR/FOXO3a signaling. Based on muscle crude protein content, optimal CIN supplementation dosage was 88.01 mg/kg.


Assuntos
Carpas , Doenças dos Peixes , Acroleína/análogos & derivados , Ração Animal/análise , Animais , Carpas/genética , Carpas/metabolismo , Dieta , Suplementos Nutricionais , Doenças dos Peixes/genética , Proteínas de Peixes/metabolismo , Imunidade Inata , Mamíferos/genética , Fibras Musculares Esqueléticas/metabolismo , Proteínas Musculares/metabolismo , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , RNA Mensageiro/genética , Transdução de Sinais
8.
Front Immunol ; 13: 1019872, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36330521

RESUMO

The common carp (Cyprinus carpio L.) is an important farmed species worldwide. Mucosal-associated lymphoid tissues play an essential role in the fight against pathogen infection. Spring viremia of carp virus (SVCV) poses a serious threat to the common carp aquaculture industry. Understanding the molecular mechanisms driving mucosal immune responses to SVCV infection is critical. In this study, the mucosal tissues (gills, foregut and hindgut) were collected from normal and infected fishes for transcriptome analysis. A total of 932,378,600 clean reads were obtained, of which approximately 80% were successfully mapped to the common carp genome. 577, 1,054 and 1,014 differential expressed genes (DEGs) were identified in the gills, foregut and hindgut, respectively. A quantitative polymerase chain reaction assay indicated that the DEGs expression in the foregut following SVCV infection was consistent with the transcriptome results. Among them, two key genes of the retinoic acid-inducible gene I (RIG-I)-like receptor family, melanoma-differentiation-associated gene 5 (MDA5) and laboratory of genetics and physiology 2 (LGP2) (i.e., CcMDA5 and CcLGP2), underwent further analysis. Overexpression of CcMDA5 or CcLGP2 increased phosphorylation of TANK-binding kinase 1 and interferon regulatory factor 3 and the expression of interferon-1 (ifn-1), myxovirus resistance (mx), viperin and interferon-stimulated gene 15 (isg15), and inhibited SVCV replication in epithelioma papulosum cyprini cells. Furthermore, CcLGP2 significantly upregulated the CcMDA5-induced ifn-1 mRNA expression and the activation of the ifn-1 promoter. Finally, confocal microscopy and coimmunoprecipitation experiments revealed that CcLGP2 colocalizes and interacts with CcMDA5 via the C-terminal regulatory domain. This study provides essential gene resources for understanding the fish immune response to SVCV infection and sheds light on the potential role of fish LGP2 in the MDA5 regulation.


Assuntos
Carpas , Doenças dos Peixes , Infecções por Rhabdoviridae , Animais , Carpas/genética , Carpas/metabolismo , Viremia , Interferons/metabolismo , Perfilação da Expressão Gênica , Antivirais
9.
Front Immunol ; 13: 965954, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36405693

RESUMO

High-fat diet is regarded as crucial inducers of oxidative stress, inflammation, and metabolic imbalance. In order to investigate the ameliorative potential of resveratrol against the progression of liver injury towards steatohepatitis, common carp (Cyprinus carpio) were distributed into six experimental groups and were fed with a normal-fat diet, a high-fat diet, and supplemented with resveratrol (0.8, 1.6, 2.4, and 3.2 g/kg diet) for 8 weeks. The high-fat diet decreased the antioxidant capacities, as well as causing the inflammatory response and lipid deposition of common carp. Resveratrol induced a marked elevation in the final body weight, weight gain rate, condition factor and significant decrease in the feed conversion ratio. Moreover, dietary resveratrol showed a significant decrease in the alanine aminotransferase, aspartate aminotransferase, triglyceride and low-density lipoprotein levels, which was accompanied by an increase in high-density lipoprotein concentration in serum. A significant elevation in total superoxide dismutase, catalase, glutathione peroxidase and a decreased malondialdehyde content were observed, along with a substantial elevation in antioxidant activities were found. Additionally, fish fed with resveratrol had an up-regulation of hepatic catalase, copper, zinc superoxide dismutase, glutathione peroxidase 1a, and glutathione peroxidase 1b gene expression via Nrf2 signaling pathway. Expectedly, our results also demonstrated that resveratrol regulates hepatic lipid metabolism in fish by inhibiting the expression of hepatic lipogenesis genes (acetyl-CoA carboxylase 1, fatty acid synthase, and sterol regulatory element binding protein 1), fatty acid uptake-related genes of lipoprotein lipase, and ß-oxidation-related genes via PPAR-γ signaling pathway. Furthermore, dietary resveratrol reduced inflammation, as evident by down-regulating the interleukin-1ß, interleukin-6, interleukin-8, and tumor necrosis factor-α expression levels and upregulating the interleukin-10 and transforming growth factor-ß2 expression levels via NF-κB signaling pathway. As a whole, our results demonstrated that resveratrol defensed the impacts against high-fat diet on the serum biochemical, hepatic antioxidants, inflammation, and lipid metabolism.


Assuntos
Carpas , Dieta Hiperlipídica , Animais , Dieta Hiperlipídica/efeitos adversos , Metabolismo dos Lipídeos/genética , Resveratrol/farmacologia , Carpas/metabolismo , Catalase/metabolismo , Antioxidantes/farmacologia , Antioxidantes/metabolismo , Glutationa Peroxidase/metabolismo , Estresse Oxidativo , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Superóxido Dismutase/metabolismo
10.
J Immunol ; 209(10): 1918-1929, 2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-36198423

RESUMO

Cyprinid herpesvirus 3 (CyHV-3) has caused severe economic losses to carp culture, but its pathogenicity is far from clear. Our previous study has revealed that microRNA (miR)-722 was upregulated during CyHV-3 infection, indicating that miR-722 might play an important role in CyHV-3 replication. In this study, we found that overexpression of miR-722 inhibited CyHV-3 replication and promoted IFN expression. The putative target gene of miR-722 was searched over the CyHV-3 genome, and ORF89 was identified and validated as a target gene of miR-722. Overexpression of ORF89 markedly reduced the expression of IFN and IFN-stimulated genes. Mechanistically, ORF89 interacted with and degraded IFN regulatory factor 3 (IRF3), and inhibited the entry of IRF3 into the nucleus by suppressing the dimerization of IRF3. Moreover, ORF89-mediated suppression of IFN expression could be restored by adding miR-722. To our knowledge, our findings confirm a novel virus-host combat, in which CyHV-3 evades host antiviral immunity by its ORF89 protein, whereas host miR-722, upregulated on CyHV-3 infection, targets ORF89 to impede CyHV-3 replication.


Assuntos
Carpas , Doenças dos Peixes , Infecções por Herpesviridae , Herpesviridae , MicroRNAs , Animais , Fator Regulador 3 de Interferon/genética , Fator Regulador 3 de Interferon/metabolismo , Evasão da Resposta Imune , Proteínas Virais/genética , Proteínas Virais/metabolismo , MicroRNAs/genética , Herpesviridae/genética , Carpas/genética , Carpas/metabolismo , Replicação Viral
11.
Ecotoxicol Environ Saf ; 246: 114136, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36242823

RESUMO

Copper (Cu) is a common contaminant in aquatic environments, which could cause physiological dysfunction in aquatic organisms. However, few studies have comprehensively examined the impact of copper toxicity in freshwater fish over the past decade. In this research, the oxidative stress, liver transcriptome, intestinal microbiota, and histopathology of common carp (C. carpio) in response to Cu exposure were studied, by exposing juvenile carp to 0.2 mg/ml Cu2+ for 30 days. The results revealed that Cu2+ could induce significant changes in malondialdehyde (MDA) content and antioxidant enzyme (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx)) activity. The changes in antioxidant enzyme activities indicate that Cu can induce oxidative stress by generating reactive oxygen species (ROS) content. RNA-seq analysis of the liver identified 1069 differentially expressed genes (DEGs) after treatment with 2.0 mg/L Cu2+. Among the DEGs, 490 genes were upregulated and 579 genes were downregulated. GO functional enrichment analysis revealed that Cu could affect the fatty acid biosynthetic process, carnitine biosynthetic process, and activity of carboxylic acid transmembrane transporter. Meanwhile, the most significantly enriched KEGG pathway also included the lipid metabolism pathway. In addition, Cu2+ exposure increased bacterial richness and changed bacterial composition. At the phylum level, we found that the ratio of Bacteroidetes to Firmicutes was increased in the treatment carps, which can regulate intestinal epithelium function and reduce inflammation and immune responses. At the genus level, the abundances of 11 genera were significantly altered after exposure to Cu2+. The altered composition of the microbial community caused by Cu exposure may play a useful role in compensation of the intestinal lesions by Cu exposure. Furthermore, we found that Cu2+ exposure could cause histological alterations such as structural damage to the liver and intestines. The results of this research contribute to a better understanding of mechanisms related to Cu toxicity in fish.


Assuntos
Carpas , Microbioma Gastrointestinal , Poluentes Químicos da Água , Animais , Carpas/genética , Carpas/metabolismo , Cobre/toxicidade , Antioxidantes/metabolismo , Transcriptoma , Estresse Oxidativo , Poluentes Químicos da Água/toxicidade
12.
Oxid Med Cell Longev ; 2022: 4972622, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36267815

RESUMO

Salinization of aquatic ecosystem, abrupt climate change, and anthropogenic activities cause adverse impact on agricultural land/soil as well as the aquaculture industry. This experimental study was designed to evaluate different biomarkers of oxidative stress, antioxidant enzymes, and genotoxic potential of diverse salinities of brackish water on freshwater fish. A total of 84 fresh water mrigal carp (Cirrhinus mrigala) were randomly segregated and maintained in four groups (T0, T1, T2, and T3) in a glass aquarium under similar laboratory conditions at various salinity levels (0, 3, 5, and 7 parts per thousand) to determine the pathological influence of brackish water. All the fish in groups T1, T2, and T3 were exposed to various salinity levels of brackish water for a period of 90 days while the fish of group T0 served as the control group. The experimental fish reared in different groups T1, T2, and T3 displayed various physical and behavioral ailments. The results revealed significantly augmented quantity of different oxidative stress indicators including reactive oxygen species (ROS) and thiobarbituric acid reactive substance (TBARS) in different visceral tissues (kidneys, liver, and gills) of exposed fish. Different antioxidant enzymes such as reduced glutathione (GSH), peroxidase (POD), superoxide dismutase (SOD), and catalase (CAT) along with total proteins were remarkably reduced in the kidneys, gills, and liver tissues. Results showed significantly increased values of different nuclear abnormalities (erythrocyte with micronucleus, erythrocyte with condensed nucleus, and erythrocyte with lobed nucleus) and morphological changes (pear shaped erythrocyte, spindle-shaped erythrocytes, and spherocyte) in red blood cells of experimental fish. The results on genotoxic effects exhibited significantly increased DNA damage in isolated cells of liver, kidneys, and gills of exposed fish. The findings of our experimental research suggested that brackish water causes adverse toxicological impacts on different visceral tissues of fresh water fish at higher salinity level through disruption and disorder of physiological and biochemical markers.


Assuntos
Carpas , Poluentes Químicos da Água , Animais , Catalase/metabolismo , Antioxidantes/farmacologia , Carpas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Ecossistema , Poluentes Químicos da Água/toxicidade , Superóxido Dismutase/metabolismo , Estresse Oxidativo , Glutationa/metabolismo , Eritrócitos/metabolismo , Fígado/metabolismo , Águas Salinas , Biomarcadores/metabolismo , Solo
13.
Int J Mol Sci ; 23(19)2022 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-36232671

RESUMO

Complement factor I (CFI), a complement inhibitor, is well known for regulating the complement system activation by degrading complement component 3b (C3b) in animal serum, thus becoming involved in innate defense. Nevertheless, the functional mechanisms of CFI in the complement system and in host-pathogen interactions are far from being clarified in teleost fish. In the present study, we cloned and characterized the CFI gene, CiCFI, from grass carp (Ctenopharyngodon idella) and analyzed its function in degrading serum C3b and expression changes after grass carp reovirus (GCRV) infection. The open reading frame of CiCFI was found to be 2121 bp, encoding 706 amino acids with a molecular mass of 79.06 kDa. The pairwise alignments showed that CiCFI shared the highest identity (66.9%) with CFI from Carassius gibelio and the highest similarity (78.7%) with CFI from Danio rerio. The CiCFI protein was characterized by a conserved functional core Tryp_SPc domain with the catalytic triad and substrate binding sites. Phylogenetic analysis indicated that CiCFI and the homologs CFIs from other teleost fish formed a distinct evolutionary branch. Similar with the CFIs reported in mammals, the recombinant CiCFI protein could significantly reduce the C3b content in the serum, demonstrating the conserved function of CiCFI in the complement system in the grass carp. CiCFI mRNA and protein showed the highest expression level in the liver. After GCRV infection, the mRNA expressions of CiCFI were first down-regulated, then up-regulated, and then down-regulated to the initial level, while the protein expression levels maintained an overall downward trend to the late stage of infection in the liver of grass carps. Unexpectedly, the protein levels of CiCFI were also continuously down-regulated in the serum of grass carps during GCRV infection, while the content of serum C3b proteins first increases and then returns to the initial level, suggesting a distinct role of CiCFI in regulating complement activation and fish-virus interaction. Combining our previous results that complement factor D, a complement enhancer, shows continuously up-regulated expression levels in grass carps during GCRV infection, and this study may provide the further essential data for the full picture of complex complement regulation mechanism mediated by Df and CFI of the grass carp during pathogen infection.


Assuntos
Carpas , Doenças dos Peixes , Infecções por Reoviridae , Reoviridae , Aminoácidos/metabolismo , Animais , Carpas/genética , Carpas/metabolismo , Ativação do Complemento , Complemento C3b , Fator D do Complemento/genética , Fator I do Complemento/genética , Fator I do Complemento/metabolismo , Inativadores do Complemento , Proteínas de Peixes/metabolismo , Regulação da Expressão Gênica , Mamíferos/metabolismo , Filogenia , RNA Mensageiro/genética , Reoviridae/fisiologia , Infecções por Reoviridae/genética , Infecções por Reoviridae/veterinária
14.
Int J Mol Sci ; 23(19)2022 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-36232849

RESUMO

A high-quality baseline transcriptome is a valuable resource for developmental research as well as a useful reference for other studies. We gathered 41 samples representing 11 tissues/organs from 22 important developmental time points within 197 days of fertilization of grass carp eggs in order to systematically examine the role of lncRNAs and alternative splicing in fish development. We created a high-quality grass carp baseline transcriptome with a completeness of up to 93.98 percent by combining strand-specific RNA sequencing and single-molecule real-time RNA sequencing technologies, and we obtained temporal expression profiles of 33,055 genes and 77,582 transcripts during development and tissue differentiation. A family of short interspersed elements was preferentially expressed at the early stage of zygotic activation in grass carp, and its possible regulatory components were discovered through analysis. Additionally, after thoroughly analyzing alternative splicing events, we discovered that retained intron (RI) alternative splicing events change significantly in both zygotic activation and tissue differentiation. During zygotic activation, we also revealed the precise regulatory characteristics of the underlying functional RI events.


Assuntos
Carpas , Doenças dos Peixes , RNA Longo não Codificante , Processamento Alternativo , Animais , Carpas/genética , Carpas/metabolismo , Doenças dos Peixes/genética , Proteínas de Peixes/genética , Perfilação da Expressão Gênica , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Transcriptoma
15.
Int J Mol Sci ; 23(19)2022 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-36233179

RESUMO

Gelsemium elegans&nbsp;Benth. (GEB) is a traditional medicinal plant in China, and acts as a growth promoter in pigs and goats. Koumine (KM) is the most abundant alkaloid in GEB and produces analgesic, anti-cancer, and immunomodulatory effects. KM can be used as an aquatic immune stimulant, but its growth-promoting effects and transcriptional mechanisms have not been investigated. Diets containing KM at 0, 0.2, 2, and 20 mg/kg were fed to Cyprinus carpio for 71 days to investigate its effects on growth performance, intestinal morphology, microflora, biochemical indicators, and transcriptional mechanisms. Cyprinus carpio fed with KM as the growth promoter, and the number of intestinal crypts and intestinal microbial populations were influenced by KM concentration. KM increased the abundance of colonies of Afipia, Phyllobacterium, Mesorhizobium, and Labrys, which were associated with compound decomposition and proliferation, and decreased the abundance of colonies of pathogenic bacteria Methylobacterium-Methylorubrum. A total of 376 differentially-expressed genes (DEGs) among the four experimental groups were enriched for transforming growth factor-ß1 and small mother against decapentaplegic (TGF-ß1/Smad), mitogen-activated protein kinase (MAPK), and janus kinases and signal transducers and activators of transcription (Jak/Stat) signaling pathways. In particular, tgfbr1, acvr1l, rreb-1, stat5b, smad4, cbp, and c-fos were up-regulated and positively correlated with KM dose. KM had a growth-promoting effect that was related to cell proliferation driven by the TGF-ß1/Smad, MAPK, and Jak/Stat signaling pathways. KM at 0.2 mg/kg optimized the growth performance of C. carpio, while higher concentrations of KM (2 and 20 mg/kg) may induce apoptosis without significantly damaging the fish intestinal structure. Therefore, KM at low concentration has great potential for development as an aquatic growth promotion additive.


Assuntos
Carpas , Microbiota , Ração Animal/análise , Animais , Carpas/genética , Carpas/metabolismo , Dieta/veterinária , Suplementos Nutricionais/análise , Alcaloides Indólicos , Janus Quinases , Proteínas Quinases Ativadas por Mitógeno , Receptor do Fator de Crescimento Transformador beta Tipo I , Suínos , Fator de Crescimento Transformador beta1/metabolismo
16.
Front Endocrinol (Lausanne) ; 13: 982297, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36303873

RESUMO

Gonadotropin-releasing hormone (GnRH), as a vital hypothalamic neuropeptide, was a key regulator for pituitary luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in the vertebrate. However, little is known about the other pituitary actions of GnRH in teleost. In the present study, two GnRH variants (namely, GnRH2 and GnRH3) and four GnRH receptors (namely, GnRHR1, GnRHR2, GnRHR3, and GnRHR4) had been isolated from grass carp. Tissue distribution displayed that GnRHR4 was more highly detected in the pituitary than the other three GnRHRs. Interestingly, ligand-receptor selectivity showed that GnRHR4 displayed a similar and high binding affinity for grass carp GnRH2 and GnRH3. Using primary culture grass carp pituitary cells as model, we found that both GnRH2 and GnRH3 could not only significantly induce pituitary reproductive hormone gene (GtHα, LHß, FSHß, INHBa, secretogranin-2) mRNA expression mediated by AC/PKA, PLC/IP3/PKC, and Ca2+/CaM/CaMK-II pathways but also reduce dopamine receptor 2 (DRD2) mRNA expression via the Ca2+/CaM/CaMK-II pathway. Interestingly, GnRH2 and GnRH3 could also stimulate anorexigenic peptide (POMCb, CART2, UTS1, NMBa, and NMBb) mRNA expression via AC/PKA, PLC/IP3/PKC, and Ca2+/CaM/CaMK-II pathways in grass carp pituitary cells. In addition, food intake could significantly induce brain GnRH2 mRNA expression. These results indicated that GnRH should be the coupling factor to integrate the feeding metabolism and reproduction in teleost.


Assuntos
Carpas , Hormônio Liberador de Gonadotropina , Animais , Hormônio Liberador de Gonadotropina/metabolismo , Hipófise/metabolismo , Reprodução/fisiologia , Hipotálamo/metabolismo , Hormônios Hipofisários , Carpas/metabolismo , RNA Mensageiro/genética
17.
Cells ; 11(20)2022 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-36291067

RESUMO

The number of metabolic syndromes (MetS) is increasing, and a fish phospholipid diet can reduce the risk of MetS. In this study, the changes in lipid metabolism of colon contents were analyzed by extensive lipidomics in mice with metabolic syndrome by fish phospholipid diet, and mice were randomly divided into experimental groups with different diet types by establishing a MetS model. After 14 weeks, the mice were sacrificed and the serum and colon contents were collected. Ultra-high liquid phase tandem mass spectrometry was used for broadly targeted lipidomic analysis, and the qualitative and quantitative detection of lipid metabolism changes in the colonic contents of mice. Under the intervention of fish phospholipids, MetS mice were significantly inhibited, serum total cholesterol (TC) and triglycerides (TG) decreased, serum high-density lipoprotein (HDL-C) and low-density lipoprotein (LDL-C) levels were improved, fasting blood glucose and insulin levels decreased, and inflammatory factors decreased. Through screening, it was found that thirty-three lipid metabolites may be key metabolites and five have significantly changed metabolic pathways. Modularizing lipid metabolites, it is possible to understand the extent to which different types and concentrations of fish phospholipids affect metabolic syndrome. Therefore, our study may provide new therapeutic clues for improving MetS.


Assuntos
Carpas , Insulinas , Síndrome Metabólica , Camundongos , Animais , Fosfolipídeos , Lipidômica , Glicemia/metabolismo , LDL-Colesterol , Salmão/metabolismo , Carpas/metabolismo , Triglicerídeos , Lipoproteínas HDL
18.
Artigo em Inglês | MEDLINE | ID: mdl-36108998

RESUMO

Lead (Pb) is one of the most common trace metals in water, and its high concentration in the environment can cause harm to aquatic animals and humans. In the present study, the effects of Pb exposure (3.84 mg/kg) on the morphology, digestive enzyme activity, immune function and microbiota structure of silver carp (Hypophthalmichthys molitrix) intestines within 96 h were detected. Moreover, the correlation between them was analyzed. The results showed that Pb exposure on the one hand severely impaired the intestinal morphology, including significantly shortening the intestinal villi's length, increasing the goblet cells' number, causing the intestinal leukocyte infiltration, and thickening the intestinal wall abnormally, on the other hand, increasing the activity of intestinal digestive enzyme (trypsin and lipase). In addition, the mRNA expressions of structure-related genes (Claudin-7 and villin-1) were down-regulated, and the immune factors genes (IL-8, IL-10 and TNF-α) were up-regulated after Pb exposure. Furthermore, data of the MiSeq sequencing showed that the abundance of membrane transport, immune system function and digestive system of silver carp intestinal microbiota all decreased, while cellular antigens increased. Finally, the canonical correlation analysis (CCA) showed that there were correlations between silver carp's intestinal microbiota and intestinal morphology and immune factors. In conclusion, it is speculated that the entry of Pb into the intestine leads the microbiota dysbiosis, affects the intestinal immunity and digestive function, and further damages the intestinal barrier of silver carp.


Assuntos
Carpas , Microbioma Gastrointestinal , Animais , Carpas/metabolismo , Claudinas , Disbiose/induzido quimicamente , Disbiose/veterinária , Humanos , Imunidade , Interleucina-10 , Interleucina-8 , Intestinos , Chumbo/toxicidade , Lipase , RNA Mensageiro/metabolismo , Tripsina , Fator de Necrose Tumoral alfa , Água
19.
Fish Shellfish Immunol ; 130: 114-131, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36084887

RESUMO

Chemokines are a group of secreted small molecules which are essential for cell migration in physiological and pathological conditions by binding to specific chemokine receptors. They are structurally classified into five groups, namely CXC, CC, CX3C, XC and CX. CC chemokine group is the largest one among them. In this study, we identified and characterized 61 CC chemokines from allotetraploid common carp (Cyprinus carpio). The sequence analyses showed that the majority of CC chemokines had an N-terminal signal peptide, and an SCY domain, and all CC chemokines were located in the extracellular region. Phylogenetic, evolutionary and syntenic analyses confirmed that CC chemokines were annotated as 11 different types (CCL19, CCL20, CCL25, CCL27, CCL32, CCL33, CCL34, CCL35, CCL36, CCL39, and CCL44), which exhibited unique gene arrangement pattern and chromosomal location respectively. Furthermore, genome synteny analyses between common carp and four representative teleost species indicated expansion of common carp CC chemokines resulted from the whole genome duplication (WGD) event. Additionally, the continuous evolution of gene CCL25s in teleost afforded a novel viewpoint to explain the WGD event in teleost. Then, we predicted the three-dimensional structures and probable function regions of common carp CC chemokines. All the CC chemokines core structures were constituted of an N-loop, a three-stranded ß-sheet, and a C-terminal helix. Finally, 43 CC chemokines were predicted to have probable general antimicrobial activity. Their tertiary structures, cationic and amphiphilic physicochemical property supported the viewpoint. To verify the prediction, six recombinant CCL19s proteins were prepared and the antibacterial activity against Escherichia coli and Aeromonas hydrophila were verified. The results supported our prediction that rCCL19a.1s (rCCL19a.1_a, rCCL19a.1_b) and rCCL19bs (rCCL19b_a, rCCL19b_b), especially rCCL19bs, exhibited extremely significant inhibition to the growth of both E. coli and A. hydrophila. On the contrary, two rCCL19a.2s had no significant inhibitory effect. These studies suggested that CC chemokines were essential in immune system evolution and not monofunctional during pathogen infection.


Assuntos
Carpas , Animais , Antibacterianos , Carpas/genética , Carpas/metabolismo , Quimiocinas/genética , Quimiocinas CC/genética , Escherichia coli , Proteínas de Peixes , Filogenia , Sinais Direcionadores de Proteínas/genética , Receptores de Quimiocinas/genética
20.
Fish Shellfish Immunol ; 130: 332-341, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36115605

RESUMO

Grass carp (Ctenopharyngodon idella) is the largest economic fish in freshwater culture in China, which is predisposed to infectious diseases under high temperature. Under the background of global warming, the industrialization of the Pearl River Delta region has led to aggravated thermal pollution, which has increasingly serious impacts on the aquatic ecological environment. This will result in more frequent exposure of grass carp to overheated water temperatures. Previous studies have only identified the regulatory genes of fish that respond to pathogens or temperature stress, but the transcriptional response to both is unknown. In this study, the histopathological analysis showed heat stress exacerbated spleen damage induced by Aeromonas hydrophila. The transcriptional responses of the spleens from A. hydrophila lipopolysaccharide (LPS) -injected grass carp undergoing heat stress and at normal temperatures for 6, 24, and 72 h were investigated by mRNA and microRNA sequencing. We identified 28, 20, and 141 differentially expressed (DE) miRNAs and 126, 383, and 4841 DE mRNAs between the two groups after 6, 24, and 72 h, respectively. There were 67 DE genes mainly involved in the cytochrome P450 pathway, antioxidant defense, inflammatory response, pathogen recognition pathway, antigen processing and presentation, and the ubiquitin-proteasome system. There were 5 DE miRNAs involved in regulating apoptosis and inflammation. We further verified 17 DE mRNAs and 5 DE miRNAs using quantitative real-time PCR. Based on miRNAs and mRNAs analysis, continuous heat stress will affect the antibacterial responses of grass carp spleens, resulting in aggravation of spleen injury. Together, these results provide data for further understanding of the decreased tolerance of fish to pathogen infection in persistent high-temperature environments.


Assuntos
Carpas , Doenças dos Peixes , Infecções por Bactérias Gram-Negativas , MicroRNAs , Aeromonas hydrophila/fisiologia , Animais , Antibacterianos , Antioxidantes , Carpas/genética , Carpas/metabolismo , Proteínas de Peixes , Resposta ao Choque Térmico , Imunidade Inata/genética , Lipopolissacarídeos/farmacologia , MicroRNAs/genética , Complexo de Endopeptidases do Proteassoma , RNA Mensageiro/metabolismo , Ubiquitinas , Água
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