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1.
Gene ; 737: 144433, 2020 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-32014563

RESUMO

The Carassius auratus (crucian carp) complex of the Dongting water system exhibits coexistence of diploid and triploid forms. As reported, triploid C. auratus is autotriploid origin. Ribosomal DNA (rDNA) with evolutionary conservation is widely used to study polyploidization. Here, we investigated genomic and transcribed rDNA sequences (18S and 5S) in diploid (2nCC, 2n = 100) and triploid (3nCC, 3n = 150) C. auratus. The results showed that the genetic traits and expression of 18S and 5S rDNA from 2nCC individuals were identified in 3nCC individuals. Moreover, pseudogenization of rDNA (18S and 5S) sequences were also observed in both 2nCC and 3nCC individuals, but expression of these variants was not detected. Based on the transcribed rDNA consensus sequence between 2nCC and 3nCC individuals, the functional secondary structures of 18S rRNA (expansion segments, ES6S) and 5S rRNA were predicted. These data demonstrated that complex evolutionary dynamics existed in the rDNA family of C. auratus. The evolutionary conservation of rDNA revealed that autotriploidization could not induce the divergence in Carassius taxa of the Dongting water system. These observations will expand our knowledge of the evolutionary dynamics of the rDNA family in vertebrates.


Assuntos
DNA Ribossômico/genética , Carpa Dourada/genética , RNA Ribossômico 18S/genética , RNA Ribossômico 5S/genética , Triploidia , Animais , Sequência de Bases , DNA Ribossômico/química , Evolução Molecular , Carpa Dourada/classificação , Conformação de Ácido Nucleico , Filogenia , RNA Ribossômico 18S/química , RNA Ribossômico 5S/química , Alinhamento de Sequência
2.
Mol Immunol ; 119: 18-26, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31954283

RESUMO

Fish interferon (IFN)-mediated antiviral innate immunity is the first line of defense against virus invasion. In the present study, we identify two fish IFN genes (here tentatively named IFNa and IFNc) with different-sized 3' UTRs from clone F strain of gibel carp Carassius auratus gibelio. Carp IFNa has a relatively short 3'UTR without AU-rich elements (AREs) but IFNc has a long one with 9 AREs. Functionally, carp IFNa and IFNc display significantly antiviral potential to viral infection, likely through induction of downstream IFN-stimulated genes (ISGs). Both carp IFN genes are induced by viral infection, poly(I:C) treatment and IRF3/7, which are ascribed to the IFN-sensitive response elements (ISRE) within their promoters. Carp IFN genes are also induced by each other and by themselves, indicating existence of a positive feedback loop in fish IFN-mediated antiviral immune response. Comparative analyses of 3'UTR-mediated expression regulation at mRNA and protein levels show that the ARE-containing 3'UTR of carp IFNc rather than the short 3'UTR of carp IFNa promotes mRNA decay but instead results in high-level protein expression, indicating that 3'UTR of fish IFN mRNAs might be a potential factor for regulation of IFN-mediated antiviral immune response. Considering a fact that a given protein function is largely related to its protein level, these results suggest that both promoter and 3'UTR contribute to the transcription and translation of fish IFN genes, thus shaping their eventually antiviral potential.


Assuntos
Regiões 3' não Traduzidas , Doenças dos Peixes/genética , Carpa Dourada/genética , Interferons/genética , Regulação para Cima , Elementos Ricos em Adenilato e Uridilato , Animais , Linhagem Celular , Doenças dos Peixes/imunologia , Infecções por Herpesviridae/genética , Infecções por Herpesviridae/veterinária , Infecções por Herpesviridae/virologia , Especificidade da Espécie
3.
Gen Comp Endocrinol ; 285: 113266, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31493394

RESUMO

In the present study, the effects of photic environments, such as background color (white and black) and chromatic lights (blue, green, and red), on body color and gene expressions of melanin-concentrating hormone (mch) in the brain and proopiomelanocortin (pomc) in the pituitary, as well as the roles of the eyes and brain as mediators of ambient light to these genes, were examined in goldfish (Carassius auratus). Body color of goldfish exposed to fluorescent light (FL) under white background (WBG) was paler than those under black background (BBG). Gene expression levels for mch and pomc were reciprocally different depending on background color; under WBG, mRNA levels of mch and pomc were high and low, respectively, while under BBG, these levels were reversed. mch and pomc mRNA expressions of the fish exposed to chromatic light from LED were primarily similar to those exposed to FL, while blue light stimulated the expressions of mch and pomc. Ophthalmectomized goldfish exposed to FL or blue light showed minimum expression levels of mch gene, suggesting that eyes are the major mediator of ambient light for mch gene expression. Contrastingly, mRNA expressions of pomc in ophthalmectomized goldfish exposed to FL were different from those of intact goldfish. These results suggest that eyes play a functional role in mediating ambient light to regulate pomc gene expression. Since ophthalmectomy caused an increase in pomc mRNA contents in the fish exposed to blue light, we suggest that the brain is an additional mediator to regulate pomc gene expression.


Assuntos
Regulação da Expressão Gênica , Carpa Dourada/genética , Hormônios Hipotalâmicos/genética , Luz , Melaninas/genética , Pigmentação/genética , Pigmentação/efeitos da radiação , Hormônios Hipofisários/genética , Pró-Opiomelanocortina/genética , Animais , Encéfalo/metabolismo , Encéfalo/efeitos da radiação , Cor , Regulação da Expressão Gênica/efeitos da radiação , Hormônios Hipotalâmicos/metabolismo , Melaninas/metabolismo , Hipófise/metabolismo , Hipófise/efeitos da radiação , Hormônios Hipofisários/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
4.
Genes (Basel) ; 10(9)2019 09 12.
Artigo em Inglês | MEDLINE | ID: mdl-31547242

RESUMO

In teleost, pigment in the skin and scales played important roles in various biological processes. Iridophores, one of the main pigment cells in teleost, could produce silver pigments to reflect light. However, the specific mechanism of the formation of silver pigments is still unclear. In our previous study, some transparent mutant individuals were found in the carp-goldfish nucleocytoplasmic hybrid (CyCa hybrid) population. In the present study, using transparent mutants (TM) and wild type (WT) of the CyCa hybrid as a model, firstly, microscopic observations showed that the silver pigments and melanin were both lost in the scales of transparent mutants compared to that in wild types. Secondly, genetic study demonstrated that the transparent trait in the CyCa hybrid was recessively inherent, and controlled by an allele in line with Mendelism. Thirdly, RNA-Seq analysis showed that differential expression genes (DEGs) between wild type and transparent mutants were mainly enriched in the metabolism of guanine, such as hydrolase, guanyl nucleotide binding, guanyl ribonucleotide binding, and GTPase activity. Among the DEGs, purine nucleoside phosphorylase 4a (pnp4a) and endothelin receptor B (ednrb) were more highly expressed in the wild type compared to the transparent mutant (p < 0.05). Finally, miRNA-Seq analysis showed that miRNA-146a and miR-153b were both more highly expressed in the transparent mutant compared to that in wild type (p < 0.05). Interaction analysis between miRNAs and mRNAs indicated that miRNA-146a was associated with six DEGs (MGAT5B, MFAP4, GP2, htt, Sema6b, Obscn) that might be involved in silver pigmentation.


Assuntos
Carpa Dourada/genética , Mutação , Pigmentação da Pele/genética , Animais , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Genes Recessivos , Guanina/metabolismo , Melaninas/genética , Melaninas/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Purina-Núcleosídeo Fosforilase/genética , Purina-Núcleosídeo Fosforilase/metabolismo , Receptores de Endotelina/genética , Receptores de Endotelina/metabolismo
5.
Fish Shellfish Immunol ; 94: 510-516, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31541778

RESUMO

Skin plays an important role in the innate immune responses of fish, particularly towards bacterial infection. To understand the molecular mechanism of mucosal immunity of fish during bacterial challenge, a de novo transcriptome assembly of crucian carp Carassius auratus skin upon Aeromonas hydrophila infection was performed, the latter with Illumina Hiseq 2000 platform. A total of 118111 unigenes were generated and of these, 9693 and 8580 genes were differentially expressed at 6 and 12 h post-infection, respectively. The validity of the transcriptome results of eleven representative genes was verified by quantitative real-time PCR (qRT-PCR) analysis. A comparison with the transcriptome profiling of zebrafish skin to A. hydrophila with regards to the mucosal immune responses revealed similarities in the complement system, chemokines, heat shock proteins and the acute-phase response. GO and KEGG enrichment pathway analyses displayed the significant immune responses included TLR, MAPK, JAK-STAT, phagosome and three infection-related pathways (ie., Salmonella, Vibrio cholerae and pathogenic Escherichia coli) in skin. To our knowledge, this study is the first to describe the transcriptome analysis of C. auratus skin during A. hydrophila infection. The outcome of this study contributed to the understanding of the mucosal defense mechanisms in cyprinid species.


Assuntos
Doenças dos Peixes/imunologia , Carpa Dourada/genética , Carpa Dourada/imunologia , Imunidade Inata/genética , Pele/metabolismo , Aeromonas hydrophila/fisiologia , Animais , Perfilação da Expressão Gênica/veterinária , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária
6.
Int J Mol Sci ; 20(16)2019 Aug 12.
Artigo em Inglês | MEDLINE | ID: mdl-31409051

RESUMO

It is now widely accepted that allele-specific DNA methylation (ASM) commonly occurs at non-imprinted loci. Most of the non-imprinted ASM regions observed both within and outside of the CpG island show a strong correlation with DNA polymorphisms. However, what polymorphic cis-acting elements mediate non-imprinted ASM of the CpG island remains unclear. In this study, we investigated the impact of polymorphic GT microsatellites within the gene promoter on non-imprinted ASM of the local CpG island in goldfish. We generated various goldfish heterozygotes, in which the length of GT microsatellites or some non-repetitive sequences in the promoter of no tail alleles was different. By examining the methylation status of the downstream CpG island in these heterozygotes, we found that polymorphisms of a long GT microsatellite can lead to the ASM of the downstream CpG island during oogenesis and embryogenesis, polymorphisms of short GT microsatellites and non-repetitive sequences in the promoter exhibited no significant effect on the methylation of the CpG island. We also observed that the ASM of the CpG island was associated with allele-specific expression in heterozygous embryos. These results suggest that a long polymorphic GT microsatellite within a gene promoter mediates non-imprinted ASM of the local CpG island in a goldfish inter-strain hybrid.


Assuntos
Metilação de DNA , Carpa Dourada/genética , Repetições de Microssatélites , Alelos , Animais , Quimera/genética , Ilhas de CpG , Cruzamentos Genéticos , Feminino , Impressão Genômica , Carpa Dourada/embriologia , Masculino , Polimorfismo Genético , Regiões Promotoras Genéticas
7.
Gen Comp Endocrinol ; 283: 113240, 2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-31394085

RESUMO

Leptin is primarily considered a peripheral satiety hormone and is also found to perform important roles in energy homeostasis in vertebrates ranging from fish to mammals. The liver is a major source of leptin production in teleost fish. Using goldfish as a model, a previous report by our group illustrated the positive regulation of leptin mRNA levels by treatment with the hyperglycemic hormone glucagon, and our present study provided evidence for the negative regulation of hepatic leptin-AI and leptin-AII transcripts through the administration of the hypoglycemic hormone insulin. This study is the first to demonstrate changes in the hepatopancreatic insulin, glucagon, leptin-AI and leptin-AII mRNA levels in goldfish during fasting and refeeding. Insulin was found to be effective in suppressing leptin-AI and leptin-AII transcript levels in goldfish liver via both in vivo intraperitoneal injection and in vitro cell incubation approaches. Only the insulin receptor, not the IGF-I receptor, was involved in insulin-inhibited leptin mRNA level. The suppression of leptin levels by insulin was caused by the activation of MKK3/6/p38MAPK and MEK1/2/Erk1/2 cascades. Insulin treatment could eliminate the stimulation of glucagon on leptin mRNA level. Our study describes the regulation and signal transduction mechanism of insulin on leptin mRNA levels in the goldfish liver, suggesting that the leptin function in fish is speculated to be not only an anorexigenic factor but also a metabolic mediator. This also supports the hypothesis that the poikilothermal fish use a passive survival strategy during the periods of food deprivation, which is mediated by the fish-specifically high leptin levels induced by the cooperation of insulin and glucagon.


Assuntos
Privação de Alimentos/fisiologia , Carpa Dourada/genética , Insulina/farmacologia , Leptina/genética , Fígado/metabolismo , Animais , Colforsina/farmacologia , Jejum , Comportamento Alimentar/efeitos dos fármacos , Glucagon/farmacologia , Hepatopâncreas/efeitos dos fármacos , Hepatopâncreas/metabolismo , Humanos , Injeções Intraperitoneais , Insulina/administração & dosagem , Fator de Crescimento Insulin-Like I/farmacologia , Fator de Crescimento Insulin-Like II/farmacologia , Leptina/metabolismo , Fígado/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores para Leptina/metabolismo , Transdução de Sinais/efeitos dos fármacos
8.
Artigo em Inglês | MEDLINE | ID: mdl-31176866

RESUMO

Cysteine oxygenase (CDO) is a mononuclear nonhemoglobin enzyme that catalyzes the production of taurine through the cysteine (Cys) pathway and plays a key role in the biosynthesis of taurine in mammals. However, the function of CDOs in bony fish remains poorly understood. In this study, we cloned CDO genes (CaCDO1 and CaCDO2) from Carassius auratus. The cDNA sequences of both CaCDO1 and CaCDO2 encoded putative proteins with 201 amino acids, which included structural features typical of the CDO protein family. Multiple sequence alignment and phylogenetic analysis showed that CaCDO1 and CaCDO2 shared high sequence identities and similarities with C. carpio homologs. Quantitative real-time polymerase chain reaction (qRT-PCR) results revealed that CaCDO1 and CaCDO2 were both broadly expressed in all selected tissues and developmental stages in C. auratus but had differing mRNA levels. In addition, compared to those of the taurine-free group, the in vivo mRNA expression levels of both CaCDO1 and CaCDO2 significantly decreased with increasing dietary taurine levels from 1.0 to 9.0 g/kg. Furthermore, in vitro taurine treatments showed similar inhibitory effects on the expression of CaCDO1 and CaCDO2 in the intestines of C. auratus. Our results also showed that the mRNA expression of CaCDO2 in the intestines was higher than that of CaCDO1 in response to in vivo and in vitro taurine supplementation. Overall, these data may provide new insights into the regulation of fish CDO expression and provide valuable knowledge for improving dietary formulas in aquaculture.


Assuntos
Cisteína Dioxigenase/genética , Cisteína Dioxigenase/metabolismo , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Carpa Dourada/genética , Carpa Dourada/metabolismo , Taurina/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA Complementar/genética , Regulação da Expressão Gênica no Desenvolvimento , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Carpa Dourada/crescimento & desenvolvimento , Isoenzimas/genética , Isoenzimas/metabolismo , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Taurina/farmacologia , Distribuição Tecidual
9.
Fish Shellfish Immunol ; 91: 78-86, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31039439

RESUMO

Type I interferons, as a class of multipotent cytokines, play a key role in host antiviral immune responses. In this study, a type I IFN coding gene of gibel carp, Carassius auratus gibelio, CagIFNa was cloned and sequenced. The full-length cDNA sequence of CagIFNa consists of 724 nucleotides that encode a predicted protein of 183 amino acids. CagIFNa has two highly conserved cysteine residues in the deduced protein, which is mostly conserved in the fish group I type I IFNs. CagIFNa was identified as a member of the IFNa subgroup of group I type I IFNs by phylogenetic analysis. CagIFNa transcripts were detected in all investigated tissues with higher levels in the liver, intestine, spleen and head kidney of gibel carp. Following injection with Cyprinid herpesvirus 2 (CyHV-2), CagIFNa gene expression was significantly inhibited in the spleen but delayed and then increased in head kidneys. Similarly, while CagIFNa expression was rapidly induced in gibel carp brain (GiCB) cells by poly I:C stimulation and its high induction level was delayed following CyHV-2 infection. CagIFNa overexpression in GiCB cells drastically reduced virus CPE and titer. Furthermore, several genes associated with type I IFN signaling pathway including IRF3, IRF7, IRF9, STAT1, Mx1 and PKR were induced in GiCB cells overexpressing CagIFNa upon CyHV-2 infection. These results show that CagIFNa plays a role in antiviral immune system in gibel carp.


Assuntos
Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Carpa Dourada/genética , Carpa Dourada/imunologia , Imunidade Inata/genética , Interferon Tipo I/genética , Interferon Tipo I/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Herpesviridae , Infecções por Herpesviridae/imunologia , Interferon Tipo I/química , Filogenia , Poli I-C/farmacologia , Distribuição Aleatória , Alinhamento de Sequência/veterinária
10.
J Immunol ; 202(8): 2407-2420, 2019 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-30850476

RESUMO

In mammals, tripartite motif (TRIM) proteins have emerged as pivotal players endowed with, directly, antiviral effects and, indirectly, modulatory capacity of the innate immune response. An unprecedented expansion of TRIM family has occurred in fish; however, the functional role of fish TRIM family members remains largely unknown. In this study, we identify a species-specific TRIM gene from crucian carp Carassius auratus, named FTRCA1, phylogenetically similar to the members of finTRIM, a subfamily of TRIM exclusively in teleost fish. FTRCA1 is induced by IFN and IFN stimuli as a typical IFN-stimulated gene. Overexpression of FTRCA1 negatively regulates IFN antiviral response by inhibition of IRF3 phosphorylation; consistently, knockdown of FTRCA1 results in enhanced levels of IRF3 phosphorylation and also IFN expression following poly(I:C) transfection. Whereas FTRCA1 is associated with several pivotal signaling molecules of RIG-I-like receptor pathway, its association with TBK1 results in autophage-lysosomal degradation of TBK1, thus abrogating the downstream IFN induction. Interestingly, FTRCA1 is phosphorylated by TBK1, but this phosphorylation is not required for downregulation of TBK1 protein. Transfection assays indicate that FTRCA1 is likely an E3 ligase with the requirement of RING finger domain, and deletion of N-terminal RING domain or mutation of seven conservative sites abolishes the negative regulatory function of FTRCA1. Collectively, these results illuminate a novel finTRIM-mediated innate immune modulatory pathway, thus providing insights into species-specific regulation of fish IFN response.


Assuntos
Autofagossomos/imunologia , Proteínas de Peixes/imunologia , Carpa Dourada/imunologia , Interferons/imunologia , Lisossomos/imunologia , Proteínas Serina-Treonina Quinases/imunologia , Proteólise , Proteínas com Motivo Tripartido/imunologia , Animais , Proteínas de Peixes/genética , Carpa Dourada/genética , Células HEK293 , Humanos , Interferons/genética , Lisossomos/genética , Proteínas Serina-Treonina Quinases/genética , Proteínas com Motivo Tripartido/genética
11.
Aquat Toxicol ; 209: 159-167, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30780113

RESUMO

The expression of estrogen receptors (ERs) and their roles in important cell processes such as apoptosis in the macrophages exposed to estrogen/xenoestrogen have remained a complex secret. This study focused on the expression of estrogen receptors (ERs) and the stimulation of apoptosis in the macrophages from the two sexes of goldfish (Carassius auratus) exposed to 17-ßestradiol (E2) and nonylphenol (NP) under in vivo and in vitro conditions. For the in vivo experiment, fish were exposed to NP (10-6 M and 10-7 M) and E2 (10-6 M) for 24 days. Then, the head kidney macrophages from the male and the female goldfish were isolated and assayed. For the in vitro experiments, the macrophages derived from the two sexes were cultured in L-15 medium and exposed to E2 (150 nM) and NP (10 nM and 150 nM) for 3 days. The results showed that the three isoforms of ERs (ERα, ERß1, ERß2) were expressed in the goldfish macrophages. After the exposure of macrophages to NP and E2, sex-specific increase of ERs expression and apoptosis were observed (P < 0.05). The expression of ERα after NP treatment showed the highest alteration, with the response being concentration-dependent. The most alteration of ERs expression were observed in the in vivo experiment. This study provides insight to understand how exposure of the goldfish macrophages to E2 and NP can up-regulate the transcript levels of estrogen receptor subtypes and stimulate apoptosis.


Assuntos
Receptor alfa de Estrogênio/metabolismo , Receptor beta de Estrogênio/metabolismo , Estrogênios/toxicidade , Carpa Dourada/metabolismo , Macrófagos/metabolismo , Modelos Biológicos , Fenóis/toxicidade , Animais , Apoptose/efeitos dos fármacos , Células Cultivadas , Receptor alfa de Estrogênio/genética , Receptor beta de Estrogênio/genética , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Carpa Dourada/genética , Macrófagos/efeitos dos fármacos , Masculino , Testes de Toxicidade , Poluentes Químicos da Água/toxicidade
12.
Mar Biotechnol (NY) ; 21(2): 139-149, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30426270

RESUMO

Autopolyploids are traditionally used to demonstrate multivalent pairing and unstable inheritance. However, the autotetraploid fish (4nRR) (RRRR, 4n = 200) derived from the distant hybridization of Carassius auratus red var. (RCC) (RR, 2n = 100) (♀) × Megalobrama amblycephala (BSB) (BB, 2n = 48) (♂) exhibits chromosome number (or ploidy) stability over consecutive generations (F1-F10). Comparative analysis based on somatic and gametic chromosomal loci [centromeric, 5S rDNA, and Ag-NORs (silver-stained nucleolar organizer regions)] revealed that a substantial loss of chromosomal loci during genome doubling increases the divergence between homologous chromosomes and that diploid-like chromosome pairing was restored during meiosis in the first generation of 4nRR lineages. In addition, a comparative analysis of genomes and transcriptomes from 4nRR (F1) and its diploid progenitor (RCC) exhibited significant genomic structure and gene expression changes. From these data, we suggest that genomes and genes diverge and that expression patterns change in the first generations following autotetraploidization, which are processes that might contribute to the stable inheritance and successful establishment of autotetraploid lineages.


Assuntos
Cyprinidae/genética , Carpa Dourada/genética , Hibridização Genética/genética , Tetraploidia , Animais , Cromossomos/genética , Feminino , Genoma , Hibridização in Situ Fluorescente , Masculino , Transcriptoma
13.
Fish Shellfish Immunol ; 84: 458-469, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30312666

RESUMO

Probiotic strains play an increasing role in the production of healthy animals used as a food source. Elucidating the mechanisms of action that allow probiotic-driven immunomodulation may facilitate different applications such as the prevention of infectious diseases in food organisms. This study elucidates the probiotic effects of Exiguobacterium acetylicum S01 on the growth, haematological profile, innate immune capacity, expression of cytokine genes, and resistance to diseases of Carassius auratus caused by Aeromonas hydrophila infection. Three fish groups were fed with the following diets containing different doses of E. acetylicum S01 (CFU g-1): basal diet 0 (BD, without probiotic), 2.5 × 107 (DI) and 2.7 × 109 (DII)-CFU g-1 for 4 weeks. After 4 weeks, the fish were injected intraperitoneally with A. hydrophila and the percentage of survival was recorded over 21 days of post-challenge. Results revealed that dietary supplementation of E. acetylicum S01 significantly (P < 0.05) enhanced the growth, haematological profile and cellular immune responses including respiratory burst, phagocytic activities and antimicrobial enzymes (myeloperoxidase and lysozyme) and total immunoglobulin levels were improved by probiotic feeding at both occasions. Comparatively, expression of c- and g-type lysozyme followed by pro- and anti-inflammatory cytokines (IL-1ß, IL-10 and TGFß) was up-regulated in kidney, head-kidney and spleen. Moreover, fish fed with diet DII had a significantly higher (P < 0.05) survival rate (73.2%) after challenging. The survival rate was only 33.2% of the BD group against A. hydrophila infection. Our results revealed that E. acetylicum S01 delivered probiotic in feed exerts its influence on growth performance and provides disease resistance by stimulating the immune system at the cellular and molecular levels in C. auratus.


Assuntos
Bacillales/química , Resistência à Doença/efeitos dos fármacos , Doenças dos Peixes/prevenção & controle , Regulação da Expressão Gênica/efeitos dos fármacos , Carpa Dourada/imunologia , Imunidade Inata/efeitos dos fármacos , Probióticos/farmacologia , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Citocinas/genética , Citocinas/metabolismo , Dieta/veterinária , Relação Dose-Resposta a Droga , Doenças dos Peixes/microbiologia , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Carpa Dourada/genética , Infecções por Bactérias Gram-Negativas/microbiologia , Infecções por Bactérias Gram-Negativas/prevenção & controle , Infecções por Bactérias Gram-Negativas/veterinária
14.
J Biochem ; 165(3): 209-218, 2019 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-30219851

RESUMO

The goldfish (Carassius auratus) is a domesticated cyprinid teleost closely related to the crucian carp. Goldfish domestication occurred in South China around 1,000 years ago. At least 180 variants and 70 genetically established strains are currently produced. These strains possess diverse phenotypes in body shape, colouration, scales, and fin, eye and hood morphology. These include biologically interesting phenotypes that have not been observed in mutants of zebrafish or medaka. In addition, goldfish strains have been maintained in a non-wild environment for several hundreds of generations, and certain goldfish strains have phenotypes similar to some human diseases. The recent progress in the assembly of the whole-genome sequence of goldfish provides strong tools for a genetic analysis of these phenotypes. The whole-genome duplication (WGD) event occurred in the goldfish genome 8-14 million years ago; this is one of the latest WGD in vertebrates. Goldfish are a useful model for studying genome evolution after the WGD event. This review focuses on the potential for goldfish as a model system in understanding the molecular basis of vertebrate development and evolution and human diseases.


Assuntos
Doença , Carpa Dourada/fisiologia , Modelos Animais , Animais , Doença/genética , Modelos Animais de Doenças , Carpa Dourada/genética , Carpa Dourada/crescimento & desenvolvimento , Humanos , Fenótipo
15.
Fish Shellfish Immunol ; 84: 1170-1179, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30366089

RESUMO

Stress response has negative effect on fish in aquaculture and research, which can be alleviated with anesthetic. To determine the optimal anesthetic, we investigated the physiological response of crucian carp (Carassius auratus) treated with three different anti-stress treatments: MS-222, eugenol and percussive stunning. Stress responses were evaluated by analyzing serum cortisol level and gene expression in blood. We determined the optimal concentrations of MS-222 (100 mg L-1) and eugenol (20 mg L-1) by dose selection. We found that the control group had significantly higher cortisol levels (172.78 ±â€¯19.95 ng mL-1) compared to the MS-222 treated group (46.85 ±â€¯3.22 ng mL-1), the eugenol treated group (72.78 ±â€¯9.07 ng mL-1), and the stunning treatment group (82.78 ±â€¯8.16 ng mL-1). Transcriptome analysis revealed 1572 differentially expressed genes (DEGs), including 155 DEGs related to the stress response, mainly involved in oxidative-stress response, heat shock proteins, and cold shock domain-containing protein. The heat shock protein genes were the primary DEGs in response to stress. RT-qPCR analysis confirmed differential expression of Hsps. We analyzed the function of the DEGs, which were enriched in genes involved in cellular response to stress and antigen processing and presentation. Combining the results from biochemical, transcriptome, and gene expression analysis, our data suggest that eugenol is more effective than MS-222 and percussive stunning in alleviating stress in crucian carp.


Assuntos
Aminobenzoatos/farmacologia , Anestesia/veterinária , Anestésicos/farmacologia , Carpas/fisiologia , Eugenol/farmacologia , Hidrocortisona/sangue , Anestesia/métodos , Animais , Carpas/genética , Relação Dose-Resposta a Droga , Expressão Gênica/efeitos dos fármacos , Perfilação da Expressão Gênica/veterinária , Carpa Dourada/genética , Carpa Dourada/fisiologia , Longevidade/efeitos dos fármacos , Estresse Fisiológico
16.
Fish Physiol Biochem ; 45(1): 427-437, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30361821

RESUMO

ß-Thymosins play critical roles in the regulation of many important physiological processes, but their function in teleost fishes remains poorly understood. In this study, the full-length cDNA coding for a thymosin ß (Tß) was cloned and identified in goldfish, Carassius auratus (gfTß). The gfTß cDNA consisted of 653 bp with an open reading frame of 135 bp that encodes a 44 amino acid polypeptide. Sequence analysis revealed one thymosin domain and a highly conserved actin-binding motif (18LKKTET23). Expression of gfTß transcript was detected ubiquitously in all tissues examined, with relatively higher levels in the brain, intestine, spleen, gill, skin, kidney, and testis. Cadmium and H2O2 exposure induced increases in gfTß transcript levels in the liver and spleen. Moreover, gfTß transcription was upregulated in response to LPS challenge in the spleen while Poly I:C treatment did not affect gfTß expression. In vivo injection of recombinant gfTß generated from an Escherichia coli system induced expression of T lymphocyte-related genes (RAG1 and CD8α). These results suggest that gfTß may be involved in the immune response of teleost fishes via modulation of T lymphocyte development.


Assuntos
Clonagem Molecular , Regulação da Expressão Gênica/fisiologia , Carpa Dourada/metabolismo , Timosina/metabolismo , Animais , Carpa Dourada/genética , Carpa Dourada/imunologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Timosina/genética
17.
Fish Shellfish Immunol ; 86: 913-921, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30550991

RESUMO

The present study was conducted to evaluate dietary inosine 5'-monophosphate (5'-IMP) on growth, immune genes expression and disease resistance against Aeromonas hydrophila in juvenile gibel carp (Carassius auratus gibelio var. CAS Ⅲ) (initial body weight: 7.48 g). Six diets were formulated containing exogenous 5'-IMP at three gradient levels (0, 0.1% and 0.2%) in the high dietary fishmeal group (15% fishmeal: D1, D2, D3) and in the high dietary soybean meal group (33% soybean meal: D4, D5, D6). Each diet was randomly allotted to triplicate tanks in a recirculating system. After the feeding trial, fish were exposed to Aeromonas hydrophila challenge. Hematological and immunological responses were analyzed before and after challenge. The results indicated that feeding rate in all 5'-IMP supplemented treatments (D2, D3, D5 and D6) and daily growth coefficient in D5 and D6 were reduced compared with those of respective control treatments (D1 and D4) without 5'-IMP addition (P < 0.05). The cumulative survival rates were numerically improved by dietary 5'-IMP supplementation (P > 0.05). Compared with the respective control treatment, in the high fishmeal group, plasma SOD and MPO were significantly elevated in D3 at the end of feeding trial (P < 0.05), plasma SOD and lysozyme were significantly increased in D3 after bacterial challenge (P < 0.05); in high soybean meal group, plasma lysozyme activity was significantly elevated in D5 post bacterial challenge (P < 0.05). Most of the expression of immune related genes (intelectin, major histocompatibility complex class II ß (MHC II ß), Complement 3 (C3), Complement component C7-1 (ccC7), lysozyme C, Interleukin 1ß (IL-1ß), Tumor necrosis factor α1 (TNF-α1), Transforming growth factor-beta (TGF-ß) and Interleukin 8 (IL-8)) in spleen, kidney and liver of the fish were significantly affected by supplementation of 5'-IMP at the end of feeding trial and post bacterial challenge. Additionally, adding 5'-IMP in high soybean meal diets exerted further effects of promoting immunity than counterparts in high fishmeal diets. Considering enhanced disease resistance, the immunopotentiation of 5'-IMP was manifested when the addition level was 0.1% in high soybean meal diets and 0.2% in high fishmeal diets.


Assuntos
Dieta/veterinária , Regulação da Expressão Gênica/imunologia , Carpa Dourada/genética , Carpa Dourada/imunologia , Inosina Monofosfato/metabolismo , Aeromonas hydrophila/fisiologia , Ração Animal/análise , Animais , Dieta/classificação , Suplementos Nutricionais/análise , Resistência à Doença/efeitos dos fármacos , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Carpa Dourada/crescimento & desenvolvimento , Infecções por Bactérias Gram-Negativas/imunologia , Infecções por Bactérias Gram-Negativas/veterinária , Inosina Monofosfato/administração & dosagem
18.
J Exp Zool B Mol Dev Evol ; 330(6-7): 372-383, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30387925

RESUMO

The twin tail of ornamental goldfish is known to be caused by a nonsense mutation in one chordin paralogue gene. Our previous molecular studies in goldfish revealed that the ancestral chordin gene was duplicated, creating the chdA and chdB genes, and the subsequent introduction of a stop codon allele in the chdA gene ( chdA E127X ) caused the twin-tail morphology. The chdA E127X allele was positively selected by breeders, and the allele was genetically fixed in the ornamental twin-tail goldfish population. However, little is known about the evolutionary history of the chdB paralogue, begging the question: are there the functionally distinct alleles at the chdB locus, and if so, how did they evolve? To address these questions, we conducted molecular sequencing of the chdB gene from five different goldfish strains and discovered two alleles at the chdB gene locus; the two alleles are designated chdB 1 and chdB 2 . The chdB 1 allele is the major allele and was found in all investigated goldfish strains, whereas the chdB 2 allele is minor, having only been found in one twin-tail strain. Genetic analyses further suggested that these two alleles are functionally different with regard to survivability ( chdB 1 > chdB 2 ). These results led us to presume that in contrast to the chdA locus, the chdB locus has tended to be eliminated from the population. We also discuss how the chdB 2 allele was retained in the goldfish population, despite its disadvantageous function. This study provides empirical evidence of the long-term retention of a disadvantageous allele under domesticated conditions.


Assuntos
Nadadeiras de Animais , Glicoproteínas/genética , Carpa Dourada/anatomia & histologia , Carpa Dourada/genética , Peptídeos e Proteínas de Sinalização Intercelular/genética , Alelos , Animais , Evolução Molecular , Morfogênese/genética , Análise de Sequência de DNA
19.
Sci China Life Sci ; 61(11): 1407-1419, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30178265

RESUMO

Focusing on adaptation of aquatic organisms, especially fish, can help elucidate complex dynamics in freshwater ecology. The differences in genetic and epigenetic regulation between diploid and triploid Carassius auratus affect survival under eutrophication. To identify the underlying mechanisms that lead to better adaption of triploids than diploids, we compared mRNA and microRNA (miRNA) expressions in liver tissue of diploid and triploid individuals obtained from the Dongting lake water system in central China. Differential expression analysis revealed that 566 transcripts were significantly up-regulated, whereas 758 were down-regulated in triploids; of these differentially expressed transcripts, 33 transcripts including cacna1d, nfkb2, hspa1 and fgfr4 were involved in the MAPK signaling pathway, and eight transcripts were determined to be regulated by seven miRNAs. Additionally, four of 25 differential expressed (DE) transcripts (mhc1, irf7, nfkb2 and pik3c) involving the viral carcinogenesis pathway were regulated by four miRNAs. Furthermore, genetic polymorphisms analysis showed that more heterozygous mutations were detected in triploids than diploids. The dN/dS results revealed that 21 genes were under positive selection (dN/dS>1) in C. auratus complex. We hypothesize that these changes related to genetic and epigenetic regulation may be caused by abiotic stresses, and facilitate adaptation to environmental changes.


Assuntos
Adaptação Fisiológica/genética , Mudança Climática , Carpa Dourada/fisiologia , Animais , China , Diploide , Monitoramento Ambiental , Epigênese Genética , Perfilação da Expressão Gênica , Carpa Dourada/genética , Lagos , MicroRNAs/genética , Polimorfismo Genético , RNA Mensageiro/genética , Seleção Genética , Triploidia
20.
J Fish Biol ; 93(6): 1082-1089, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30260011

RESUMO

Autotetraploid fish (4n = 200, abbreviated as 4nRR), which reach sexual maturity at 1 year of age, were derived from the whole genome duplication of red crucian carp Carassius auratus red var. (RCC; 2n = 100) and possess four sets of chromosomes from RCC. The histological features of the gonads showed that the RCC and 4nRR both possessed normal gonadal structure and could arrive at maturation. To understand the expression characteristics of genes related to reproductive development in the autotetraploid fish, we analysed the nucleotide sequence and expression characteristics of the gnrh2, gthb and gthr genes, which are the pivotal genes of the hypothalamic-pituitary-gonadal (HPG) axis. We found that the gnrh2, gthb and gthr genes in 4nRR share remarkable homology with RCC, but there were obvious differences in expression levels between 4nRR and RCC. These results demonstrate that autotetraploidization can lead to gene expression changes. This study provides insights into the molecular mechanism underlying the reproductive development of autotetraploid fish and is expected to be of great significance for subsequent research on polyploidization.


Assuntos
Expressão Gênica , Carpa Dourada/genética , Reprodução , Animais , Duplicação Cromossômica , Regulação da Expressão Gênica no Desenvolvimento , Carpa Dourada/anatomia & histologia , Carpa Dourada/fisiologia , Gônadas/anatomia & histologia , Gônadas/metabolismo , Gônadas/fisiologia , Sistema Hipotálamo-Hipofisário , Maturidade Sexual/genética , Tetraploidia
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