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1.
PLoS Pathog ; 16(7): e1008668, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32639977

RESUMO

Nervous necrosis virus (NNV) can infect many species of fish and causes serious acute or persistent infection. However, its pathogenic mechanism is still far from clear. Specific cellular surface receptors are crucial determinants of the species tropism of a virus and its pathogenesis. Here, the heat shock protein 90ab1 of marine model fish species marine medaka (MmHSP90ab1) was identified as a novel receptor of red-spotted grouper NNV (RGNNV). MmHSP90ab1 interacted directly with RGNNV capsid protein (CP). Specifically, MmHSP90ab1 bound to the linker region (LR) of CP through its NM domain. Inhibition of MmHSP90ab1 by HSP90-specific inhibitors or MmHSP90ab1 siRNA caused significant inhibition of viral binding and entry, whereas its overexpression led to the opposite effect. The binding of RGNNV to cultured marine medaka hMMES1 cells was inhibited by blocking cell surface-localized MmHSP90ab1 with anti-HSP90ß antibodies or pretreating virus with recombinant MmHSP90ab1 or MmHSP90ab1-NM protein, indicating MmHSP90ab1 was an attachment receptor for RGNNV. Furthermore, we found that MmHSP90ab1 formed a complex with CP and marine medaka heat shock cognate 70, a known NNV receptor. Exogenous expression of MmHSP90ab1 independently facilitated the internalization of RGNNV into RGNNV impenetrable cells (HEK293T), which was blocked by chlorpromazine, an inhibitor of clathrin-dependent endocytosis. Further study revealed that MmHSP90ab1 interacted with the marine medaka clathrin heavy chain. Collectively, these data suggest that MmHSP90ab1 is a functional part of the RGNNV receptor complex and involved in the internalization of RGNNV via the clathrin endocytosis pathway.


Assuntos
Doenças dos Peixes/metabolismo , Proteínas de Peixes/metabolismo , Proteínas de Choque Térmico/metabolismo , Infecções por Vírus de RNA/veterinária , Receptores Virais/metabolismo , Animais , Clatrina/metabolismo , Endocitose , Peixes , Nodaviridae/metabolismo , Oryzias/virologia , Internalização do Vírus
2.
Proc Natl Acad Sci U S A ; 117(32): 19321-19327, 2020 08 11.
Artigo em Inglês | MEDLINE | ID: mdl-32719137

RESUMO

Phenotypic plasticity, the ability of a single genotype to produce multiple phenotypes under different environmental conditions, is critical for the origins and maintenance of biodiversity; however, the genetic mechanisms underlying plasticity as well as how variation in those mechanisms can drive evolutionary change remain poorly understood. Here, we examine the cichlid feeding apparatus, an icon of both prodigious evolutionary divergence and adaptive phenotypic plasticity. We first provide a tissue-level mechanism for plasticity in craniofacial shape by measuring rates of bone deposition within functionally salient elements of the feeding apparatus in fishes forced to employ alternate foraging modes. We show that levels and patterns of phenotypic plasticity are distinct among closely related cichlid species, underscoring the evolutionary potential of this trait. Next, we demonstrate that hedgehog (Hh) signaling, which has been implicated in the evolutionary divergence of cichlid feeding architecture, is associated with environmentally induced rates of bone deposition. Finally, to demonstrate that Hh levels are the cause of the plastic response and not simply the consequence of producing more bone, we use transgenic zebrafish in which Hh levels could be experimentally manipulated under different foraging conditions. Notably, we find that the ability to modulate bone deposition rates in different environments is dampened when Hh levels are reduced, whereas the sensitivity of bone deposition to different mechanical demands increases with elevated Hh levels. These data advance a mechanistic understanding of phenotypic plasticity in the teleost feeding apparatus and in doing so contribute key insights into the origins of adaptive morphological radiations.


Assuntos
Ciclídeos/metabolismo , Proteínas de Peixes/metabolismo , Proteínas Hedgehog/metabolismo , Crânio/crescimento & desenvolvimento , Adaptação Fisiológica , Animais , Ciclídeos/genética , Ciclídeos/crescimento & desenvolvimento , Proteínas de Peixes/genética , Proteínas Hedgehog/genética , Transdução de Sinais , Crânio/metabolismo
3.
Proc Natl Acad Sci U S A ; 117(32): 19276-19286, 2020 08 11.
Artigo em Inglês | MEDLINE | ID: mdl-32719141

RESUMO

Bone homeostasis requires continuous remodeling of bone matrix to maintain structural integrity. This involves extensive communication between bone-forming osteoblasts and bone-resorbing osteoclasts to orchestrate balanced progenitor cell recruitment and activation. Only a few mediators controlling progenitor activation are known to date and have been targeted for intervention of bone disorders such as osteoporosis. To identify druggable pathways, we generated a medaka (Oryzias latipes) osteoporosis model, where inducible expression of receptor-activator of nuclear factor kappa-Β ligand (Rankl) leads to ectopic formation of osteoclasts and excessive bone resorption, which can be assessed by live imaging. Here we show that upon Rankl induction, osteoblast progenitors up-regulate expression of the chemokine ligand Cxcl9l. Ectopic expression of Cxcl9l recruits mpeg1-positive macrophages to bone matrix and triggers their differentiation into osteoclasts. We also demonstrate that the chemokine receptor Cxcr3.2 is expressed in a distinct subset of macrophages in the aorta-gonad-mesonephros (AGM). Live imaging revealed that upon Rankl induction, Cxcr3.2-positive macrophages get activated, migrate to bone matrix, and differentiate into osteoclasts. Importantly, mutations in cxcr3.2 prevent macrophage recruitment and osteoclast differentiation. Furthermore, Cxcr3.2 inhibition by the chemical antagonists AMG487 and NBI-74330 also reduced osteoclast recruitment and protected bone integrity against osteoporotic insult. Our data identify a mechanism for progenitor recruitment to bone resorption sites and Cxcl9l and Cxcr3.2 as potential druggable regulators of bone homeostasis and osteoporosis.


Assuntos
Matriz Óssea/metabolismo , Quimiocina CXCL9/metabolismo , Proteínas de Peixes/metabolismo , Oryzias/metabolismo , Osteoclastos/metabolismo , Osteoporose/metabolismo , Receptores CXCR3/metabolismo , Células-Tronco/metabolismo , Animais , Matriz Óssea/crescimento & desenvolvimento , Diferenciação Celular , Quimiocina CXCL9/genética , Modelos Animais de Doenças , Proteínas de Peixes/genética , Humanos , Macrófagos/metabolismo , Oryzias/genética , Oryzias/crescimento & desenvolvimento , Osteoblastos/citologia , Osteoblastos/metabolismo , Osteoclastos/citologia , Osteoporose/genética , Osteoporose/fisiopatologia , Ligação Proteica , Receptores CXCR3/genética , Células-Tronco/citologia
4.
PLoS One ; 15(7): e0236601, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32730353

RESUMO

Omega-3 polyunsaturated fatty acids (n-3 PUFAs), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), exhibit antibacterial and anti-inflammatory activities. Furthermore, diets rich in n-3 PUFAs are known to improve disease resistance and limit pathogen infection in commercial aquaculture fishes. In this study, we examined the effects of transgenic overexpression of n-3 PUFA biosynthesis genes on the physiological response to bacterial infection in tilapia. We first established tilapia strains with single or dual expression of salmon delta-5 desaturase and/or delta-6 desaturase and then challenged the fish with Vibrio vulnificus infection. Interestingly, our data suggest that n-3 PUFA-mediated alterations in gut microbiota may be important in determining disease outcome via effects on immune response of the host. Both liver- and muscle-specific single and dual expression of delta-5 desaturase and delta-6 desaturase resulted in higher n-3 PUFA content in transgenic fish fed with a LO basal diet. The enrichment of n-3 PUFAs in dual-transgenic fish is likely responsible for their improved survival rate and comparatively reduced expression of inflammation- and immune-associated genes after V. vulnificus infection. Gut microbiome analysis further revealed that dual-transgenic tilapia had high gut microbiota diversity, with low levels of inflammation-associated microbiota (i.e., Prevotellaceae). Thus, our findings indicate that dual expression of transgenic delta-5 and delta-6 desaturase in tilapia enhances disease resistance, an effect that is associated with increased levels of n-3 PUFAs and altered gut microbiota composition.


Assuntos
Resistência à Doença , Ácidos Graxos Dessaturases/metabolismo , Proteínas de Peixes/metabolismo , Microbioma Gastrointestinal , Linoleoil-CoA Desaturase/metabolismo , Tilápia/microbiologia , Vibrio vulnificus/patogenicidade , Animais , Animais Geneticamente Modificados/genética , Animais Geneticamente Modificados/microbiologia , Dieta/veterinária , Análise Discriminante , Resistência à Doença/genética , Ácidos Docosa-Hexaenoicos/metabolismo , Ácidos Graxos Dessaturases/genética , Ácidos Graxos Ômega-3/metabolismo , Doenças dos Peixes/microbiologia , Doenças dos Peixes/patologia , Proteínas de Peixes/genética , Expressão Gênica , Análise dos Mínimos Quadrados , Linoleoil-CoA Desaturase/genética , Tilápia/genética , Vibrioses/patologia , Vibrioses/veterinária
5.
Food Funct ; 11(6): 5565-5572, 2020 Jun 24.
Artigo em Inglês | MEDLINE | ID: mdl-32520031

RESUMO

To date, no specific drug has been discovered for the treatment of COVID-19 and hence, people are in a state of anxiety. Thus, there is an urgent need to search for various possible strategies including nutritional supplementation. In this study, we have tried to provide a reference for protein supplementation. Specifically, 20 marine fish proteins were subjected to in silico hydrolysis by gastrointestinal enzymes, and a large number of active peptides were generated. Then, the binding abilities of these peptides to SARS-CoV-2 main protease and monoamine oxidase A were assessed. The results showed that NADH dehydrogenase could be a good protein source in generating potent binders to the two enzymes, followed by cytochrome b. In addition, some high-affinity oligopeptides (VIQY, ICIY, PISQF, VISAW, AIPAW, and PVSQF) were identified as dual binders to the two enzymes. In summary, the supplementation of some fish proteins can be helpful for COVID-19 patients; the identified oligopeptides can be used as the lead compounds to design potential inhibitors against COVID-19 and anxiety.


Assuntos
Antivirais/metabolismo , Betacoronavirus/metabolismo , Infecções por Coronavirus/virologia , Suplementos Nutricionais , Proteínas de Peixes/metabolismo , Monoaminoxidase/metabolismo , Pneumonia Viral/virologia , Animais , Antivirais/química , Antivirais/uso terapêutico , Organismos Aquáticos , Betacoronavirus/enzimologia , Infecções por Coronavirus/tratamento farmacológico , Decapodiformes/metabolismo , Proteínas de Peixes/química , Proteínas de Peixes/uso terapêutico , Peixes/metabolismo , Modelos Moleculares , Simulação de Acoplamento Molecular , Inibidores da Monoaminoxidase , Pandemias , Perciformes/metabolismo , Pneumonia Viral/tratamento farmacológico , Ligação Proteica , Conformação Proteica , Salmão/metabolismo , Atum/metabolismo
6.
PLoS One ; 15(5): e0233322, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32469895

RESUMO

The importance of dietary lipids in male reproduction are not as well understood as in females, in which dietary lipids, such as phospholipids (PL) and associated fatty acids (FA), are important structural components of the eggs and provide energy for their offspring. In mammals, lipids are suggested to be important for spermatogenesis and to structural components of the spermatozoa that could improve fertilization rates. New knowledge of how lipids affect sexual maturation in male Atlantic salmon (Salmo salar), an important global aquaculture species, could provide tools to delay maturation and/or improve reproductive success. Therefore, changes in testicular composition of lipids and gene transcripts associated with spermatogenesis and lipid metabolism were studied in sexually maturing male salmon compared to immature males and females. An increase in total testis content of FA and PL, and a shift to higher PL composition was observed in maturing males, concomitant with increases in mRNA levels for genes involved in spermatogenesis, FA uptake and synthesis, and production of long chain-polyunsaturated fatty acids (LC-PUFA) and PL. A particularly interesting finding was elevated testis expression of acyl-CoA synthetase 4 (acsl4), and acyl-CoA thioesterase 2 (acot2), critical enzymes that regulate intra-mitochondrial levels of 20:4n-6 FA (arachidonic acid), which have been associated with improved cholesterol transport during steroidogenesis. This suggested that FA may have direct effects on sex steroid production in salmon. Furthermore, we observed increased testis expression of genes for endogenous synthesis of 16:0 and elongation/desaturation to 22:6n-3 (docosahexaenoic acid) in sexually maturing males relative to immature fish. Both of these FA are important structural components of the PL, phosphatidylcholine (PC), and were elevated concomitant with increases in the content of phosphatidic acid, an important precursor for PC, in maturing males compared to immature fish. Overall, this study suggests that, similar to mammals, lipids are important to spermatogenesis and serve as structural components during testicular growth and maturation in Atlantic salmon.


Assuntos
Ácidos Graxos Insaturados/metabolismo , Proteínas de Peixes/metabolismo , Regulação da Expressão Gênica , Fosfolipídeos/metabolismo , Salmo salar/metabolismo , Maturidade Sexual , Testículo/metabolismo , Animais , Ácidos Graxos Dessaturases/genética , Feminino , Proteínas de Peixes/genética , Perfilação da Expressão Gênica , Masculino , Salmo salar/genética , Testículo/citologia
7.
Food Chem ; 327: 126777, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32446027

RESUMO

The micro-flowing water system can improve the flesh quality of freshwater fish using the traditional pond farming method. However, the mechanism of this phenomenon has not yet been explored. This study intends to examine the changes of metabolites in freshwater fish after treatment with the micro-flowing purification system (MFPS). The UPLC-QTOF/MS based metabolomics method was utilized to screen the metabolites and predict the major possible metabolic pathways after MFPS treatment. There were 377 types of metabolites identified in the fish muscle, of which 54-71 represented significant different metabolites identified during different stages of MFPS treatments. The main mechanism of MFPS treatment in improving the quality of grass carp fish muscle was investigated, and the MFPS treatment was shown to improve the flesh quality and the flavor of grass carp fish muscle. This study could provide the theoretical basis for improving the quality of aquatic products.


Assuntos
Carpas/metabolismo , Metabolômica , Animais , Cromatografia Líquida de Alta Pressão , Proteínas de Peixes/metabolismo , Água Doce , Espectrometria de Massas , Músculo Esquelético/metabolismo , Purificação da Água
8.
J Environ Sci Health B ; 55(8): 687-693, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32432968

RESUMO

The objective of this research was to investigate the potential damage caused by the residual concentrations of the insecticides Regent® WS 800 and Curbix® SC 200, containing fipronil and ethiprole, respectively as active ingredients, on the liver of Oreochromis niloticus. The analyses of HSP70 shock protein labelling and cell death process by TUNEL method were performed in order to measure the effects of the exposure of cell repair system of fish to both insecticides. Statistical analyses showed no significant molecular damage to the hepatic tissue of animals. Nevertheless, variations in HSP70 and DNA fragmentation levels, endpoint of cell repair system response and cellular death, respectively, were observed in several groups. These results indicate that the cell repair machinery was efficient when in contact with residual concentrations of insecticides. However, the DNA fragmentation detected by the TUNEL method suggests that even in face of the cytoprotective action of the HSP70 protein, there are damages that become irreparable. To finish, it is worth mentioning that given the results obtained from residual concentrations, use in the field should be with caution.


Assuntos
Ciclídeos/fisiologia , Proteínas de Choque Térmico HSP70/metabolismo , Inseticidas/toxicidade , Pirazóis/toxicidade , Animais , Morte Celular/efeitos dos fármacos , Biomarcadores Ambientais/efeitos dos fármacos , Proteínas de Peixes/metabolismo , Fígado/efeitos dos fármacos , Resíduos de Praguicidas/toxicidade , Poluentes Químicos da Água/toxicidade
9.
Proc Natl Acad Sci U S A ; 117(22): 12174-12181, 2020 06 02.
Artigo em Inglês | MEDLINE | ID: mdl-32409601

RESUMO

Germ cells have the ability to differentiate into eggs and sperm and must determine their sexual fate. In vertebrates, the mechanism of commitment to oogenesis following the sexual fate decision in germ cells remains unknown. Forkhead-box protein L3 (foxl3) is a switch gene involved in the germline sexual fate decision in the teleost fish medaka (Oryzias latipes). Here, we show that foxl3 organizes two independent pathways of oogenesis regulated by REC8 meiotic recombination protein a (rec8a), a cohesin component, and F-box protein (FBP) 47 (fbxo47), a subunit of E3 ubiquitin ligase. In mutants of either gene, germ cells failed to undergo oogenesis but developed normally into sperm in testes. Disruption of rec8a resulted in arrest at a meiotic pachytenelike stage specifically in females, revealing a sexual difference in meiotic progression. Analyses of fbxo47 mutants showed that this gene regulates transcription factors that facilitate folliculogenesis: LIM homeobox 8 (lhx8b), factor in the germline α (figla), and newborn ovary homeobox (nobox). Interestingly, we found that the fbxo47 pathway ensures that germ cells do not deviate from an oogenic pathway until they reach diplotene stage. The mutant phenotypes together with the timing of their expression imply that germline feminization is established during early meiotic prophase I.


Assuntos
Proteínas de Peixes/metabolismo , Células Germinativas/citologia , Gônadas/embriologia , Oogênese , Oryzias/crescimento & desenvolvimento , Folículo Ovariano/fisiologia , Espermatozoides/fisiologia , Animais , Feminino , Proteínas de Peixes/genética , Regulação da Expressão Gênica no Desenvolvimento , Células Germinativas/fisiologia , Gônadas/metabolismo , Masculino , Meiose , Folículo Ovariano/citologia , Processos de Determinação Sexual , Espermatogênese , Espermatozoides/citologia
10.
Food Chem ; 327: 127079, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32446028

RESUMO

Two water-soluble red color-related proteins with the molecular masses of 24 and 73 kDa were purified from the shell of Procambarus clarkii. Initial color changes of these two proteins were detected at 30 °C and the large amount of red precipitate were obtained at 80 °C. PAGE analysis showed that the 24 kDa protein was the monomer, while the 73 kDa protein was the trimer. Identification revealed that these two proteins belonged to the hemocyanin subunit 2 family. With respect to the amino acid sequence similarity, the red color-related proteins shared the highest sequence identity with the hemocyanin derived from giant freshwater prawn (Macrobrachium rosenbergii). The phylogenetic tree analysis also clearly supported this finding. The shell-derived red color-related proteins show potential use as the edible thermal-sensitive indicator in food processing field.


Assuntos
Astacoidea/química , Proteínas de Peixes/química , Sequência de Aminoácidos , Animais , Astacoidea/genética , Astacoidea/metabolismo , Cor , Biologia Computacional , Proteínas de Peixes/genética , Proteínas de Peixes/isolamento & purificação , Proteínas de Peixes/metabolismo , Hemocianinas/química , Hemocianinas/metabolismo , Filogenia
11.
BMC Bioinformatics ; 21(1): 174, 2020 May 04.
Artigo em Inglês | MEDLINE | ID: mdl-32366294

RESUMO

BACKGROUND: Transcriptome analysis by next-generation sequencing has become a popular technique in recent years. This approach is quite suitable for non-model organism study, as de novo assembly is independent of prior genomic sequences of organisms. De novo sequencing has benefited many studies on commercially important fish species. However, to understand the functions of these assembled sequences, they still need to be annotated with existing sequence databases. By combining Basic Local Alignment Search Tool (BLAST) and Gene Ontology analysis, we were able to identify homologous sequences of assembled sequences and describe their characteristics using pre-defined tags for each gene, though the above conventional annotation results obtained for non-model assembled sequences was still associated with a lack of pre-defined tags and poorly documented records in the database. RESULTS: We introduced Blast2Fish, a novel approach for performing functional enrichment analysis on non-model teleost fish transcriptome data. The Blast2Fish pipeline was designed to be a reference-based enrichment method. Instead of annotating the BLAST single top hit by a pre-defined gene-to-tag database, we included 500 hits to search related PubMed articles and parse biological terms. These descriptive terms were then sorted and recorded as annotations for the query. The results showed that Blast2Fish was capable of providing meaningful annotations on immunology topics for non-model fish transcriptome analysis. CONCLUSION: Blast2Fish provides a novel approach for annotating sequences of non-model fish. The reference-based strategy allows annotation to be performed without pre-defined tags for each gene. This method strongly benefits non-model teleost fish studies for gene functional enrichment analysis.


Assuntos
Biologia Computacional/métodos , Proteínas de Peixes/genética , Peixes/genética , Anotação de Sequência Molecular/métodos , Animais , Bases de Dados de Ácidos Nucleicos , Proteínas de Peixes/química , Proteínas de Peixes/metabolismo , Peixes/metabolismo , Perfilação da Expressão Gênica , Genômica , Sequenciamento de Nucleotídeos em Larga Escala , Internet , Software , Transcriptoma
12.
Zoolog Sci ; 37(2): 177-192, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32282149

RESUMO

There are few model fish that are both edible and suitable for use in the laboratory. The Japanese loach (Misgurnus anguillicaudatus) is a traditional food in Japan, but is highly neglected despite its great nutritional value. To understand its circadian system and photic input pathway for synchronization of physiological activities to environmental light-dark cycles, we measured locomotor activity under light-dark and constant dark (DD) conditions. Locomotor activity was found to be higher in the nighttime than daytime, and its rhythmicity was weakened under DD conditions. The nocturnal activity of the Japanese loach is mainly controlled by environmental light, rather than the circadian clock. We explored the circadian regulation and light-responsiveness of clock gene expression in the eyes of loaches. The daily expression profiles of its mRNA revealed that most of the examined Cry and Per genes were likely regulated by internal circadian and/or environmental light signals. Among the Opsin genes transcribed in the eye, we detected the retinal photopigment porphyropsin at the protein level, which was lower than in mice. This property of loach eyes prompted us to analyze the locomotor activities of eye-enucleated fish. As a result, they still showed nocturnal circadian activity. Thus, it is likely that extraocular photoreceptive tissue(s) also contribute to the photic input pathway, although loach eyes are a circadian photosensitive tissue. This suggests that the loach mainly uses not its vision but other stimuli, such as mechanical or chemical stimuli, detected by barbels, to coordinate its nocturnal behavior.


Assuntos
Relógios Circadianos/genética , Cipriniformes/genética , Animais , Relógios Circadianos/fisiologia , Ritmo Circadiano/fisiologia , Cipriniformes/fisiologia , Olho/química , Olho/metabolismo , Feminino , Proteínas de Peixes/metabolismo , Regulação da Expressão Gênica , Luz , Locomoção/fisiologia , Masculino , Fenômenos Fisiológicos Oculares , RNA Mensageiro
13.
Gene ; 745: 144651, 2020 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-32259633

RESUMO

Sexual differentiation and ovotestis development are closely associated with cortisol levels, the principal indicator of stress, via the glucocorticoid receptor (GR) in teleosts. Thus, GR is regarded as a mediator to expound the relationship between social stress and gonad development. In the present study, two gr genes (gr1 and gr2) were cloned and analyzed from a protandrous hermaphroditic teleost, the yellowtail clownfish (Amphiprion clarkii). GR1 was found to display a conserved nine-amino-acid insert, WRARQNTDG, between two zinc finger domains. The phylogenetic tree of GR showed that yellowtail clownfish GR1 and GR2 are clustered to teleost GR1 and teleost GR2 separately, and differ from tetrapod GR. The result of real-time PCR revealed that high-level gr1 was mainly distributed in the cerebellum, hypothalamus and heart. The gr2 gene was abundant in the pituitary and liver of females and nonbreeders, while gr2 was mainly detected in the medulla oblongata and middle kidney of males. Moreover, GRs can be expressed in cultured eukaryotic cells and functionally interact with dexamethasone (exogenous glucocorticoid), thereby triggering downstream signaling pathways of different potentials. GR1 and GR2 can be activated by 10 nM dexamethasone treatment in HEK-293T cells. Notably, real-time PCR analysis among three social status groups demonstrated that gr2 expression was the highest in the hypothalamus of nonbreeders, but gr1 was no difference. We speculate that social stress would increase the expression of gr2 gene expression in the hypothalamus to inhibit sexual development. These data provide evidence of social stress involving reproductive regulation, which may help to elucidate the underlying mechanism of sex differentiation and change.


Assuntos
Proteínas de Peixes/genética , Regulação da Expressão Gênica no Desenvolvimento , Perciformes/genética , Receptores de Glucocorticoides/genética , Diferenciação Sexual/genética , Animais , Dexametasona/farmacologia , Feminino , Proteínas de Peixes/metabolismo , Células HEK293 , Humanos , Masculino , Família Multigênica , Perciformes/metabolismo , Filogenia , Receptores de Glucocorticoides/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Reprodução/genética , Transdução de Sinais/efeitos dos fármacos , Estresse Psicológico/genética , Dedos de Zinco/genética
14.
Vet Res ; 51(1): 45, 2020 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-32197655

RESUMO

Yersinia ruckeri is a facultative intracellular enterobacterium mostly known as the causative agent of enteric redmouth disease in salmonid fish. In the present study, we applied RNA inhibition to silence twenty pre-selected genes on the genome of a fish cell line (CHSE-214) followed by a gentamicin assay to quantify the effect of silencing on the cells' susceptibility to infection and found that silencing of 18 out of 20 genes significantly reduced the number of Y. ruckeri recovered. These findings improve our understanding of the infection process by Y. ruckeri and of the interactions between this bacterial pathogen and host cells.


Assuntos
Doenças dos Peixes/genética , Proteínas de Peixes/genética , Inativação Gênica , RNA Interferente Pequeno/genética , Yersiniose/veterinária , Animais , Linhagem Celular , Doenças dos Peixes/microbiologia , Proteínas de Peixes/metabolismo , RNA Interferente Pequeno/metabolismo , Yersiniose/genética , Yersiniose/microbiologia , Yersinia ruckeri/fisiologia
15.
Res Vet Sci ; 130: 212-221, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32203766

RESUMO

A trial was operated to assess the potential of using Lactobacillus plantarum L-137 (L-137) and/or ß-glucan (BG) in improving the resistance of Nile tilapia against Aeromonas hydrophila. Control diet and 3 diets supplemented with L-137, BG or L-137 + BG were prepared. Final body weight, specific growth rate, superoxide dismutase, and catalase showed considerably (P < .05) increased values in L-137 or L-137/BG groups, while glutathione peroxidase increased significantly (P < .05) only in L-137/BG group. Fish fed L-137 and/or BG diets showed that feed conversion ratio and malonaldehyde levels were significantly decreased (P < .05). Also, both L-137 and BG helped Nile tilapia to have high phagocytosis activity and relative expression of tumor necrosis factor-alpha (TNF-α) and interleukin 1 beta (IL-1ß) and interferon-gamma (INF-γ) genes. After A. hydrophila challenge, the intestinal villi epithelium of the L-137/BG group was intact and denser than the other groups. The hepatopancreas and spleen of the control group displayed severe necrosis in hepatocytes and congestion of blood sinusoids in addition to diffuse vacuolation. Regarding the L-137, BG and L-137/BG groups, there was a moderate and normal degree of vacuolation with focal necrosis and mild to moderate degree of congestion of blood sinusoids. Red blood cells, hemoglobin, and albumin showed meaningfully (P < .05) increased values in L-137 or L-137/BG groups. TNF-α, IL-1ß, and INF-γ expressions were upregulated by L-137 and/or BG. The obtained results revealed the ability of L-137 and/or BG to protect Nile tilapia from the effects of A. hydrophila infection by the motivation of the immune, antioxidative, and antiinflammation responses.


Assuntos
Adjuvantes Imunológicos/farmacologia , Aeromonas hydrophila/fisiologia , Doenças dos Peixes/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/veterinária , Lactobacillus plantarum/química , Probióticos/farmacologia , beta-Glucanas/farmacologia , Adjuvantes Imunológicos/administração & dosagem , Ração Animal/análise , Animais , Antioxidantes/metabolismo , Dieta/veterinária , Resistência à Doença/imunologia , Doenças dos Peixes/microbiologia , Proteínas de Peixes/imunologia , Proteínas de Peixes/metabolismo , Infecções por Bactérias Gram-Negativas/tratamento farmacológico , Infecções por Bactérias Gram-Negativas/microbiologia , Probióticos/administração & dosagem , beta-Glucanas/administração & dosagem
16.
Gene ; 742: 144581, 2020 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-32173540

RESUMO

In vertebrates, sex determination and differentiation comprehend a fine balance between female and male factors, leading the bipotential anlage to develop towards ovary or testis, respectively. Nevertheless, the mangrove killifish, (Kryptolebias marmoratus) a simultaneous hermaphroditic species, could overcome those antagonistic pathways and evolved to develop and maintain reproductively active ovarian and testicular tissues in the same organ. Morphological and mRNA localization analyzes of developing and adult gonads demonstrate that genes related to testis (dmrt1 and amh) and ovary differentiation (foxl2 and sox9a) follow the same expression pattern observed in gonochoristic species, thus functioning as two independent organs. In addition, Amh expression patterns make it a strong candidate for initiation of the formation and maintenance of the testicular tissue in the hermaphroditic gonad. Differently from described so far, foxl3 seems to have an important role in oogenesis as well as spermatogenesis and gonadal structure.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Peixes Listrados/fisiologia , Ovário/crescimento & desenvolvimento , Autofertilização/genética , Testículo/crescimento & desenvolvimento , Animais , Feminino , Proteínas de Peixes/metabolismo , Fatores de Transcrição Forkhead/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento/genética , Masculino , Microscopia Eletrônica de Transmissão , Oogênese/genética , Ovário/metabolismo , Ovário/ultraestrutura , Análise Espaço-Temporal , Espermatogênese/genética , Testículo/metabolismo , Testículo/ultraestrutura
17.
Gene ; 742: 144538, 2020 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-32184168

RESUMO

Lysosomal desialylation is the initial step in the degradation of sialo-glycopeptides that is essential for regenerating sialo-glycoconjugates. Neu1 sialidase is the enzyme responsible for the removal of sialic acid in the mammalian lysosome. Although Neu1 sialidases are conserved in fish similar to mammals, their physiological functions remain to be fully understood. Nile tilapia (Oreochromis niloticus) is known to possess two putative Neu1 sialidases (Neu1a and Neu1b) in the genome that may have arisen by gene duplication (specifically in cichlidae family members). This suggests that understanding the Neu1 sialidase in fish, particularly cichlids, could provide insights into the (novel) physiological functions of these genes. Moreover, characterization of the tilapia Neu1 sialidase is paramount to ensure clarity of the desialylation reaction performed by the fish sialidases (like the characterized tilapia sialidases Neu3 and Neu4). Therefore, this study focused on the characterization of the tilapia Neu1 sialidases. Neu1b exhibited narrow substrate specificity when compared with Neu1a, whereas the properties of these two Neu1 sialidases, such as cathepsin A-induced activation, optimal pH, and lysosomal localization, were conserved. Neu1a mRNA levels were detected in various tissues of tilapia as compared to the mRNA levels of Neu1b. Although the cloned construct of Neu1b contained an extra exon unlike tilapia Neu1a, the exon did not affect the enzymatic properties of Neu1b. This study suggests that tilapia Neu1a profiles were highly conserved with other vertebrate Neu1 isoforms, while Neu1b probably evolved independently in other members of the cichlidae family. Moreover, the expression of sialidase genes (neu1a, neu1b, neu3a, and neu4) were determined in various stages of tilapia embryogenesis using real-time PCR; sialidase gene expression is reported to be drastically and individually altered during embryogenesis in Japanese medaka (Oryzias latipes). The mRNA levels of neu1a drastically increased between 72 and 84 hpf and mildly decreased from 84 to 144 hpf. In contrast, the transcript levels of neu1b did not change between 84 and 144 hpf and the expression of neu3a gradually increased between 84 and 120 hpf and drastically decreased at 144 hpf. The highest level of the neu4 transcripts was detected at 84 hpf. These expression patterns were different from those in Japanese medaka, possibly due to the different developmental program found in the tilapia embryo accompanied with the unique profiles of the tilapia sialidases.


Assuntos
Ciclídeos/metabolismo , Proteínas de Peixes/metabolismo , Neuraminidase/metabolismo , Animais , Ciclídeos/genética , Ciclídeos/crescimento & desenvolvimento , Clonagem Molecular , Evolução Molecular , Feminino , Proteínas de Peixes/química , Proteínas de Peixes/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Masculino , Neuraminidase/química , Neuraminidase/genética , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Especificidade por Substrato/genética
18.
Fish Shellfish Immunol ; 98: 595-604, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32004615

RESUMO

Grouper (Epinephelus coioides) is an important commercial maricultural fish, which suffers from nervous necrosis virus (NNV) infection. The molecular mechanisms underlying the pathogenesis of the viral infection are not clear. In this study, we combined deep RNA sequencing and label-free mass spectrum for the first time to analyze the transcriptomic and proteomic profiles in infected/dead, infected/survival (persistent), and infection-free (control)orange-spotted groupers in the larval stage. Further analyses showed that the transcriptome and proteome changed dramatically among the three distinct groups, especially differentially-expressed genes in the infected/dead and infected/survival larvae enriched for pathways related to immune response. Notably, the overlapped genes between transcriptomes and proteomes identified that genes related to collagen synthesis and adhesion molecules were enhanced in the persistent (infected/survival) stage, which might contribute to suppressing the acute and lethal immune responses upon NNV infection. These transcriptomic and proteomic datasets enable the investigation of molecular mechanisms underlying NNV infection, thus may help further development of molecular breeding in marine fishery.


Assuntos
Bass , Moléculas de Adesão Celular/metabolismo , Colágeno/biossíntese , Nodaviridae/fisiologia , Proteoma , Infecções por Vírus de RNA/veterinária , Transcriptoma , Animais , Doenças dos Peixes/metabolismo , Doenças dos Peixes/virologia , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Infecções por Vírus de RNA/metabolismo , Infecções por Vírus de RNA/virologia
19.
Artigo em Inglês | MEDLINE | ID: mdl-32017989

RESUMO

Steroid hormones have been proven as a key drive of sex change in sequentially hermaphroditic organisms. However, the upstream mechanism of sex steroid hormones regulation that affect sex change remain unknown. The main glucocorticoid in teleost fish is cortisol, which both regulates steroidogenesis and has antistress action. Thus, cortisol might be one of the prime factors in sex change. In this study, the glucocorticoid-induced leucine zipper (GILZ) gene, was proven to have a dramatic effect in orange-spotted groupers (Epinephelus coioides) during sex change at the early stage of gonadal transition. The specific action of the GILZ protein is at the pouch-shaped proliferative spermatogonia instead of the degenerative oocyte at the onset of sex change. Immunohistochemical (IHC) evidence revealed that GILZ performs intensively at undifferentiated spermatogonia in the early testis stage. These results imply that cortisol provokes a rise of GILZ through regulation caused by steroid hormones leading to sex change.


Assuntos
Bass/metabolismo , Proteínas de Peixes/metabolismo , Zíper de Leucina/fisiologia , Fatores de Transcrição/metabolismo , Sequência de Aminoácidos , Animais , Bass/genética , Bass/crescimento & desenvolvimento , Feminino , Proteínas de Peixes/biossíntese , Proteínas de Peixes/genética , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Organismos Hermafroditas , Masculino , Filogenia , Homologia de Sequência de Aminoácidos , Diferenciação Sexual/fisiologia , Fatores de Transcrição/biossíntese , Fatores de Transcrição/genética
20.
Sci Adv ; 6(2): eaax4942, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31934625

RESUMO

Through their diet, animals can obtain substances essential for imparting special characteristics, such as toxins in monarch butterflies and luminescent substances in jellyfishes. These substances are typically small molecules because they are less likely to be digested and may be hard for the consumer to biosynthesize. Here, we report that Parapriacanthus ransonneti, a bioluminescent fish, obtains not only its luciferin but also its luciferase enzyme from bioluminescent ostracod prey. The enzyme purified from the fish's light organs was identical to the luciferase of Cypridina noctiluca, a bioluminescent ostracod that they feed upon. Experiments where fish were fed with a related ostracod, Vargula hilgendorfii, demonstrated the specific uptake of the luciferase to the fish's light organs. This "kleptoprotein" system allows an organism to use novel functional proteins that are not encoded in its genome and provides an evolutionary alternative to DNA-based molecular evolution.


Assuntos
Crustáceos/fisiologia , Proteínas de Peixes/metabolismo , Peixes/fisiologia , Luciferases/metabolismo , Medições Luminescentes , Comportamento Predatório/fisiologia , Sequência de Aminoácidos , Estruturas Animais/metabolismo , Animais , Proteínas de Peixes/química , Luciferases/química
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