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1.
PLoS One ; 15(8): e0237111, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32750100

RESUMO

Animal Tubulin-Based-Polymorphism (aTBP), an intron length polymorphism method recently developed for vertebrate genotyping, has been successfully applied to the identification of several fish species. Here, we report data that demonstrate the ability of the aTBP method to assign a specific profile to fish species, each characterized by the presence of commonly shared amplicons together with additional intraspecific polymorphisms. Within each aTBP profile, some fragments are also recognized that can be attributed to taxonomic ranks higher than species, e.g. genus and family. Versatility of application across different taxonomic ranks combined with the presence of a significant number of DNA polymorphisms, makes the aTBP method an additional and useful tool for fish genotyping, suitable for different purposes such as species authentication, parental recognition and detection of allele variations in response to environmental changes.


Assuntos
Proteínas de Peixes/genética , Peixes/genética , Técnicas de Genotipagem/métodos , Polimorfismo Genético , Tubulina (Proteína)/genética , Animais
2.
Gene ; 761: 145037, 2020 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-32777526

RESUMO

Primordial germ cells (PGCs) are singled out from somatic cells very early during embryogenesis, then they migrate towards the genital ridge and differentiate into gametes through oogenesis or spermatogenesis. Labeling PGCs with Localized RNAexpression (LRE) technique by fluorescent proteins has been widely applied among teleost species to study the germ cell development and gonad differentiation. In this study, we first cloned and characterized the 3' untranslated regions (3'UTRs) of nanos homolog 1-like (nos1l), dead end (dnd), and vasa in yellow catfish (Pelteobagrus fulvidraco), and then synthesized the GFP-nos1l/dnd/vasa 3'UTR mRNAs. Each of these three 3'UTRs could label PGCs in yellow catfish embryos, of which, vasa 3'UTR exhibited the highest labeling efficiency. To identify the differences in PGCs at embryonic stage, XX all-female and XY all-male yellow catfish embryos were produced and injected with GFP-vasa 3'UTR mRNA. We observed the PGC migration route in these two monosex embryos from 24 hpf to 7 dpf, and found there was no difference between them. Besides, the PGC number was counted at 48 hpf, and the result showed that the average PGC number in XX females (11.3) was significantly larger than that in XY males (8.1).These findings provide an insight into the development of PGCs in yellow catfish embryos and the relationship between embryonicPGCnumberand thelatergonaddifferentiation.


Assuntos
Peixes-Gato/genética , Gametogênese/genética , Células Germinativas/metabolismo , Regiões 3' não Traduzidas/genética , Sequência de Aminoácidos , Animais , Movimento Celular/genética , RNA Helicases DEAD-box/metabolismo , Desenvolvimento Embrionário/genética , Feminino , Proteínas de Peixes/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Gônadas/metabolismo , Masculino , RNA Mensageiro/genética , Proteínas de Ligação a RNA/metabolismo
3.
PLoS One ; 15(7): e0235617, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32634160

RESUMO

Low egg quality and embryonic survival are critical challenges in aquaculture, where assisted reproduction procedures and other factors may impact egg quality. This includes European eel (Anguilla anguilla), where pituitary extract from carp (CPE) or salmon (SPE) is applied to override a dopaminergic inhibition of the neuroendocrine system, preventing gonadotropin secretion and gonadal development. The present study used either CPE or SPE to induce vitellogenesis in female European eel and compared impacts on egg quality and offspring developmental competence with emphasis on the maternal-to-zygotic transition (MZT). Females treated with SPE produced significantly higher proportions of floating eggs with fewer cleavage abnormalities and higher embryonic survival. These findings related successful embryogenesis to higher abundance of mRNA transcripts of genes involved in cell adhesion, activation of MZT, and immune response (dcbld1, epcam, oct4, igm) throughout embryonic development. The abundance of mRNA transcripts of cldnd, foxr1, cea, ccna1, ccnb1, ccnb2, zar1, oct4, and npm2 was relatively stable during the first eight hours, followed by a drop during MZT and low levels thereafter, indicating transfer and subsequent clearance of maternal mRNA. mRNA abundance of zar1, epcam, and dicer1 was associated with cleavage abnormalities, while mRNA abundance of zar1, sox2, foxr1, cldnd, phb2, neurod4, and neurog1 (before MZT) was associated with subsequent embryonic survival. In a second pattern, low initial mRNA abundance with an increase during MZT and higher levels persisting thereafter indicating the activation of zygotic transcription. mRNA abundance of ccna1, npm2, oct4, neurod4, and neurog1 during later embryonic development was associated with hatch success. A deviating pattern was observed for dcbld1, which mRNA levels followed the maternal-effect gene pattern but only for embryos from SPE treated females. Together, the differences in offspring production and performance reported in this study show that PE composition impacts egg quality and embryogenesis and in particular, the transition from initial maternal transcripts to zygotic transcription.


Assuntos
Anguilla/fisiologia , Carpas/metabolismo , Desenvolvimento Embrionário , Oogênese , Hipófise/metabolismo , Salmão/metabolismo , Anguilla/crescimento & desenvolvimento , Animais , Ciclina A1/genética , Desenvolvimento Embrionário/efeitos dos fármacos , Desenvolvimento Embrionário/genética , Feminino , Proteínas de Peixes/genética , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Fator 3 de Transcrição de Octâmero/genética , Oogênese/efeitos dos fármacos , Hipófise/química , Hormônios Hipofisários/farmacologia , RNA Mensageiro/metabolismo , Zigoto/efeitos dos fármacos , Zigoto/crescimento & desenvolvimento , Zigoto/metabolismo
4.
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
5.
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
6.
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
7.
Mol Immunol ; 125: 83-94, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32652363

RESUMO

Leukocyte immune-type receptors (LITRs) are a multigene family of teleost immunoregulatory proteins that share structural, phylogenetic, and likely functional relationships with several innate immune receptor proteins in other vertebrates, including mammals. Originally discovered in channel catfish (Ictalurus punctatus), representative IpLITR-types have been shown to regulate diverse innate immune cell effector responses including phagocytosis, degranulation, and cytokine secretion. To date, IpLITRs have been primarily characterized using mammalian cell line expression systems, therefore many unanswered questions remain regarding their actual regulatory roles in fish immunity. In the present study, we report on the preliminary molecular characterization of five goldfish (Carassius auratus) CaLITR-types and the identification of several putative splice variants of these receptors cloned from various goldfish tissues and primary myeloid cell cultures. In general, CaLITR mRNA transcripts were detected in all goldfish tissues tested, and also in primary kidney macrophage and neutrophil cultures. Specifically, CaLITR1 is a functionally ambiguous receptor with no charged amino acids in its transmembrane (TM) segment and is devoid of tyrosine-based signaling motifs in its short cytoplasmic tail (CYT) region. CaLITR2 is a putative activating receptor-type that contains immunotyrosine-based activation motifs (ITAMs) within its long CYT region, and CaLITR3 has a positively charged TM segment, suggesting that it may recruit intracellular stimulatory adaptor signaling molecules. CaLITR4 and CaLITR5 appear to have diverse signaling capabilities since they contain various immunoregulatory signaling motifs within their CYT regions including putative Nck and STAT recruitment motifs as well as ITAM-like and ITIM sequences. We also identified putative CaLITR splice variants with altered extracellular Ig-like domain compositions and variable CYT regions. Interestingly, this suggests that alternative splicing-mediated diversification of CaLITRs can generate receptor forms with possible variable binding and/or intracellular signaling abilities. Overall, these findings reveal new information about the teleost LITRs and sets the stage for exploring how alternative splicing leads to the functional diversification of this complex multigene immunoregulatory receptor family.


Assuntos
Carpa Dourada/imunologia , Imunidade Inata/imunologia , Leucócitos/imunologia , Receptores Imunológicos/genética , Receptores Imunológicos/imunologia , Processamento Alternativo , Animais , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Carpa Dourada/genética , Imunidade Inata/genética
8.
Artigo em Inglês | MEDLINE | ID: mdl-32656652

RESUMO

Vibrio alginolyticus is posting an increasing threat to survival of grouper. Classical complement cascade can trigger initiation of immunity, while complement 9 (C9) is a major complement molecule involved in final step of membrane attack complex (MAC) formation. In this study, full-length EcC9 contained an ORF sequence of 1779 bp, encoding a polypeptide of 592 amino acids. A high-level expression of EcC9 mRNA was observed in liver. Following vibrio challenge, increased expression levels of EcC1q, EcBf/C2, EcC4, EcC6, EcC7 and EcC9 mRNA were detected in liver and kidney. These results implied that elevated expression level of classical complement pathway (CCP) and terminal complement components (TCCs) may assess toxicological effect of V. alginolyticus.


Assuntos
Bass/genética , Bass/imunologia , Complemento C9/genética , Complemento C9/imunologia , Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Complemento C9/química , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Filogenia , Alinhamento de Sequência/veterinária , Vibrio alginolyticus/fisiologia
9.
Mol Immunol ; 124: 25-34, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32497752

RESUMO

Tumor necrosis factor receptor superfamily (TNFRSF) is an ancient protein superfamily. By binding to tumor necrosis factor (TNF), it can participate in inflammatory response, apoptosis, lymphocyte homeostasis and tissue development. Seven TNFR members have previously been identified in lampreys but detailed functions of TNFR members are not yet to be resolved. Here, we demonstrate some of the distinguishing features of TNFR10-like member which belongs to TNFRSF. The immunohistochemical results indicate that the TNFR10-like protein is abundant in vascular epithelial cells of the lamprey typhlosole and gills. The expression of tnfr10-like gene has a significantly increased at transcription level after Vibrio anguillarum, Staphylococcus aureus and Poly (I:C) stimulation. Notably, TNFR10-like is specifically expressed in the granulocytes of lamprey peripheral blood and supraneural body. Besides, overexpression tnfr10-like gene in HEK-293 T cells cause a decrease in cell activity and able to activate nuclear transcription factor-κB (NF-κB). Together, these results imply that L-TNFR10-like may play a vital role as a potential marker in lamprey granulocytes and may also be involved in regulation of immune response mediated by itself.


Assuntos
Proteínas de Peixes/imunologia , Granulócitos/imunologia , Lampreias/imunologia , Receptores do Fator de Necrose Tumoral/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Peixes/genética , Células HEK293 , Humanos , Lampreias/genética , Filogenia , Receptores do Fator de Necrose Tumoral/genética
10.
PLoS One ; 15(6): e0234803, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32544202

RESUMO

The genetic bases of growth and body weight are of economic and scientific interest, and teleost fish models have proven useful in such investigations. The Oryzias latipes species complex (medaka) is an abundant freshwater fish in Japan and suitable for genetic studies. We compared two wild medaka stocks originating from different latitudes. The Maizuru population from higher latitudes weighed more than the Ginoza population. We investigated the genetic basis of body weight, using quantitative trait locus (QTL) analysis of the F2 offspring of these populations. We detected one statistically significant QTL for body weight on medaka chromosome 4 and identified 12 candidate genes that might be associated with body weight or growth. Nine of these 12 genes had at least one single nucleotide polymorphism that caused amino acid substitutions in protein-coding regions, and we estimated the effects of these substitutions. The present findings might contribute to the marker-assisted selection of economically important aquaculture species.


Assuntos
Peso Corporal/genética , Variação Genética , Oryzias/genética , Sequência de Aminoácidos , Animais , Cromossomos/genética , Proteínas de Peixes/química , Proteínas de Peixes/genética , Oryzias/fisiologia , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Alinhamento de Sequência
11.
PLoS One ; 15(6): e0234377, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32542044

RESUMO

In many aquatic species, alteration of habitats and human-induced barriers shape the population's genetic structure in rivers with longitudinal connectivity. The golden mahseer, Tor putitora (GM) is an endangered and sensitive cyprinid species. It is considered an indicator of a healthy freshwater ecosystem and is found in cold-water habitats. Therefore, it is crucial to understand how longitudinal connectivity and anthropogenic factors affect the diversity and population genetic structure of GM. The population genetic structure, gene flow and demography of the GM in four Himalayan rivers were investigated by mitochondrial cytochrome b gene (cyt b) as well as microsatellite genotyping. The results showed overall high mtDNA diversity (hd: 0.795) couple with low nucleotide diversity (π: 0.0012) in all GM populations. We also found significant levels of observed heterozygosity (ranging from 0.618 to 0.676), with three genetic clusters. The mtDNA and microsatellite analysis suggested that there are close genetic relationships between the Bhagirathi and Ganga populations; whereas, significant level of genetic differentiation was observed with that of Alaknanda and Yamuna populations. Haplotype distribution, unimodal distribution graph and results of the neutrality test indicated a sign of recent population growth in the GM population. Analysis of molecular variance (AMOVA) and spatial molecular variance (SAMOVA) revealed existence of genetic structures in GM populations. In addition, spatial genetic analysis detected a significant correlation between the pairwise genetic and geographical distances for the entire study area (Mantel test, rM = 0.126; P = 0.010). Considering the significant level of heterozygosity, high rate of unidirectional migration and the intra-population structuring in Alaknanda and Yamuna, it is crucial to propose an effective conservation plan for the GM populations. In general, dams obstruct continuous water flow and create isolated microhabitats. Therefore, we recommend the establishment of microscale protected areas near GM breeding sites and construction of fish pass to maintain the genetic connectivity of fish species that enhance viable populations.


Assuntos
Cyprinidae/genética , Distribuição Animal , Migração Animal , Animais , Conservação dos Recursos Naturais , Cyprinidae/classificação , Citocromos b/genética , DNA Mitocondrial/genética , Ecossistema , Espécies em Perigo de Extinção , Proteínas de Peixes/genética , Fluxo Gênico , Genes Mitocondriais , Variação Genética , Genética Populacional , Haplótipos , Heterozigoto , Índia , Repetições de Microssatélites , Filogeografia , Polimorfismo Genético , Rios
12.
Gene ; 753: 144777, 2020 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-32428695

RESUMO

As a crucial member of the Forkhead Box family, class O (FoxO) plays an essential role in growth, cell differentiation, metabolism, immunization, and apoptosis. Meanwhile, FoxO3 is the primary regulator and effective inhibitor of primordial follicle activation. In this study, seven foxo genes were identified in black rockfish (Sebastes schlegelii), including two foxo1 genes (foxo1a, foxo1b), two foxo3 genes (foxo3, foxo3l), one foxo4 gene, and two foxo6 genes (foxo6a, foxo6b). foxo3l was derived from teleost-specific whole-genome duplication events. Evaluation of tissue expression pattern revealed that foxo3l displayed sexually dimorphic expression with a high level in the ovary and spatial expression only in the cytoplasm of follicle cells and oocytes. When the ovaries were stimulated by estrogen and gonadotropin, foxo3l expression was remarkably reduced, and the effect of androgen was completely different. We considered that foxo3l lost its ability to inhibit follicular precocity because of mass ovulation by hormone stimulation, resulting in its decreased expression. Such evidence indicated that foxo3l is an important regulator of reproduction-related functions in black rockfish. This study provides new insights into foxo3l genes for further functional research in teleost.


Assuntos
Proteína Forkhead Box O3/genética , Perciformes/genética , Sequência de Aminoácidos , Animais , Feminino , Doenças dos Peixes/imunologia , Proteínas de Peixes/genética , Proteína Forkhead Box O3/metabolismo , Fatores de Transcrição Forkhead/genética , Duplicação Gênica/genética , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica/genética , Imunidade Inata/genética , Oócitos/metabolismo , Oogênese/genética , Ovário/metabolismo , Filogenia , Alinhamento de Sequência
13.
PLoS One ; 15(5): e0233081, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32463827

RESUMO

ZMAT2 is among the least-studied of mammalian proteins and genes, even though it is the ortholog of Snu23, a protein involved in pre-mRNA splicing in yeast. Here we have used data from genomic and gene expression repositories to examine the Zmat2 gene and locus in 8 terrestrial vertebrates, 10 ray-finned fish, and 1 lobe-finned fish representing > 500 million years of evolutionary diversification. The analyses revealed that vertebrate Zmat2 genes are similar to their mammalian counterparts, as in 16/19 species studied they contain 6 exons, and in 18/19 encode a single conserved protein. However, unlike in mammals, no Zmat2 pseudogenes were identified in these vertebrates, although an expressed Zmat2 paralog was characterized in flycatcher that resembled a DNA copy of a processed and retro-transposed mRNA, and thus could be a proto-pseudogene captured during its evolutionary journey from active to inert. The Zmat2 locus in terrestrial vertebrates, and in spotted gar and coelacanth, also shares additional genes with its mammalian counterparts, including Histidyl-tRNA synthetase (Hars), Hars2, and others, but these are absent from the Zmat2 locus in teleost fish, in which Stem-loop-binding protein 2 (Slbp2) and Lymphocyte cytosolic protein 2a (Lcp2a) are present instead. Taken together, these observations argue that a recognizable Zmat2 was present in the earliest vertebrate ancestors, and postulate that during chromosomal tetraploidization and subsequent re-diploidization during modern teleost evolution, the duplicated Zmat2 gene was retained and the original lost. This study also highlights how information from genomic resources can be leveraged to reveal new biologically significant insights.


Assuntos
Proteínas de Peixes/genética , Peixes/classificação , Sequência de Aminoácidos , Animais , Sequência Conservada , Evolução Molecular , Proteínas de Peixes/química , Peixes/genética , Duplicação Gênica , Filogenia , Dedos de Zinco
14.
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
15.
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
16.
Science ; 368(6492): 731-736, 2020 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-32409469

RESUMO

The establishment of reproductive barriers between populations can fuel the evolution of new species. A genetic framework for this process posits that "incompatible" interactions between genes can evolve that result in reduced survival or reproduction in hybrids. However, progress has been slow in identifying individual genes that underlie hybrid incompatibilities. We used a combination of approaches to map the genes that drive the development of an incompatibility that causes melanoma in swordtail fish hybrids. One of the genes involved in this incompatibility also causes melanoma in hybrids between distantly related species. Moreover, this melanoma reduces survival in the wild, likely because of progressive degradation of the fin. This work identifies genes underlying a vertebrate hybrid incompatibility and provides a glimpse into the action of these genes in natural hybrid populations.


Assuntos
Ciprinodontiformes/genética , Doenças dos Peixes/genética , Proteínas de Peixes/genética , Hibridização Genética , Melanoma/genética , Melanoma/virologia , Receptores Proteína Tirosina Quinases/genética , Alelos , Nadadeiras de Animais/patologia , Animais , Quimera , Loci Gênicos , Estudo de Associação Genômica Ampla
17.
Arch Virol ; 165(7): 1599-1609, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32399788

RESUMO

The viral hemorrhage disease caused by grass carp reovirus (GCRV) is a serious contagious disease of grass carp that mainly infects fingerlings and yearlings. Epidemiological studies have shown that GCRV genotype II is currently the prominent genotype. However, little is known about the histopathological characteristics, virus distribution, and expression of immunity-related genes in grass carp infected by GCRV genotype II. In this study, we found that grass carp infected by GCRV genotype II lost appetite, swam alone, and rolled, and their fins, eyes, operculum, oral cavity, abdomen, intestine, and muscles showed pronounced punctate hemorrhage. Congestion, swelling, deformation, thinning of membranes, dilatation and darkened color of nucleoli, cathepsis, erythrocyte infiltration, and vacuole formation were observed in some infected tissues. A qRT-PCR test showed that the 11 genome segments of GCRV had similar expression patterns in different tissues. The S8 segment, with unknown function and no homologous sequences, had the highest expression level, while the most conserved segment, L2, had the lowest expression level. GCRV particles were distributed in different tissues, especially in the intestine. In the infected intestine, the expression of various receptors and adaptor molecules was modulated at different levels. Pro-inflammatory cytokine interleukin-1ß (IL-1ß) expression was 2160.9 times higher than that in the control group. The upregulation of immunity-related genes activated the antiviral immunity pathways. Therefore, the intestine might play a dual role in mediating GCRV infection and the antiviral immune response. This study provides detailed information about the pathogenicity of GCRV and expression of immunity-related genes, laying the foundation for further research on virus control and treatment.


Assuntos
Carpas/virologia , Doenças dos Peixes/genética , Proteínas de Peixes/genética , Infecções por Reoviridae/veterinária , Reoviridae/fisiologia , Animais , Carpas/genética , Carpas/imunologia , Doenças dos Peixes/imunologia , Doenças dos Peixes/patologia , Doenças dos Peixes/virologia , Proteínas de Peixes/imunologia , Genoma Viral , Genótipo , Interleucina-1beta/genética , Interleucina-1beta/imunologia , Reoviridae/genética , Infecções por Reoviridae/genética , Infecções por Reoviridae/imunologia , Infecções por Reoviridae/virologia
18.
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
19.
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
20.
Gene ; 745: 144628, 2020 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-32224271

RESUMO

microRNAs (miRNAs) are short, endogenous non-coding RNAs that contain approximately 18-22 nucleotides. miRNAs are involved in gene regulation by recognizing and binding the 3'UTR of target gene. In our previous data, miR-430 family showed significant differential expression modes through metamorphosis in Japanese flounder. It was speculated that miR-430a plays a key role in left-right patterning. We predicted the targets of miR-430a and gene ontology (GO) was performed. We speculated miR-430a is involved in the basal molecular function and organ development. In Japanese flounder, sqt as a target of miR-430a was enriched into heart development term. Sqt has been reported to participate in mesendoderm formation and organ development. Cardiac morphogenesis is the first asymmetric development process, which breaks left-right symmetry in bilateria. It was used as a marker to detect L-R asymmetric effects of miR-430a. Overexpression and suppression of miR-430a resulted in abnormal KV (Kupffer's vesicles) development and disordered in nodal-related expression with consequent cardiac laterality. Squint mRNA of Japanese flounder (Posqt) as a target of miR-430a was overexpressed and caused similar phenotype with miR-430a suppression group, such as longer cilia in KV and high range of clmc2 and spaw ectopic expression. Moreover, rescue experiments were performed and suggested that cardiac and KV defections, induced by overexpressing miR-430a, could be rescued by injecting Posqt mRNA. These results suggested that miR-430a regulates the development of left-right asymmetry by targeting sqt in the teleost.


Assuntos
Padronização Corporal/genética , Proteínas de Peixes/genética , Linguado/genética , Regulação da Expressão Gênica no Desenvolvimento , MicroRNAs/metabolismo , Animais , Embrião não Mamífero , Linguado/crescimento & desenvolvimento , Coração/crescimento & desenvolvimento , Metamorfose Biológica , Peixe-Zebra
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