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1.
Biol Sex Differ ; 15(1): 70, 2024 Sep 07.
Artículo en Inglés | MEDLINE | ID: mdl-39244546

RESUMEN

BACKGROUND: Blotched snakehead (Channa maculata) displays significant sexual dimorphism, with males exhibiting faster growth rates and larger body sizes compared to females. The cultivation of the all-male population of snakeheads holds substantial economic and ecological value. Nonetheless, the intricate processes governing the development of bipotential gonads into either testis or ovary in C. maculata remain inadequately elucidated. Therefore, it is necessary to determine the critical time window of sex differentiation in C. maculata, providing a theoretical basis for sex control in production practices. METHODS: The body length and weight of male and female C. maculata were measured at different developmental stages to reveal when sexual dimorphism in growth initially appears. Histological observations and spatiotemporal comparative transcriptome analyses were performed on ovaries and testes across various developmental stages to determine the crucial time windows for sex differentiation in each sex and the sex-related genes. Additionally, qPCR and MG2C were utilized to validate and locate sex-related genes, and levels of E2 and T were quantified to understand sex steroid synthesis. RESULTS: Sexual dimorphism in growth became evident starting from 90 dpf. Histological observations revealed that morphological sex differentiation in females and males occurred between 20 and 25 dpf or earlier and 30-35 dpf or earlier, respectively, corresponding to the appearance of the ovarian cavity or efferent duct anlage. Transcriptome analyses revealed divergent gene expression patterns in testes and ovaries after 30 dpf. The periods of 40-60 dpf and 60-90 dpf marked the initiation of molecular sex differentiation in females and males, respectively. Male-biased genes (Sox11a, Dmrt1, Amh, Amhr2, Gsdf, Ar, Cyp17a2) likely play crucial roles in male sex differentiation and spermatogenesis, while female-biased genes (Foxl2, Cyp19a1a, Bmp15, Figla, Er) could be pivotal in ovarian differentiation and development. Numerous biological pathways linked to sex differentiation and gametogenesis were also identified. Additionally, E2 and T exhibited sexual dimorphism during sex differentiation and gonadal development. Based on these results, it is hypothesized that in C. maculata, the potential male sex differentiation pathway, Sox11a-Dmrt1-Sox9b, activates downstream sex-related genes (Amh, Amhr2, Gsdf, Ar, Cyp17a2) for testicular development, while the antagonistic pathway, Foxl2/Cyp19a1a, activates downstream sex-related genes (Bmp15, Figla, Er) for ovarian development. CONCLUSIONS: This study provides a comprehensive overview of gonadal dynamic changes during sex differentiation and gametogenesis in C. maculata, establishing a scientific foundation for sex control in this species.


Blotched snakehead (Channa maculata) exhibits significant sexual dimorphism, as males display faster growth rates and larger body sizes compared to females. The cultivation of the all-male population of snakeheads holds substantial economic and ecological value. However, the mechanisms underlying sex determination and differentiation in C. maculata remain insufficiently elucidated. In this study, sexual dimorphism in growth became evident starting from 90 dpf through the measurement of body length and weight of male and female C. maculata at different developmental stages. Histological observations indicated that morphological sex differentiation in females and males occurred at 20­25 dpf or earlier and 30­35 dpf or earlier, respectively, corresponding to the appearance of the ovarian cavity or efferent duct anlage. Transcriptome analyses revealed divergent gene expression patterns in male and female gonads after 30 dpf, suggesting that the period preceding 30 dpf might be the critical time window for sex control in C. maculata. The periods of 40­60 dpf and 60­90 dpf marked the initiation of molecular sex differentiation in females and males, respectively. Male-biased genes (Sox11a, Dmrt1, Amh, Amhr2, Gsdf, Ar, Cyp17a2) likely play crucial roles in testicular differentiation and spermatogenesis, while female-biased genes (Foxl2, Cyp19a1a, Bmp15, Figla, Er) could be pivotal in ovarian differentiation and oogenesis. Additionally, numerous biological pathways linked to sex differentiation and gametogenesis were identified. Moreover, sexual dimorphism was observed in the levels of E2 and T during gonadal differentiation and development. Based on these findings, it is hypothesized that in C. maculata, the potential male sex differentiation pathway, Sox11a­Dmrt1­Sox9b, activates downstream sex-related genes (Amh, Amhr2, Gsdf, Ar, Cyp17a2) for testicular development, while the antagonistic pathway, Foxl2/Cyp19a1a, activates downstream sex-related genes (Bmp15, Figla, Er) for ovarian development. This study provides a comprehensive overview of gonadal dynamic changes during sex differentiation and gametogenesis in C. maculata, thereby establishing a scientific foundation for sex control in this species.


Asunto(s)
Gametogénesis , Caracteres Sexuales , Diferenciación Sexual , Animales , Femenino , Masculino , Gónadas/crecimiento & desarrollo , Gónadas/anatomía & histología , Perfilación de la Expresión Génica , Peces/crecimiento & desarrollo , Peces/anatomía & histología , Peces/genética , Transcriptoma , Testículo/crecimiento & desarrollo , Testículo/anatomía & histología , Ovario/crecimiento & desarrollo , Ovario/anatomía & histología , Regulación del Desarrollo de la Expresión Génica , Channa punctatus
2.
Fish Shellfish Immunol ; 149: 109555, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38615703

RESUMEN

Developing a low-protein feed is important for the sustainable advancement of aquaculture. The aim of this study was to investigate the effects of essential amino acid (EAA) supplementation in a low-protein diet on the growth, intestinal health, and microbiota of the juvenile blotched snakehead, Channa maculata in an 8-week trial conducted in a recirculating aquaculture system. Three isoenergetic diets were formulated to include a control group (48.66 % crude protein (CP), HP), a low protein group (42.54 % CP, LP), and a low protein supplementation EAA group (44.44 % CP, LP-AA). The results showed that significantly lower weight gain (WG), specific growth rate (SGR), protein efficiency ratio (PER), and feed efficiency ratio (FER) were observed in fish that were fed LP than in the HP and LP-AA groups (P < 0.05). The HP and LP-AA groups exhibited a significant increase in intestinal villus length, villus width, and muscular thickness compared to the LP group (P < 0.05). Additionally, the HP and LP-AA groups demonstrated significantly higher levels of intestinal total antioxidant capacity (T-AOC), catalase (CAT), and superoxide dismutase (SOD) and lower levels of malondialdehyde (MDA) compared to the LP group (P < 0.05). The apoptosis rate of intestinal cells in the LP group was significantly higher than those in the LP and HP groups (P < 0.05). The mRNA expression levels of superoxide dismutase (sod), nuclear factor kappa B p65 subunit (nfκb-p65), heat shock protein 70 (hsp70), and inhibitor of NF-κBα (iκba) in the intestine were significantly higher in the LP group than those in the HP and LP-AA groups (P < 0.05). The 16s RNA analysis indicated that EAA supplementation significantly increased the growth of Desulfovibrio and altered the intestinal microflora. The relative abundances of Firmicutes and Cyanobacteria were positively correlated with antioxidant parameters (CAT and T-AOC), whereas Desulfobacterota was negatively correlated with sod and T-AOC. The genera Bacillus, Bacteroides, and Rothia were associated with the favorable maintenance of gut health. In conclusion, dietary supplementation with EAAs to achieve a balanced amino acid profile could potentially reduce the dietary protein levels from 48.66 % to 44.44 % without adversely affecting the growth and intestinal health of juvenile blotched snakeheads.


Asunto(s)
Aminoácidos Esenciales , Alimentación Animal , Suplementos Dietéticos , Microbioma Gastrointestinal , Intestinos , Animales , Alimentación Animal/análisis , Suplementos Dietéticos/análisis , Microbioma Gastrointestinal/efectos de los fármacos , Aminoácidos Esenciales/administración & dosificación , Perciformes/crecimiento & desarrollo , Perciformes/inmunología , Dieta con Restricción de Proteínas/veterinaria , Dieta/veterinaria , Distribución Aleatoria , Peces/crecimiento & desarrollo , Acuicultura , Channa punctatus
3.
Animals (Basel) ; 14(3)2024 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-38338134

RESUMEN

Figla is one of the earliest expressed genes in the oocyte during ovarian development. In this study, Figla was characterized in C. maculata, one of the main aquaculture species in China, and designated as CmFigla. The length of CmFigla cDNA was 1303 bp, encoding 197 amino acids that contained a conserved bHLH domain. CmFigla revealed a female-biased expression patterns in the gonads of adult fish, and CmFigla expression was far higher in ovaries than that in testes at all gonadal development stages, especially at 60~180 days post-fertilization (dpf). Furthermore, a noteworthy inverse relationship was observed between CmFigla expression and the methylation of its promoter in the adult gonads. Gonads at 90 dpf were used for in situ hybridization (ISH), and CmFigla transcripts were mainly concentrated in oogonia and the primary oocytes in ovaries, but undetectable in the testes. These results indicated that Figla would play vital roles in the ovarian development in C. maculata. Additionally, the frame-shift mutations of CmFigla were successfully constructed through the CRISPR/Cas9 system, which established a positive foundation for further investigation on the role of Figla in the ovarian development of C. maculata. Our study provides valuable clues for exploring the regulatory mechanism of Figla in the fish ovarian development and maintenance, which would be useful for the sex control and reproduction of fish in aquaculture.

4.
Biology (Basel) ; 12(10)2023 Oct 12.
Artículo en Inglés | MEDLINE | ID: mdl-37887041

RESUMEN

Based on obtaining mstnb gene knockout in Pelteobagrus fulvidraco, a study on the effect of the mstn gene on skeletal morphology and growth was performed by comparing the number and length of the vertebrae of mutant and wild-type fish in a sibling group of P. fulvidraco, combined with the differences in cells at the level of vertebral skeletal tissue. It was found that mstnb gene knockdown resulted in a reduction in the number of vertebrae, the length, and the intervertebral distance in P. fulvidraco, and these changes may be the underlying cause of the shorter body length in mutant P. fulvidraco. Further, histological comparison of the same sites in the mstn mutant and wild groups of P. fulvidraco also revealed that the number and density of osteocytes were greater in mstnb knockout P. fulvidraco than in wild-type P. fulvidraco. Our results demonstrated that when using genome editing technology to breed new lines, the effects of knockout need to be analyzed comprehensively and may have some unexpected effects due to insufficient study of the function of certain genes.

5.
Fish Physiol Biochem ; 49(6): 1215-1227, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37857788

RESUMEN

Many fish species exhibit remarkable sexual dimorphism, with males possessing numerous advantageous traits for commercial production by aquaculture such as faster growth rate, more efficient food energy utilization for muscle development, and better breeding performance. Several studies have shown that a decrease in the number of primordial germ cells (PGCs) during early development leads predominantly to male progeny. In this study, we developed a method to obtain all-male zebrafish (Danio rerio) by targeted PGC ablation using the nitroreductase/metronidazole (NTR/Mtz) system. Embryos generated by female heterozygous Tg(nanos3:nfsB-mCherry-nanos3 3'UTR) and male wild-types (WTs) were treated with vehicle or Mtz. Compared to vehicle-treated controls, 5.0 and 10.0 mM Mtz treatment for 24 h significantly reduced the number of PGCs and yielded an exclusively male phenotype in adulthood. The gonads of offspring treated with 5.0 mM Mtz exhibited relatively normal morphology and histological characteristics. Furthermore, these males were able to chase females, spawn, and produce viable offspring, while about 20.0% of males treated with 10.0 mM Mtz were unable to produce viable offspring. The 5.0 mM Mtz treatment protocol may thus be suitable for large-scale production of fertile male offspring. Moreover, about half of these males were WT as evidenced by the absence of nfsB gene expression. It may thus be possible to breed an all-male WT fish population by Mtz-mediated PGC ablation.


Asunto(s)
Perciformes , Pez Cebra , Animales , Masculino , Femenino , Pez Cebra/fisiología , Proteínas de Pez Cebra/genética , Células Germinativas , Fertilidad , Perciformes/metabolismo
6.
Artículo en Inglés | MEDLINE | ID: mdl-34455080

RESUMEN

Dmrt1 is an important transcriptional regulator that plays critical role in male gonadogenesis, testicular differentiation and development. In this study, Dmrt1 was cloned from blotched snakehead (Channa maculata), which is designated as CmDmrt1. CmDmrt1 encoded a putative protein with 293 amino acids and presented an extremely conserved DM domain. It was nearly expressed in the gonads, and the expression was more than 15 times higher in the testis than in the ovary. 1851 bp promoter sequence of CmDmrt1 was characterized and the methylation levels of the CpG sites were analyzed to detect sex-related differences. A significant negative correlation between CmDmrt1 expression and CpG methylation level of its promoter was found in the testis and ovary. During gonadal development, CmDmrt1 transcription displayed strong male-biased expression patterns, increased with the maturation of testis and reached the peak at 195 days after hatching (dah), which indicates a significant role of Dmrt1 in spermatogenesis. Steroid treatment could influence CmDmrt1 expression, and long-term 17ß-estradiol (E2) treatment could induce the male-to-female secondary sex reversal (SSR), which resulted in the differentiated testis transformed to ovary or ovotestis. Meanwhile, CmDmrt1 expression was down-regulated to fairly low level in the ovary of the SSR XY fish, which was similar to that in normal XX females ovary. Our research illustrates that Dmrt1 is linked to testis differentiation and spermatogenesis in blotched snakehead, providing information for functional studies on sex differentiation and gonadal development of C. maculata, and scientific basis for the production practice of all-male snakehead breeding.


Asunto(s)
Gónadas , Diferenciación Sexual , Animales , Estradiol/metabolismo , Estradiol/farmacología , Femenino , Peces/genética , Regulación del Desarrollo de la Expresión Génica , Gónadas/metabolismo , Masculino , Metilación , Diferenciación Sexual/genética , Testículo/metabolismo
7.
Gigascience ; 10(10)2021 10 21.
Artículo en Inglés | MEDLINE | ID: mdl-34673930

RESUMEN

BACKGROUND: Channa argus and Channa maculata are the main cultured species of the snakehead fish family, Channidae. The relationship between them is close enough that they can mate; however, their temperature adaptability is quite different. RESULTS: In this study, we sequenced and assembled the whole genomes of C. argus and C. maculata and obtained chromosome-level genome assemblies of 630.39 and 618.82 Mb, respectively. Contig N50 was 13.20 and 21.73 Mb, and scaffold N50 was 27.66 and 28.37 Mb, with 28,054 and 24,115 coding genes annotated for C. argus and C. maculata, respectively. Our analyses showed that C. argus and C. maculata have 24 and 21 chromosomes, respectively. Three pairs of chromosomes in C. argus correspond to 3 chromosomes in C. maculata, suggesting that 3 chromosomal fusion events occurred in C. maculata. Comparative analysis of their gene families showed that some immune-related genes were unique or expandable to C. maculata, such as genes related to herpes simplex infection. Analysis of the transcriptome differences related to temperature adaptation revealed that the brain and liver of C. argus rapidly produced more differentially expressed genes than C. maculata. Genes in the FoxO signalling pathway were significantly enriched in C. argus during the cooling process (P < 0.05), and the expression of 3 transcription factor genes in this pathway was significantly different between C. argus and C. maculata (P < 0.01). CONCLUSIONS: C. maculata may have higher resistance to certain diseases, whereas C. argus has a faster and stronger response to low-temperature stress and thus has better adaptability to a low-temperature environment. This study provides a high-quality genome research platform for follow-up studies of Channidae and provides important clues regarding differences in the low-temperature adaptations of fish.


Asunto(s)
Cromosomas , Peces , Animales , Cromosomas/genética , Peces/genética , Genoma , Temperatura , Transcriptoma
8.
Artículo en Inglés | MEDLINE | ID: mdl-32674038

RESUMEN

Blotched snakehead (Channa maculata) is an economically important freshwater fish in China, of which males grow much faster than females. To illuminate the molecular mechanism of sex differentiation and gonad development, RNA-Sequencing was performed to identify sex-related genes and pathway in gonads of 6-month-old normal XX females (XX-F), normal XY males (XY-M), XY sex reversal females (XY-F) and YY super-males (YY-M). The analysis showed that many differentially expressed genes (DEGs) had similar expression patterns in XY-F and XX-F, which were different from XY-M and YY-M. qRT-PCR indicated that Amh, Dmrt1, and Sox9 had relatively high expression in testes of XY-M and YY-M. Taking Amh as an example, there was a relative fold change of 1.0 in XX-F, 2.1 fold change in XY-F, 36.1 fold change in XY-M, and 26.0 fold change in YY-M. Cyp19a1a, Figla, and Foxl2 were highly expressive in ovaries of XX-F and XY-F. Taking Figla as an example, there was a relative fold change of 557 in XX-F, 304.5 fold change in XY-F, 5.6 fold change in XY-M, and 4.4 fold change in YY-M. KEGG analysis revealed many DEGs distributed in pathways related to sex differentiation, steroid hormone synthesis and growth, etc. Significant variation and trends in relative expression levels tested by qRT-PCR were consistent with those recorded by RNA-Sequencing. This is the first time that transcriptome of snakehead has been investigated systematically and in an integrated way. Large quantities of candidate genes involved in sex differentiation, gonad development and growth dimorphism were identified. The study provides useful resources for understanding sex differentiation and growth dimorphism, potentially assisting mono-sex production of snakehead in aquaculture.


Asunto(s)
Peces/genética , Transcriptoma , Animales , Femenino , Peces/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica , Ontología de Genes , Gónadas/metabolismo , Gónadas/ultraestructura , Masculino , Diferenciación Sexual
9.
Genes Genomics ; 42(7): 827-835, 2020 07.
Artículo en Inglés | MEDLINE | ID: mdl-32462521

RESUMEN

BACKGROUND: Anti-Müllerian hormone (Amh) plays a critical role in both early sex determination and later gonad development in vertebrate species. However, it remains unknown in northern snakehead (Channa argus), which is economically important freshwater fish with sexual dimorphism. OBJECTIVE: This study aimed to identify the expression profiles and estrogenic regulation of CaAmh during gonadal sex differentiation in C. argus. METHODS: The cDNA and genomic DNA sequences of CaAmh were identified by PCR and RACE techniques. The expression patterns of CaAmh were detected by qRT-PCR during the gonadal sex differentiation and after 17α-ethinyloestradiol (EE2) treatments. RESULTS: CaAmh is composed of seven exons and six introns, and its full-length cDNA is 2413 bp in length, with 1635 bp open reading frame (ORF) that encodes a 544 amino acid protein. Tissues expression patterns revealed that CaAmh display the highest expression in testis of XY males (40.36 folds, p < 0.01). The spatio-temporal expression patterns during gonadal sex differentiation indicated that CaAmh expression differed between XX females and XY males at 30 day after hatching (dah), and reached to the peak (36.03 folds, p < 0.01) at 90 dah in XY gonads. However, CaAmh expression in XX gonads remained low throughout the sampling period. Furthermore, CaAmh expression in the gonads (ovaries) of the sex-reversed XY fish (XY-F) by the administration of estrogen EE2 was downregulated to low level, similar to that in ovaries of normal XX females (XX-F). CONCLUSIONS: These results show that Amh plays a critical role in testicular differentiation of C. argus and it is apparently modulated by estrogens in this species.


Asunto(s)
Hormona Antimülleriana/genética , Etinilestradiol/farmacología , Peces/genética , Ovario/metabolismo , Diferenciación Sexual/efectos de los fármacos , Testículo/metabolismo , Animales , Femenino , Peces/crecimiento & desarrollo , Peces/fisiología , Regulación del Desarrollo de la Expresión Génica , Masculino , Ovario/efectos de los fármacos , Ovario/fisiología , Testículo/efectos de los fármacos , Testículo/fisiología
10.
Gene ; 703: 125-133, 2019 Jun 30.
Artículo en Inglés | MEDLINE | ID: mdl-30978477

RESUMEN

Hybrid vigour, or heterosis, refers to the increased productivity and growth rate of hybrid offsprings relative to the parents. Various heterosis have been well exploited in fish for fisheries. However, the molecular mechanisms underlying heterosis are largely unknown in fish. In this study, two inbred and hybrid lines between the northern snakehead (NS, Channa argus) and blotched snakehead (BS, Channa maculata) were generated. The analysis on various growth traits, including body length, head length, and body height, showed that hybrid fry obviously exhibited a spontaneous growth heterosis over the inbred. Moreover, the methylation-sensitive amplification polymorphism (MSAP) analysis revealed that the DNA methylation levels were negatively related to the body growth in all fry. Especially, the DNA methylation levels in the hybrid fry were significantly lower than those in the inbred. Additionally, qRT-PCR showed that the snakehead fish Dnmt3a mRNA was initially detectable in embryos at 12 hpf and gradually increased as developing. Intriguingly, the level of Dnmt3a mRNA expression was found to be closely correlated to the DNA methylation level in embryos/fry. The results of this study firstly demonstrated the correlations between growth heterosis, DNA methylation level and Dnmt3a mRNA expression in fish fry. The findings of this study implied that the hybrids' heterosis formation is probably accompanied by DNA methylation alterations and modulated by Dnmt3a gene in fish. This study would provide new clues for further investigations on mechanisms behind heterosis formation in fish hybrid.


Asunto(s)
ADN (Citosina-5-)-Metiltransferasas/genética , Metilación de ADN , Peces/crecimiento & desarrollo , Animales , Cruzamiento , ADN Metiltransferasa 3A , Femenino , Peces/genética , Regulación del Desarrollo de la Expresión Génica , Vigor Híbrido , Masculino , Sitios de Carácter Cuantitativo
11.
Fish Shellfish Immunol ; 89: 614-622, 2019 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-30991152

RESUMEN

Scavenger receptor class B type 1 (SRB1) is a transmembrane protein belonging to the scavenger receptors (SRs) family and it plays an important role in viral entry. Not much is known on SRB1 in teleost fish. Grass carp reovirus (GCRV) cause huge economic losses in grass carp industry. In this study, rare minnow (Gobiocypris rarus) was used as a model fish to investigate the mechanism of GCRV infection, which is sensitive to GCRV. The structure of SRB1 gene in G. rarus (GrSRB1) was cloned and elucidated. GrSRB1 is composed of 13 exons and 12 introns, and its full-length cDNA is 2296 bp in length, with 1521 bp open reading frame (ORF) that encodes a 506 amino acid protein. The GrSRB1 protein is predicted to contain a typical CD36 domain and two transmembrane regions. In G. rarus, GrSRB1 is expressed strongly in the liver (L), intestines (I), brain (B) and muscle (M), while it is expressed poorly in the heart (H), middle kidney (MK), head kidney (HK) and gills (G). After infection with GCRV, GrSRB1 expression was up-regulated in main immune tissues during the early infection period. Moreover, co-immunoprecipitation assays revealed that GrSRB1 could interact with the outer capsid protein of GCRV (VP5 and VP7). These results suggest that GrSRB1 could be a receptor for GCRV. We have managed to characterize the GrSRB1 gene and provide evidence for its potential functions for GCRV entry into host cells.


Asunto(s)
Antígenos CD36/genética , Antígenos CD36/inmunología , Cyprinidae/genética , Cyprinidae/inmunología , Enfermedades de los Peces/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Antígenos CD36/química , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica/veterinaria , Filogenia , Reoviridae/fisiología , Infecciones por Reoviridae/inmunología , Infecciones por Reoviridae/veterinaria , Alineación de Secuencia/veterinaria
12.
Oncotarget ; 8(58): 98733-98744, 2017 Nov 17.
Artículo en Inglés | MEDLINE | ID: mdl-29228723

RESUMEN

We described a next generation sequencing (NGS)-based approach to identify sex-specific markers and subsequently determine whether a species has male or female heterogamety. To test the accuracy of this technique, we examined the snakehead (Channa argus), which is economically important freshwater fish in China. Males grow faster than females, and there is significant interest in developing methods to skew breeding towards all-males to increase biomass yields. NGS was conducted on DNAs of individual female and male, the male reads were spitted into 60 bp K-mers and aligned to the female reference genome assembled by female reads, unaligned male K-mers-60 were kept in next filter process. Meanwhile, DNA sample of 48 females was pooled and sequenced, this data was further used to filter out the previous unaligned male K-mers-60. Hence, numbers of candidate Y chromosome-specific sequences were screened out, their sex-specificity were validated in wild snakeheads through PCR amplification. Finally, three Y chromosome-specific fragments (Contig-275834, Contig-359642, and Contig-418354) were identified, and specific primers were obtained to distinguish the sex of snakehead. Additionally, a pair of primers of Contig-275834 (275834X/Y-F and 275834X/Y-R) was exploited to distinguish XX females, XY males, and YY super-males, whose amplification products of different lengths were produced for different sexes. Therefore, our work demonstrated the ability of NGS data in identification of sex-specific markers, and the pipeline adopted in our study could be applied in any species of sex differentiation. Furthermore, the sex-specific markers have tremendous potential for improving the efficiency of all-male breeding practices in snakehead.

13.
Gigascience ; 6(4): 1-5, 2017 04 01.
Artículo en Inglés | MEDLINE | ID: mdl-28327946

RESUMEN

The Northern snakehead (Channa argus), a member of the Channidae family of the Perciformes, is an economically important freshwater fish native to East Asia. In North America, it has become notorious as an intentionally released invasive species. Its ability to breathe air with gills and migrate short distances over land makes it a good model for bimodal breath research. Therefore, recent research has focused on the identification of relevant candidate genes. Here, we performed whole genome sequencing of C. argus to construct its draft genome, aiming to offer useful information for further functional studies and identification of target genes related to its unusual facultative air breathing. Findings: We assembled the C. argus genome with a total of 140.3 Gb of raw reads, which were sequenced using the Illumina HiSeq2000 platform. The final draft genome assembly was approximately 615.3 Mb, with a contig N50 of 81.4 kb and scaffold N50 of 4.5 Mb. The identified repeat sequences account for 18.9% of the whole genome. The 19 877 protein-coding genes were predicted from the genome assembly, with an average of 10.5 exons per gene. Conclusion: We generated a high-quality draft genome of C. argus, which will provide a valuable genetic resource for further biomedical investigations of this economically important teleost fish.


Asunto(s)
Genoma , Genómica , Perciformes/genética , Animales , Biología Computacional/métodos , Tamaño del Genoma , Genómica/métodos , Anotación de Secuencia Molecular , Familia de Multigenes , Perciformes/clasificación , Filogenia , Secuencias Repetitivas de Ácidos Nucleicos , Análisis de Secuencia de ADN , Secuenciación Completa del Genoma
14.
Fish Physiol Biochem ; 41(6): 1419-33, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-26159319

RESUMEN

Channa argus is one of the most commercially important fish species in China. Studies show that males of C. argus grow faster than females at the same age. In order to explore the sex differentiation mechanism of C. argus, we isolated the full length of the sex-related gene Foxl2 cDNA and analysed its expression patterns during gonadal sex differentiation. Alignment of known Foxl2 amino acid sequences from vertebrates confirmed the conservation of the Foxl2 open reading frame, especially the forkhead domain and C-terminal region. Quantitative RT-PCR revealed that Foxl2 is predominantly expressed in brain, pituitary, gill and ovary, with its highest level in ovary but low levels in testis and other tissues, reflecting a potential role for Foxl2 in the brain-pituitary-gonad axis in C. argus. Our ontogenetic stage data showed that C. argus Foxl2 expression was significantly upregulated from 1 to 11 days posthatching (dph) and that the initiation of expression preceded the first anatomical ovarian differentiation (27 dph), suggesting that Foxl2 might play a potential role in early gonadal sex differentiation in C. argus. In addition, the Foxl2 protein was primarily located in granulosa cells surrounding the oocytes of mature C. argus, implying that Foxl2 may have a basic function in granulosa cell differentiation and the maintenance of oocytes.


Asunto(s)
Proteínas de Peces/metabolismo , Factores de Transcripción Forkhead/metabolismo , Gónadas/fisiología , Perciformes/fisiología , Diferenciación Sexual/genética , Secuencia de Aminoácidos , Animales , Clonación Molecular , Femenino , Proteínas de Peces/genética , Factores de Transcripción Forkhead/genética , Regulación del Desarrollo de la Expresión Génica , Células de la Granulosa/fisiología , Masculino , Datos de Secuencia Molecular , Ovario/fisiología , Perciformes/genética , Testículo/fisiología
15.
J Biochem Mol Toxicol ; 28(7): 292-301, 2014 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-24891093

RESUMEN

The voltage-dependent anion channel (VDAC) is a highly conserved integral protein of mitochondria in different eukaryotic species. It forms a selective channel in the mitochondrial outer membrane that serves as the controlled pathway for small metabolites and ions. In this study, a VDAC gene, EcVDAC1, was isolated from orange-spotted grouper (Epinephelus coioides). The EcVDAC1 exhibits ubiquitous expression in various tissues of orange-spotted grouper and is upregulated in liver, gill, and spleen after stimulation with lipopolysaccharides (LPS). Subcellular localization analysis shows that the EcVDAC1 protein colocalized with the mitochondria. A caspase-3 assay demonstrates that overexpression of the EcVDAC1 induced apoptotic cell death in fathead minnow cells. The data presented in this study provide new information regarding the relationship between LPS and the EcVDAC1 gene, suggesting that the fish VDAC1 gene may play an important role in antibacterial immune response.


Asunto(s)
Peces/metabolismo , Canal Aniónico 1 Dependiente del Voltaje/metabolismo , Secuencia de Aminoácidos , Animales , Apoptosis , Línea Celular , China , Regulación de la Expresión Génica , Datos de Secuencia Molecular , Filogenia , Fracciones Subcelulares/metabolismo , Transfección , Canal Aniónico 1 Dependiente del Voltaje/química , Canal Aniónico 1 Dependiente del Voltaje/genética
16.
Mitochondrial DNA ; 24(4): 362-4, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23379357

RESUMEN

This paper reports the complete mitochondrial genome (mitogenome) sequence of Channa maculate (Taiwan snakehead). The mitogenome of Taiwan snakehead was determined to be 16,558 bp in length. It contains 13 typical vertebrate protein-coding genes, 22 tRNA genes, 2 rRNA genes, 1 OL (putative light-strand origin of replication), and 1 putative control region (D-loop). This is the first report on the complete mitogenome sequence of Channa maculata.


Asunto(s)
ADN Mitocondrial/genética , Genes Mitocondriales/genética , Genoma Mitocondrial/genética , Perciformes/genética , Animales , Composición de Base , Secuencia de Bases , Orden Génico/genética , Tamaño del Genoma/genética , Datos de Secuencia Molecular , Oligonucleótidos/genética , Análisis de Secuencia de ADN , Taiwán
17.
Food Chem ; 136(2): 682-8, 2013 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-23122114

RESUMEN

The Pearl River Delta (PRD) is located in the Southern part of China and is the main region for fish culture in Guangdong Province. In order to assess the potential health risks associated with dietary consumption of mercury, hair samples from 91 urban, town and fishing village residents, 37 species of fish, cereal, vegetables, and meat samples were collected. The average total mercury (THg) and methylmercury (MeHg) concentrations in hair were 1.08 ± 0.94 and 0.58 ± 0.59 µg/g, respectively. Daily Hg intake via fish consumption is significantly correlated with THg and MeHg accumulated in human hair (r=0.48, p<0.01; r=0.43, p<0.01). The estimated daily intake of Hg via different food types showed that both fish and cereal consumption were the two main routes of Hg exposure for residents in the sampling areas. Besides food intake, smoking was also an important source for daily THg intake in the smoke group, contributing 11-18% to EDI of THg.


Asunto(s)
Contaminación de Alimentos/análisis , Cabello/química , Mercurio/análisis , Adulto , Animales , China , Productos Agrícolas/química , Ingestión de Alimentos , Monitoreo del Ambiente , Femenino , Peces , Análisis de los Alimentos , Cabello/metabolismo , Humanos , Masculino , Carne/análisis , Mercurio/metabolismo , Compuestos de Metilmercurio/análisis , Compuestos de Metilmercurio/metabolismo , Persona de Mediana Edad , Alimentos Marinos/análisis , Verduras/química , Adulto Joven
18.
Fish Shellfish Immunol ; 34(1): 273-9, 2013 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-23178692

RESUMEN

GRIM-19 is a nuclear encoded subunit of complex I that has been implicated in apoptosis. The protein participates in multiple functions including the innate immune response. GRIM-19 has been studied in humans and other mammals; however, fish GRIM-19 has not been well characterized. In this study, a new GRIM-19 gene, EcGRIM-19, was isolated from the orange-spotted grouper (Epinephelus coioides) cDNA library, which was constructed following LPS treatment. EcGRIM-19 is a 582-bp gene that encodes a 144-amino acid protein. The gene is a true ortholog of mammalian GRIM-19. EcGRIM-19 exhibits ubiquitous and constitutive expression in the different tissues of the orange-spotted grouper. The expression levels of EcGRIM-19 are altered in the gill, spleen, kidney and liver after induction with LPS. The subcellular localization analysis demonstrated that the EcGRIM-19 protein is localized predominantly in the mitochondria. In addition, amino acids 30-50 of the protein are responsible for the mitochondrial localization of EcGRIM-19. The caspase assay demonstrated that the overexpression of GRIM-19 enhanced the cellular sensitivity to interferon(IFN)-ß- and retinoic acid (RA)-induced death in HeLa cells. The data presented in this study are important for further understanding the EcGRIM-19 gene function in fish.


Asunto(s)
Lubina/genética , Lubina/metabolismo , Proteínas de Peces/genética , Proteínas de Peces/metabolismo , Secuencia de Aminoácidos , Animales , Apoptosis , Caspasas/metabolismo , Clonación Molecular , ADN Complementario/genética , Células HeLa , Humanos , Interferón beta/metabolismo , Lipopolisacáridos/farmacología , Datos de Secuencia Molecular , Especificidad de Órganos , Filogenia , ARN/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Alineación de Secuencia , Transfección , Tretinoina/metabolismo
19.
Environ Sci Pollut Res Int ; 20(7): 4484-95, 2013 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-23247527

RESUMEN

This study investigated the extent of arsenic (As) contamination in five common species of freshwater fish (northern snakehead [Channa argus], mandrarin fish [Siniperca chuatsi], largemouth bass [Lepomis macrochirous], bighead carp [Aristichthys nobilis] and grass carp [Ctenopharyngodon idellus]) and their associated fish pond sediments collected from 18 freshwater fish ponds around the Pearl River Delta (PRD). The total As concentrations detected in fish muscle and sediment in freshwater ponds around the PRD were 0.05-3.01 mg kg(-1) wet weight (w. wt) and 8.41-22.76 mg kg(-1) dry weight (d. wt), respectively. In addition, the As content was positively correlated (p < 0.05) to total organic carbon (TOC) contents in sediments. Biota sediment accumulation factor (BSAF) showed that omnivorous fish and zooplankton accumulated higher concentrations of heavy metals from the sediment than carnivorous fish. In addition, feeding habits of fish also influence As accumulation in different fish species. In this study, two typical food chains of the aquaculture ponds were selected for investigation: (1) omnivorous food chain (zooplankton, grass carp and bighead carp) and (2) predatory food chain (zooplankton, mud carp and mandarin fish). Significant linear relationships were obtained between log As and δ (15)N. The slope of the regression (-0.066 and -0.078) of the log transformed As concentrations and δ (15)N values, as biomagnifications power, indicated there was no magnification or diminution of As from lower trophic levels (zooplankton) to fish in the aquaculture ponds. Consumption of largemouth bass, northern snakehead and bighead carp might impose health risks of Hong Kong residents consuming these fish to the local population, due to the fact that its cancer risk (CR) value exceeded the upper limit of the acceptable risk levels (10(-4)) stipulated by the USEPA.


Asunto(s)
Arsénico/análisis , Monitoreo del Ambiente/métodos , Estanques/análisis , Ríos/química , Contaminantes Químicos del Agua/análisis , Animales , Acuicultura , Peces , Cadena Alimentaria , Sedimentos Geológicos/análisis , Hong Kong , Metales Pesados/análisis , Estanques/química , Control de Calidad , Medición de Riesgo , Zooplancton/efectos de los fármacos , Zooplancton/crecimiento & desarrollo
20.
Huan Jing Ke Xue ; 33(6): 1808-15, 2012 Jun.
Artículo en Chino | MEDLINE | ID: mdl-22946159

RESUMEN

In order to investigate the heavy metal concentrations and their potential ecological risks in surface sediments of lower reaches and estuary of Pearl River, 21 bottom sediment samples were collected from lower reaches and estuary of Pearl River. Total contents of Cr, Mn, Fe, Co, Ni, Cu, Zn, As, Se, Cd, Sb, Pb and Hg in these samples were measured by the inductively coupled plasma mass spectrometry (ICP-MS) and the atomic fluorescence spectrometry (AFS) and using the index of geoaccumulation and the potential ecological risk index to evaluate the pollution degree of heavy metals in the sediments. Results indicated that the concentration of total Fe and total Mn were 41658.73 and 1104.73 mg x kg(-1) respectively and toxic trace metals, such as Cr, Co, Ni, Cu, Zn, As, Se, Cd, Sb, Pb and Hg were 86.62, 18.18, 54.10, 80.20, 543.60, 119.55, 4.28, 10.60, 20.26, 104.58 and 0.520 mg x kg(-1). The descending order of pollution degree of various metals is: Cd > As approximately Zn > Hg > Pb approximately Cu approximately Cr, while the single potential ecological risk followed the order: Cd > Hg > As > Cu > Pb > Zn > Cr. The pollution extent and potential ecological risk of Cd were the most serious among all heavy metals. The distribution pattern of Cd individual potential ecological risk indices is exactly the same as that of general potential ecological risk indices for all heavy metals. Clustering analysis indicates that the sampling stations may be classified into five groups which basically reflected the characteristics of the heavy metal contamination and sedimentation environments along the different river reaches in lower reaches and estuary of Pearl Rive. In general, the serious heavy metal pollution and the high potential ecological risk existed in three river reaches: Chengcun-Shawan, Chengcun-Shundegang and Waihai-Hutiaomen. The pollution degree and potential ecological risk are higher in related river reaches of Beijiang than that in other lower reaches and estuary of Pearl River.


Asunto(s)
Sedimentos Geológicos/análisis , Metales Pesados/análisis , Contaminantes Químicos del Agua/análisis , China , Análisis por Conglomerados , Fenómenos Ecológicos y Ambientales , Monitoreo del Ambiente , Medición de Riesgo , Ríos
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