- Deshou Wang currently works at the School of Life Sciences, Southwest University in China. He does research in Molecu... moreDeshou Wang currently works at the School of Life Sciences, Southwest University in China. He does research in Molecular Biology, Genetics and Endocrinology. Their current projects are 'Sexual plasticity and sex control in fish' , 'Body color fromation in tilapia' and 'Creation of tilapia new germplasm for aquaculture'.edit
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Research Interests: Biology, CRISPR, CAS, Gene, Green Fluorescent Protein, and 3 morePlasmid, Genome Editing, and Reporter gene
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Argonaute proteins are key components of the small RNA-induced silencing complex and have multiple roles in RNA-directed regulatory pathways. Argonaute genes can be divided into two subfamilies: the Ago (interacting with microRNA/small... more
Argonaute proteins are key components of the small RNA-induced silencing complex and have multiple roles in RNA-directed regulatory pathways. Argonaute genes can be divided into two subfamilies: the Ago (interacting with microRNA/small interfering RNA) and Piwi subfamilies (interacting with piwi-interacting RNAs (piRNAs)). In the present study, genome-wide analyses firstly yielded the identification of different members of Agos and Piwis in the tilapia, coelacanth, spotted gar, and elephant shark. The additional teleost Ago3b was generated following the fish-specific genome duplication event. Selective pressure analysis on Agos and Piwis between cichlids and other teleosts showed an accelerated evolution of Piwil1 in the cichlid lineages, and the positive selected sites were located in the region of PIWI domain, suggesting that these amino acid substitutions are adapt to targeted cleavage of messenger RNA (mRNA) in cichlids. Ago1 and Ago4 were detected at higher levels at 5 days aft...
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The furin-domain-containing peptide R-spondin 1 (RSPO1) has recently emerged as an important regulator of ovarian development, upregulating the WNT/β-catenin pathway to oppose testis formation in mammals. However, little information has... more
The furin-domain-containing peptide R-spondin 1 (RSPO1) has recently emerged as an important regulator of ovarian development, upregulating the WNT/β-catenin pathway to oppose testis formation in mammals. However, little information has been reported on the Rspo1 signaling pathway in teleosts. In this study, Rspo1 was isolated from the gonads of the Nile tilapia, Oreochromis niloticus. An in situ hybridization analysis demonstrated that Rspo1 is expressed in the germ cells of the ovary and the testis. An ontogenic analysis demonstrated that Rspo1 expression is upregulated just before meiotic initiation in both the ovary and testis during the early developmental stages of the tilapia. The expression pattern is sexually dimorphic from 20 days after hatching, with higher expression in the ovary. The reduction of Rspo1 expression by transcription activator-like (TAL) effector nuclease (TALEN) caused retarded ovarian development, the ectopic expression of male-dominant genes, and increas...
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Two isoforms of Vasa cDNA, derived from the 5′alternative splicing of the same gene, were isolated and characterized in Southern catfish, Silurus meridionalis, by reverse transcription polymerase chain reaction (RT-PCR) and rapid... more
Two isoforms of Vasa cDNA, derived from the 5′alternative splicing of the same gene, were isolated and characterized in Southern catfish, Silurus meridionalis, by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. Analysis of the nucleotide sequences revealed that the full length cDNA of Southern catfish Vasa (scVasa) comprises 2525 base pairs (bp) with an open reading frame (ORF) of 1989 bp, encoding 662 amino acids, while that of Vasa short form (scVasa-s) comprises 2438 bp with an ORF of 1926 bp, encoding 641 amino acids. Except the difference in 5′-untranslated region, scVasa-s also lacks a part of the 5′-ORF region found in the scVasa. Both of the two deduced amino acid sequences contain all the eight conserved motifs characteristic to the DEAD-box protein family and four arginine-glycine-glycine motifs unique to Vasa proteins. scVasa showed the highest similarity to Vasa homolog of giebel carp (73.3%). Tissue distribut...
We report the generation of a chimeric monoclonal antibody (ch806) with specificity for an epitope on the epidermal growth factor receptor (EGFR) that is different from that targeted by all other anti-EGFR therapies. Ch806 antibody is... more
We report the generation of a chimeric monoclonal antibody (ch806) with specificity for an epitope on the epidermal growth factor receptor (EGFR) that is different from that targeted by all other anti-EGFR therapies. Ch806 antibody is reactive to both de2-7 and overexpressed wild-type (wt) EGFR but not native EGFR expressed in normal tissues at physiological levels. Ch806 was stably expressed in CHO (DHFR -/-) cells and purified for subsequent characterisation and validated for use in preliminary immunotherapy investigations. Ch806 retained the antigen binding specificity and affinity of the murine parental antibody. Furthermore, ch806 displayed enhanced antibody-dependent cellular cytotoxicity against target cells expressing the 806 antigen in the presence of human effector cells. Ch806 was successfully radiolabelled with both iodine-125 and indium-111 without loss of antigen binding affinity or specificity. The radioimmunoconjugates were stable in the presence of human serum at 37...
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Genomic DNA prepared from the Nile tilapia testis was used for Fosmid library construction. It was randomly sheared to-40 kb fragments. End-repair and recovery of the size-fractionated DNA were manipulated according to the... more
Genomic DNA prepared from the Nile tilapia testis was used for Fosmid library construction. It was randomly sheared to-40 kb fragments. End-repair and recovery of the size-fractionated DNA were manipulated according to the manufacturer's protocols. Ligation of ...
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From June to July 1988, larvae of Mystus macropterus (Bleeker) were obtained by artificial propagation of spawners collected from the Jialing River, China. Larvae grown in water temperatures ranging from 26 to 29°C were fully developed at... more
From June to July 1988, larvae of Mystus macropterus (Bleeker) were obtained by artificial propagation of spawners collected from the Jialing River, China. Larvae grown in water temperatures ranging from 26 to 29°C were fully developed at approximately 20 days. The newly‐hatched larvae measured 6.0–7.5 mm t.l., exceeding the dimensions of any known newly‐hatched larvae of freshwater catfishes indigenous to China. Five days after hatching, when the larvae were 11.4 mm t.l. and the prolarval stage was complete, exogenous feeding commenced. Twelve days after hatching, at 18.4 mm t.l., the yolk sac disappeared, organogenesis was almost complete and the juvenile period began. At 20 days post‐hatching, at 23.0 mm t.l., the lateral line system had formed and the juveniles resembled the adults with respect to all external features, which signalled the end of thejuvenile period. Comparison of the larval development in eight species ofeconomic freshwater catfishes shows that the most signific...
Research Interests: Zoology, Biology, Ecology, Fish Biology, Fisheries Sciences, and 6 moreFishery, Larva, Juvenile, Larval Development, Hatching, and yolk sac
Research Interests: Zoology, Biology, Ecology, Medicine, Cell line, and 13 moreFemale, Animals, Karyotyping, Fisheries Sciences, Catfish, Karyotype, Cell Proliferation, Ovary, Somatic Cell Count, Fish Physiology and Biochemistry, Granulosa Cells, Gene expression profiling, and reverse transcriptase polymerase chain reaction
Research Interests: Genetics, Biology, Medicine, Gene expression, In Situ Hybridization, and 15 morecyclic AMP, Female, Animals, Male, Molecular Phylogenetics and Evolution, Cattle, Molecular cloning, Amino Acid Sequence, Base Sequence, Ovary, Biochemistry and cell biology, Molecular Sequence Data, Oncorhynchus, Medical biochemistry and metabolomics, and Gonadotropin
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Research Interests: Biology, Medicine, Gene expression, In Situ Hybridization, real time PCR, and 15 moreCortisol, Kidney, Animals, Biochemical, Gene, Oocyte Maturation, Molecular cloning, Time Factors, Quntitative Thin Layer Chromatography, Enzymatic Activity, Biochemistry and cell biology, Gene Expression Regulation, teleost fish, hydrocortisone, and Medical biochemistry and metabolomics
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Premelanosome protein (pmel) is a key gene for melanogenesis. Mutations in this gene are responsible for white plumage in chicken, but its role in pigmentation of fish remains to be demonstrated. In this study, we found that most fishes... more
Premelanosome protein (pmel) is a key gene for melanogenesis. Mutations in this gene are responsible for white plumage in chicken, but its role in pigmentation of fish remains to be demonstrated. In this study, we found that most fishes have 2 pmel genes arising from the teleost-specific whole-genome duplication. Both pmela and pmelb were expressed at high levels in the eyes and skin of Nile tilapia. We mutated both genes in tilapia using CRISPR/Cas9. Homozygous mutation of pmela resulted in yellowish body color with weak vertical bars and a hypopigmented retinal pigment epithelium (RPE) due to significantly reduced number and size of melanophores. In contrast, we observed an increased number and size of xanthophores in mutants compared to wild-type fish. Homozygous mutation of pmelb resulted in a similar, but milder phenotype than pmela−/− mutants. Double mutation of pmela and pmelb resulted in loss of additional melanophores compared to the pmela−/− mutants, and also an increase i...
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Table S1. Result of tblastn search against teleost genomes (XLSX 223Â kb)
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Figure S1. Phylogenetic trees of all Teratorn-like virus copies obtained by blast search Neighbor-joining trees of each herpesvirus gene are shown. Kimuraâ s two-parameter model, assuming uniform evolutionary rates among sites, was used... more
Figure S1. Phylogenetic trees of all Teratorn-like virus copies obtained by blast search Neighbor-joining trees of each herpesvirus gene are shown. Kimuraâ s two-parameter model, assuming uniform evolutionary rates among sites, was used as nucleotide substitution model. For DNA polymerase, major capsid protein and membrane glycoprotein, phylogenetic trees were also constructed from the first and second half of the genes, since some sequences contain only a part of the coding region. For terminase, phylogenetic trees were independently constructed for each of the three exons. Numbers above the trees indicate the corresponding regions relative to the CDS of subtype 1 medaka Teratorn. Sequences marked by magenta were used for phylogenetic trees in Fig. 1b and Fig. 4b, those marked in orange were used for phylogenetic trees in Fig. S2 and S3, and those marked with cyan are same as those in Fig. 2. The bars represent the number of substitutions per site. (PDF 234Â kb)
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Table S6. Parameters used for phylogenetic analyses (XLSX 34Â kb)
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The involvement of sex steroids in sex determination and differentiation is relatively conserved among non-mammalian vertebrates, especially in fish. Thanks to the advances in genome sequencing and genome editing, significant progresses... more
The involvement of sex steroids in sex determination and differentiation is relatively conserved among non-mammalian vertebrates, especially in fish. Thanks to the advances in genome sequencing and genome editing, significant progresses have been made in the understanding of steroidogenic pathway and hormonal regulation of sex determination and differentiation in fish. It seems that loss of function study of single gene challenges the traditional views that estrogen might not be required for ovarian differentiation and androgen might not be needed for testicular development, but it is not so in essence. Steroidogenic enzymes can be classified into two categories based on expression and enzyme activities in fish. One type, encoded by star2, cyp17a1 and cyp19a1a, is involved in estrogen production and exclusively expressed in the gonads. Mutation of these genes results in the up-regulation of male pathway genes and sex reversal from genetic female to male. The other type, encoded by the duplicated paralogs of the above genes, including star1, cyp11a1, cyp17a2 and cyp19a1b, as well as cyp11c1 gene, is dominantly expressed both in gonads and extra-gonadal tissues. Mutation of these genes alters the steroids (androgen, DHP and cortisol) production and spermatogenesis, fertility, secondary sexual characteristics and sexual behavior, but usually does not affect the sex differentiation. For the estrogen receptors (esr1, esr2a and esr2b), single mutation failed to, but double and triple mutation leads to sex reversal from female to male, indicating that at least two estrogen receptors might be required to mediate the role of estrogen in sex determination proved by gene editing experiments. Taken together, results from gene editing enrich our understanding of steroid synthesis pathways and further confirm the critical role of estrogen in female sex determination by antagonizing the male pathway in fish.
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The diverse color patterns of cichlid fishes play an important role in mate choice and speciation. Here we develop the Nile tilapia (Oreochromis niloticus) as a model system for studying the developmental genetics of cichlid color... more
The diverse color patterns of cichlid fishes play an important role in mate choice and speciation. Here we develop the Nile tilapia (Oreochromis niloticus) as a model system for studying the developmental genetics of cichlid color patterns. We identified 4 types of pigment cells: melanophores, xanthophores, iridophores and erythrophores, and characterized their first appearance in wild-type fish. We mutated 25 genes involved in melanogenesis, pteridine metabolism, and the carotenoid absorption and cleavage pathways. Among the 25 mutated genes, 13 genes had a phenotype in both the F0 and F2 generations. None of F1 heterozygotes had phenotype. By comparing the color pattern of our mutants with that of red tilapia (Oreochromis spp), a natural mutant produced during hybridization of tilapia species, we found that the pigmentation of the body and eye is controlled by different genes. Previously studied genes like mitf, kita/kitlga, pmel, tyrb, hps4, gch2, csf1ra, pax7b, and bco2b were pr...
Research Interests: Genetics, Biology, Medicine, Heredity, Chromatophore, and 4 moreGene, Nile Tilapia, Tilapia, and Oreochromis
Chromosome size and morphology vary within and among species, but little is known about either the proximate or ultimate causes of these differences. Cichlid fish species in the tribe Oreochromini share an unusual megachromosome that is... more
Chromosome size and morphology vary within and among species, but little is known about either the proximate or ultimate causes of these differences. Cichlid fish species in the tribe Oreochromini share an unusual megachromosome that is ~3 times longer than any of the other chromosomes. This megachromosome functions as a sex chromosome in some of these species. We explore two hypotheses of how this sex megachromosome may have evolved. The first hypothesis proposes that it developed by the accumulation of repetitive elements as recombination was reduced around a dominant sex-determination locus, as suggested by traditional models of sex chromosome evolution. An alternative hypothesis is that the megachromosome originated via the fusion of an autosome with a highly-repetitive B chromosome, one of which had carried a sex-determination locus. Here we test these hypotheses using comparative analysis of several chromosome-scale cichlid and teleost genomes. We find the megachromosome consi...
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The origin and evolution of molecular mechanisms underlying the self-renewal and differentiation of spermatogonial stem cells (SSCs) are fundamental questions in stem cell biology as well as reproduction medicine. In mammals, glial cell... more
The origin and evolution of molecular mechanisms underlying the self-renewal and differentiation of spermatogonial stem cells (SSCs) are fundamental questions in stem cell biology as well as reproduction medicine. In mammals, glial cell line-derived neurotrophic factor (GDNF) is crucial for SSC self-renewal and maintenance. However, in nonmammals, the role of Gdnf in SSCs still remains unknown. In this study, we report that the two GDNF homologs from medaka fish (Oryzias latipes), namely OlGdnfa and OlGdnfb, can promote proliferation activity and retain the spermatogonial property of SG3, a spermatogonial cell line derived from adult medaka showing the intrinsic property of SSCs by self-renewal and differentiation potential during 2 years of culture. Cloning and sequencing led to the identification of two cDNA sequences as Olgdnfa and Olgdnfb, which are 780-nt and 744-nt in length for 253 and 245 amino acid residues, respectively. Both are homologs of mammalian GDNF and share over 4...
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Two β-catenin (β-cat) genes exist in teleosts but little is known about their expression and function in ovarian development. We identified β-cat1 and β-cat2 from the Nile tilapia. β-cat1 and β-cat2 displayed a similar expression pattern... more
Two β-catenin (β-cat) genes exist in teleosts but little is known about their expression and function in ovarian development. We identified β-cat1 and β-cat2 from the Nile tilapia. β-cat1 and β-cat2 displayed a similar expression pattern in the ovary during development, and were mainly expressed in the oogonia and oocytes. In luciferase assays, β-cat1 activated the TOPFlash reporter dose-dependently, whereas β-cat2 failed to do so. Cotransfection of β-cat1 and β-cat2 synergistically enhanced the expression of the reporter. A specific interaction between β-cat1 and β-cat2 was also observed in a mammalian two-hybrid assay. Furthermore, tilapia recombinant Dkk1, an inhibitor of the β-cat pathway, decreased β-cat1 and β-cat2, while increased sox9, dmrt1, cyp11b2 and foxl2 expression in the in vitro cultured tilapia ovary, which could be abolished by simultaneous treatment with Bio, an agonist of β-cat. Consistently, β-cat1 or β-cat2 knockdown in XX fish by TALENs caused the retardation ...
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The Sox transcription factor family is characterized with the presence of a Sry-related high-mobility group (HMG) box and plays important roles in various biological processes in animals, including sex determination and differentiation,... more
The Sox transcription factor family is characterized with the presence of a Sry-related high-mobility group (HMG) box and plays important roles in various biological processes in animals, including sex determination and differentiation, and the development of multiple organs. In this study, 27 Sox genes were identified in the genome of the Nile tilapia (Oreochromis niloticus), and were classified into seven groups. The members of each group of the tilapia Sox genes exhibited a relatively conserved exon-intron structure. Comparative analysis showed that the Sox gene family has undergone an expansion in tilapia and other teleost fishes following their whole genome duplication, and group K only exists in teleosts. Transcriptome-based analysis demonstrated that most of the tilapia Sox genes presented stage-specific and/or sex-dimorphic expressions during gonadal development, and six of the group B Sox genes were specifically expressed in the adult brain. Our results provide a better und...
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To address the roles of doublesex and mab-3-related transcription factor 1 (Dmrt1), forkhead transcription factor gene 2 (Foxl2), and aromatase in sex differentiation of Southern catfish, the cDNA sequences of these genes were isolated... more
To address the roles of doublesex and mab-3-related transcription factor 1 (Dmrt1), forkhead transcription factor gene 2 (Foxl2), and aromatase in sex differentiation of Southern catfish, the cDNA sequences of these genes were isolated from the gonads. Dmrt1a and Dmrt1b were found to be expressed in the gonads, being higher in the testis. A low expression level of Dmrt1b was also detected in the intestine and kidney of the male. Foxl2 was found to be expressed extensively in the brain (B), pituitary (P), gill and gonads (G), with the highest level in the ovary, indicating the possible involvement of Foxl2 in the B–P–G axis. Cytochrome P450 (Cyp)19b was found to be expressed in the brain, spleen, and gonads, while Cyp19a was only expressed in the gonads and spleen. All-female Southern catfish fry were treated with fadrozole (F), tamoxifen (TAM), and 17β-estradiol (E2) respectively, from 5 to 25 days after hatching (dah). The expression levels of these genes were measured at 65 dah. I...
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Previous study from our laboratory on thiourea-induced thyroid hormone depletion in mature male catfish demonstrated that thyroid hormones play a significant role in testicular function. In the present study, we aimed to analyze the... more
Previous study from our laboratory on thiourea-induced thyroid hormone depletion in mature male catfish demonstrated that thyroid hormones play a significant role in testicular function. In the present study, we aimed to analyze the changes in the expression pattern of several steroidogenic enzyme genes after thyroid hormone depletion using semi quantitative RT-PCR in both adult male and female catfish. There was a marked decrease in the 11beta-hydroxylase expression in the testis and liver while no change was observed in case of kidney. A significant decrease in 11beta-hydroxysteroid dehydrogenase transcript level in testis, liver and kidney were observed in the thiourea treated males. The results obtained corroborated with our earlier findings of testicular regression after thyroid hormone depletion. In females, expression of aromatase transcript increased in experimental group compared to control. There was no considerable change observed in the transcript level of 3beta-hydroxysteroid dehydrogenase, P450 17alpha-hydroxylase/C17-20-lyase, and 20beta-hydroxysteroid dehydrogenase in both males and females. Thus, thyroid hormones might exert modulating effect on steroidogenic enzyme genes at the transcription level.
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The objective of the present study is to observe the effect of exogenous steroids, methyl testosterone (MT) and ethynyl estradiol (EEL) on gonadal differentiation and analyze its effect on the expression of several genes during testicular... more
The objective of the present study is to observe the effect of exogenous steroids, methyl testosterone (MT) and ethynyl estradiol (EEL) on gonadal differentiation and analyze its effect on the expression of several genes during testicular and ovarian differentiation in juvenile catfish. Exogenous hormone treatments (MT and EEL) were given by immersion at different days of hatching. The histological analysis revealed that the EEL- and MT-treatments resulted in the initiation of ovarian and testicular differentiation, respectively. This is further supported by specific expression of two forms of DMRT1 in the MT-treated group but not in the EEL-treated group at 47 days after hatching (dah). The reverse is true for the expression of ovarian aromatase. Results of the semi-quantitative RT-PCR show that brain aromatase transcript levels are high in 47 dah control (histologically female) and 47 dah EEL-treated fish, as compared to 47 dah MT-treated fish. At 60 dah, brain aromatase showed elevation in its expression. Interestingly, the expression pattern of 3 beta-HSD did not show any change in EEL- and MT-treated fish. The present study also provides a strategy to study sex differentiation, for those species where genetic sex population is unavailable.
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Research Interests: Endocrinology, Biology, Breeding, Ecology, Environmental Change, and 15 moreEstrogen Receptor, Internal Medicine, Female, Animals, Disorders of sex development, Feminization, Early development, Fisheries Sciences, Catfish, Estrogens, Artificial insemination, Aromatase Inhibitor, Gonads, aromatase, and Expression pattern
Research Interests: Molecular Biology, Biology, Ecology, Medicine, Phylogeny, and 15 moreAnimals, Gene Order, Phylogenetic analysis, Fisheries Sciences, Freshwater Fish, Pituitary Gland, Amino Acid Sequence, Base Sequence, Cyprinidae, Minnow, Gene Expression Regulation, Gene expression profiling, Common Carp, Molecular Sequence Data, and cDNA cloning
In teleost fish, sex steroids are involved in sex determination, sex differentiation, and fertility. Cyp17a1 (Cytochrome P450 family 17 subfamily A member 1) is thought to play essential roles in fish steroidogenesis. Therefore, to... more
In teleost fish, sex steroids are involved in sex determination, sex differentiation, and fertility. Cyp17a1 (Cytochrome P450 family 17 subfamily A member 1) is thought to play essential roles in fish steroidogenesis. Therefore, to further understand its roles in steroidogenesis, sex determination, and fertility in fish, we constructed a cyp17a1 gene mutant in Nile tilapia (Oreochromis niloticus). In XX fish, mutation of the cyp17a1 gene led to a female-to-male sex reversal with a significant decline in 17β-estradiol (E2) and testosterone (T) production, and ectopic expression of male-biased markers (Dmrt1 and Gsdf) in gonads from the critical window of sex determination. Sex reversal was successfully rescued via T or E2 administration, and ovarian characteristics were maintained after termination of E2 supplementation in the absence of endogenous estrogen production in cyp17a1–/– XX fish. Likewise, deficiencies in T and 11-ketotestosterone (11-KT) production in both cyp17a1–/– XX s...
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Text S2. Trimmed alignments used for phylogenetic tree construction (TXT 3590Â kb)
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Spermatogenesis is a complex process in which spermatogonial stem cells differentiate and develop into mature spermatozoa. The transcriptional regulatory network involved in fish spermatogenesis remains poorly understood. Here, we... more
Spermatogenesis is a complex process in which spermatogonial stem cells differentiate and develop into mature spermatozoa. The transcriptional regulatory network involved in fish spermatogenesis remains poorly understood. Here, we demonstrate in Nile tilapia that the Sox transcription factor family member Sox30 is specifically expressed in the testes and mainly localizes to spermatocytes and spermatids. CRISPR/Cas9-mediated sox30 mutation results in abnormal spermiogenesis, reduction of sperm motility, and male subfertility. Comparative transcriptome analysis shows that sox30 mutation alters the expression of genes involved in spermatogenesis. Further chromatin immunoprecipitation followed by high-throughput sequencing (ChIP-seq), ChIP-PCR, and luciferase reporter assays reveal that Sox30 positively regulates the transcription of ift140 and ptprb, two genes involved in spermiogenesis, by directly binding to their promoters. Taken together, our data indicate that Sox30 plays essential roles in Nile tilapia spermatogenesis by directly regulating the transcription of the spermiogenesis-related genes ift140 and ptprb.
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The Mozambique tilapia (Oreochromis mossambicus) is a fascinating taxon for evolutionary and ecological research. It is an important food fish and one of the most widely distributed tilapias. Because males grow faster than females,... more
The Mozambique tilapia (Oreochromis mossambicus) is a fascinating taxon for evolutionary and ecological research. It is an important food fish and one of the most widely distributed tilapias. Because males grow faster than females, genetically male tilapia are preferred in aquaculture. However, studies of sex determination and sex control in O. mossambicus have been hindered by the limited characterization of the genome. To address this gap, we assembled a high-quality genome of O. mossambicus, using a combination of high coverage of Illumina and Nanopore reads, coupled with Hi-C and RNA-Seq data. Our genome assembly spans 1,007 Mb with a scaffold N50 of 11.38 Mb. We successfully anchored and oriented 98.6% of the genome on 22 linkage groups (LGs). Based on re-sequencing data for male and female fishes from three families, O. mossambicus segregates both an XY system on LG14 and a ZW system on LG3. The sex-patterned SNPs shared by two XY families narrowed the sex determining regions ...
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Teleosts are important models to study sex chromosomes and sex-determining (SD) genes because they present a variety of sex determination systems. Here, we used Nanopore and Hi-C technologies to generate a high-contiguity chromosome-level... more
Teleosts are important models to study sex chromosomes and sex-determining (SD) genes because they present a variety of sex determination systems. Here, we used Nanopore and Hi-C technologies to generate a high-contiguity chromosome-level genome assembly of a YY southern catfish ( Silurus meridionalis ). The assembly is 750.0 Mb long, with contig N50 of 15.96 Mb and scaffold N50 of 27.22 Mb. We also sequenced and assembled an XY male genome with a size of 727.2 Mb and contig N50 of 13.69 Mb. We identified a candidate SD gene through comparisons to our previous assembly of an XX individual. By resequencing male and female pools, we characterized a 2.38 Mb sex-determining region (SDR) on Chr24. Analysis of read coverage and comparison of the X and Y chromosome sequences showed a Y specific insertion (approx. 500 kb) in the SDR which contained a male-specific duplicate of amhr2 (named amhr2y ). amhr2y and amhr2 shared high-nucleotide identity (81.0%) in the coding region but extremely ...
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The role of glucocorticoids in oogenesis remains to be elucidated. cyp11c1 encodes the key enzyme involved in the synthesis of cortisol, the major glucocorticoid in teleosts. In our previous study, we mutated cyp11c1 in tilapia and... more
The role of glucocorticoids in oogenesis remains to be elucidated. cyp11c1 encodes the key enzyme involved in the synthesis of cortisol, the major glucocorticoid in teleosts. In our previous study, we mutated cyp11c1 in tilapia and analyzed its role in spermatogenesis. In this study, we analyzed its role in oogenesis. cyp11c1+/- XX tilapia showed normal ovarian morphology but poor egg quality, as indicated by the mortality of embryos before 3 d post fertilization, which could be partially rescued by the supplement of exogenous cortisol to the mother fish. Transcriptome analyses revealed reduced expression of maternal genes in the eggs of the cyp11c1+/- XX fish. The cyp11c1-/- females showed impaired vitellogenesis and arrested oogenesis due to significantly decreased serum cortisol. Further analyses revealed decreased serum E2 level and expression of amh, an important regulator of follicular cell development, and increased follicular cell apoptosis in the ovaries of cyp11c1-/- XX fish, which could be rescued by supplement of either exogenous cortisol or E2. Luciferase assays revealed a direct regulation of cortisol and E2 on amh transcription via GRs or ESRs. Taken together, our results demonstrate that cortisol safeguards oogenesis by promoting follicular cell survival probably via Amh signaling.
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Desert hedgehog (DHH) signaling has been reported to be involved in spermatogenesis and the self-renewal of spermatogonial stem cells (SSCs). However, the role of DHH in proliferation of spermatogonia including SSCs remains to be... more
Desert hedgehog (DHH) signaling has been reported to be involved in spermatogenesis and the self-renewal of spermatogonial stem cells (SSCs). However, the role of DHH in proliferation of spermatogonia including SSCs remains to be elucidated. Here, we report that Dhh from medaka (Oryizas latipes) (named as OlDhh) could directly mediate the proliferation of spermatogonia via Smoothened (Smo) signaling. Oldhh is 1362 bp in length and encodes 453 amino acid (aa) residues with more than 50% identity with the homologs in other species. It has expression predominantly restricted to testis. The soluble and tag-free 176-aa mature OlDhh (named as mOlDhh) were successfully obtained by fusing with the N-terminal tag of cleavable 6-histidine and small ubiquitin-related modifier and then removing the tag. Notably, mOlDhh significantly promoted the proliferation of SG3 (a spermatogonial stem cell line from medaka testis) in a dose-dependent manner and spermatogonia in testicular organ culture. Fur...