为初步阐明拟赤梢鱼(Pseudaspius leptocephalus)卵巢发育特征及sox3(sry related high mobility group box 3)基因在其卵巢发育过程中的作用,本研究通过组织切片观察了拟赤梢鱼卵巢早期发育过程,利用RACE技术克隆了该鱼sox3基因c DNA...为初步阐明拟赤梢鱼(Pseudaspius leptocephalus)卵巢发育特征及sox3(sry related high mobility group box 3)基因在其卵巢发育过程中的作用,本研究通过组织切片观察了拟赤梢鱼卵巢早期发育过程,利用RACE技术克隆了该鱼sox3基因c DNA全长序列,并进行生物信息学分析;利用实时荧光定量PCR技术分析了sox3基因在拟赤梢鱼不同组织及性腺不同发育时期的表达模式。结果显示,拟赤梢鱼卵巢在孵化后45 d分化形成,160 d时由Ⅰ期进入Ⅱ期,孵化后360 d,卵巢仍处于Ⅱ期。拟赤梢鱼sox3基因cDNA全长为1800 bp(GenBank登录号:MT952206),编码299个氨基酸,存在保守的HMG(histidine,methionine,glycine-rich)结构域。氨基酸序列比对和系统进化树分析结果显示,拟赤梢鱼SOX3蛋白与斑马鱼(Danio rerio)和翘嘴鲌(Culter alburnus)亲缘关系最近。此外,sox3基因在拟赤梢鱼卵巢中表达量最高,其次是脑和眼睛,在精巢和其他组织中微量表达;在性腺分化过程中,sox3基因在卵巢中表达量显著高于未分化性腺和精巢,在卵巢发育阶段表达量不断升高,而在精巢中持续低水平表达。综上,本研究推测sox3基因主要参与拟赤梢鱼卵巢的分化和发育。展开更多
Evolutionary evidence suggests that Sox3, a member of the high-mobility-group(HMG) family of transcription factors, is an ancestral precursor of Sry and is involved in sex determination similar to Sry. However, there ...Evolutionary evidence suggests that Sox3, a member of the high-mobility-group(HMG) family of transcription factors, is an ancestral precursor of Sry and is involved in sex determination similar to Sry. However, there is limited information regarding the SOX3 gene of the black rockfish(Sebastes schlegeli). In this study, we first isolated SOX3 gene from the gonads of S. schlegeli by homology cloning. The full-length of S. schlegeli SOX3(SsSOX3) c DNA was 1386 bp, comprising a 906-bp open reading frame, which encodes a peptide showing 93.6% and 93.9% homology with the Sox3 proteins of Epinephelus coioides and Oryzias latipe, respectively. Comparison of the cDNA sequence of the Ss SOX3 gene with the corresponding genomic DNA fragment revealed that the SsSOX3 gene consists of a single exon. Phylogenetic analysis demonstrated the evolutionary relationship of Ss SOX3 with other known SOXB1 genes in fish and tetrapods. The promoter region contains binding sites of several transcriptional factors that might participate in the regulation of Ss SOX3 expression. Quantitative real-time PCR analysis indicated that SsSOX3 was expressed in all the investigated larval developmental stages from 1 to 35 days after birth and the level of expression gradually decreased as the development proceeded. SsSOX3 exhibited sexually dimorphic expression in adult gonads, with high expression in the ovary but low expression in the testis. In situ hybridization revealed that SsSOX3 was strongly expressed in oocytes and follicular cells of ovaries but slightly expressed in germ cells of testicular tissues. Therefore, this study suggests that Ss SOX3 plays an important role in oogenesis and ovary differentiation in S. schlegeli.展开更多
In vertebrates, SOX (SRY-related HMG box) genes are thought to be due to major gene duplication events, initially occurring during early stages of metazoan evolution and later during the transition between non-vertebr...In vertebrates, SOX (SRY-related HMG box) genes are thought to be due to major gene duplication events, initially occurring during early stages of metazoan evolution and later during the transition between non-vertebrate chordates and vertebrates. The aim of this study is to examine SOX3 and SOX9 transcription in oogensis and spermatogenesis in a fish model, the blue gourami (Trichogaster trichopterus). In females during oogenesis, SOX9 mRNA levels were lower compared to SOX3 mRNA levels. In males, SOX9 mRNA levels were higher in testes compared to SOX3 mRNA levels, however no significant differences between SOX3 and SOX9 mRNA levels were observed in the gonads of males that were kept under non-reproductive conditions compared to males kept with females under reproductive conditions and that were nest-builders.展开更多
Folliculogenesis is essential for production of female gametes in vertebrates.However,the molecular mechanisms underlying follicle development,particularly apoptosis regulation in ovary,remain elusive.Here,we gen erat...Folliculogenesis is essential for production of female gametes in vertebrates.However,the molecular mechanisms underlying follicle development,particularly apoptosis regulation in ovary,remain elusive.Here,we gen erated sox3 knockout zebrafish lines using CRISPR/Cas9.sox3 knockout led to follicle development retardation and a reduced fecundity in females.Comparative analysis of transcriptome between sox3^-/-and wild-type ovaries revealed that Sox3 was invoIved in pathways of ovarian steroidogenesis and apoptosis.Knockout of sox3 promoted follicle apoptosis and obvious apoptosis signals were detected in somatic cells of stages III and IV follicles of sox3^-/-ovaries.Moreover,Sox3 can bind to and activate the promoter of cyp19a1a.Up-regulation of Cyp19a1a expression promoted 17β-estradiol synthesis,which inhibited apoptosis in follicle development.Thus,Sox3 functions as a regulator of Cyp19a1a expression,via 17β-E2 linking apoptosis suppression,which is implicated in improving female fecundity.展开更多
Numerous studies have showed that sox3 is involved in neurogenesis and sex differentiation in vertebrates.However,the accurate expression pattern is still unclear in fish.In this study,medaka sox3 was isolated and its...Numerous studies have showed that sox3 is involved in neurogenesis and sex differentiation in vertebrates.However,the accurate expression pattern is still unclear in fish.In this study,medaka sox3 was isolated and its expression patterns were compared with germ cell gene vasa in adult gonads.By reverse transcription-polymerase chain reaction(RT-PCR)analysis,sox3 RNA expression was detected in the brain,eyes and gonads.By chromogenic and fluorescent in situ hybridization,sox3 was present in the oogonia and early stages of oocytes as well as granulosa cells and theca cells in the later stages;in the testis,sox3 was occurred in the Sertoli cells and sperm with a specific signal in the location of chromatid body of sperm;in the embryos,sox3 was expressed in the central nervous.These results suggest that medaka sox3 gene was occurred in somatic and germ cells in adult gonads of both sexes,involved in the process of spermatogenesis,as well as the development of the nervous system.This study provides a precise expression pattern of sox3 and suggests that sox3 may be involved in gonadal and nervous system development of fish.展开更多
文摘为初步阐明拟赤梢鱼(Pseudaspius leptocephalus)卵巢发育特征及sox3(sry related high mobility group box 3)基因在其卵巢发育过程中的作用,本研究通过组织切片观察了拟赤梢鱼卵巢早期发育过程,利用RACE技术克隆了该鱼sox3基因c DNA全长序列,并进行生物信息学分析;利用实时荧光定量PCR技术分析了sox3基因在拟赤梢鱼不同组织及性腺不同发育时期的表达模式。结果显示,拟赤梢鱼卵巢在孵化后45 d分化形成,160 d时由Ⅰ期进入Ⅱ期,孵化后360 d,卵巢仍处于Ⅱ期。拟赤梢鱼sox3基因cDNA全长为1800 bp(GenBank登录号:MT952206),编码299个氨基酸,存在保守的HMG(histidine,methionine,glycine-rich)结构域。氨基酸序列比对和系统进化树分析结果显示,拟赤梢鱼SOX3蛋白与斑马鱼(Danio rerio)和翘嘴鲌(Culter alburnus)亲缘关系最近。此外,sox3基因在拟赤梢鱼卵巢中表达量最高,其次是脑和眼睛,在精巢和其他组织中微量表达;在性腺分化过程中,sox3基因在卵巢中表达量显著高于未分化性腺和精巢,在卵巢发育阶段表达量不断升高,而在精巢中持续低水平表达。综上,本研究推测sox3基因主要参与拟赤梢鱼卵巢的分化和发育。
基金supported by the National Natural Science Foundation of China (No. 31372511)
文摘Evolutionary evidence suggests that Sox3, a member of the high-mobility-group(HMG) family of transcription factors, is an ancestral precursor of Sry and is involved in sex determination similar to Sry. However, there is limited information regarding the SOX3 gene of the black rockfish(Sebastes schlegeli). In this study, we first isolated SOX3 gene from the gonads of S. schlegeli by homology cloning. The full-length of S. schlegeli SOX3(SsSOX3) c DNA was 1386 bp, comprising a 906-bp open reading frame, which encodes a peptide showing 93.6% and 93.9% homology with the Sox3 proteins of Epinephelus coioides and Oryzias latipe, respectively. Comparison of the cDNA sequence of the Ss SOX3 gene with the corresponding genomic DNA fragment revealed that the SsSOX3 gene consists of a single exon. Phylogenetic analysis demonstrated the evolutionary relationship of Ss SOX3 with other known SOXB1 genes in fish and tetrapods. The promoter region contains binding sites of several transcriptional factors that might participate in the regulation of Ss SOX3 expression. Quantitative real-time PCR analysis indicated that SsSOX3 was expressed in all the investigated larval developmental stages from 1 to 35 days after birth and the level of expression gradually decreased as the development proceeded. SsSOX3 exhibited sexually dimorphic expression in adult gonads, with high expression in the ovary but low expression in the testis. In situ hybridization revealed that SsSOX3 was strongly expressed in oocytes and follicular cells of ovaries but slightly expressed in germ cells of testicular tissues. Therefore, this study suggests that Ss SOX3 plays an important role in oogenesis and ovary differentiation in S. schlegeli.
文摘In vertebrates, SOX (SRY-related HMG box) genes are thought to be due to major gene duplication events, initially occurring during early stages of metazoan evolution and later during the transition between non-vertebrate chordates and vertebrates. The aim of this study is to examine SOX3 and SOX9 transcription in oogensis and spermatogenesis in a fish model, the blue gourami (Trichogaster trichopterus). In females during oogenesis, SOX9 mRNA levels were lower compared to SOX3 mRNA levels. In males, SOX9 mRNA levels were higher in testes compared to SOX3 mRNA levels, however no significant differences between SOX3 and SOX9 mRNA levels were observed in the gonads of males that were kept under non-reproductive conditions compared to males kept with females under reproductive conditions and that were nest-builders.
基金the National Natural Science Foundation of China and National Key Technologies R&D Program.
文摘Folliculogenesis is essential for production of female gametes in vertebrates.However,the molecular mechanisms underlying follicle development,particularly apoptosis regulation in ovary,remain elusive.Here,we gen erated sox3 knockout zebrafish lines using CRISPR/Cas9.sox3 knockout led to follicle development retardation and a reduced fecundity in females.Comparative analysis of transcriptome between sox3^-/-and wild-type ovaries revealed that Sox3 was invoIved in pathways of ovarian steroidogenesis and apoptosis.Knockout of sox3 promoted follicle apoptosis and obvious apoptosis signals were detected in somatic cells of stages III and IV follicles of sox3^-/-ovaries.Moreover,Sox3 can bind to and activate the promoter of cyp19a1a.Up-regulation of Cyp19a1a expression promoted 17β-estradiol synthesis,which inhibited apoptosis in follicle development.Thus,Sox3 functions as a regulator of Cyp19a1a expression,via 17β-E2 linking apoptosis suppression,which is implicated in improving female fecundity.
基金the National Key R&D Program of China(2018YFD0901205)National Natural Science Foundation of China(31672700).
文摘Numerous studies have showed that sox3 is involved in neurogenesis and sex differentiation in vertebrates.However,the accurate expression pattern is still unclear in fish.In this study,medaka sox3 was isolated and its expression patterns were compared with germ cell gene vasa in adult gonads.By reverse transcription-polymerase chain reaction(RT-PCR)analysis,sox3 RNA expression was detected in the brain,eyes and gonads.By chromogenic and fluorescent in situ hybridization,sox3 was present in the oogonia and early stages of oocytes as well as granulosa cells and theca cells in the later stages;in the testis,sox3 was occurred in the Sertoli cells and sperm with a specific signal in the location of chromatid body of sperm;in the embryos,sox3 was expressed in the central nervous.These results suggest that medaka sox3 gene was occurred in somatic and germ cells in adult gonads of both sexes,involved in the process of spermatogenesis,as well as the development of the nervous system.This study provides a precise expression pattern of sox3 and suggests that sox3 may be involved in gonadal and nervous system development of fish.