Oryzias celebensis is determined by an XX-XY system,however,its sex-related genes have not been confirmed.The genes foxl2 and sox9b are important for gonadal development in vertebrates.In the present study,the foxl2 a...Oryzias celebensis is determined by an XX-XY system,however,its sex-related genes have not been confirmed.The genes foxl2 and sox9b are important for gonadal development in vertebrates.In the present study,the foxl2 and sox9b cDNAs were cloned and their expression patterns were analyzed.The cDNA sequence of foxl2 and sox9b were obtained by rapid amplification of cDNA ends(RACEs).Reverse transcription polymerase chain reaction(RT-PCR)showed that O.celebensis foxl2(Ocfoxl2)transcript was predominately expressed in the ovary while O.celebensis sox9b(Ocsox9b)transcript was mainly expressed in the testis.In embryos,Ocfoxl2 transcript was first detected in gastrula.In contrast,Ocsox9b transcript was of maternal origin as expression was detected at the two cell stage.By chromogenic and fluorescent in situ hybridization(ISH),Ocfoxl2 mRNA in the ovary was highly expressed in early oocytes and weakly expressed in the granulosa cells and thecal cells in the later oocytes.In contrast,Ocsox9b mRNA was mainly found in Sertoli cells surrounding spermatogonia and in spermatids and sperm in the testis.In summary,Ocfoxl2 identified somatic cells and differentiating oocytes in the ovary,indicating it is involved in ovarian development.Ocsox9b is identified in Sertoli cells and late developing male germ cells indicating it is involved in testicular development.展开更多
Sox9 is a multifunctional gene and plays crucial roles in vertebrate development including chondrogensis.In teleost,due to the genome duplication event,there are two co-orthologs sox9a and sox9b.In this study,CRISPR/C...Sox9 is a multifunctional gene and plays crucial roles in vertebrate development including chondrogensis.In teleost,due to the genome duplication event,there are two co-orthologs sox9a and sox9b.In this study,CRISPR/Cas9 technology was performed to disrupt the function of either sox9a or sox9b.All sox9a mutants(sox9aΔ10 and sox9aΔ67)and sox9b mutants(sox9bΔ11 and sox9bΔ20)lost HMG domain and Q/S domain,however,only sox9a mutant larvae had mis-shaped pectoral fins and lacked the scapulocoracoid cartilage.sox9b mutant larvae showed normal cartilages similar to wild type larvae.The results suggested that sox9a,not sox9b was required for cartilage development in zebrafish,which was different from the sox9b-mutant phenotype induced by N-ethyl-N-nitrosourea(ENU)treatment,gamma radiation treatment or morpholino injection.This study confirmed that ancestral sox9 gene functions partitioned between the two paralogs in zebrafish.展开更多
为探究sox9基因在大黄鱼性别决定与分化过程中的作用,利用RACE方法克隆得到大黄鱼sox9a和sox9b 2个基因,采用实时荧光定量PCR(qRT-PCR)分析其在雌雄个体不同组织和不同发育阶段的表达规律,并检测了其在17β-雌二醇处理后的遗传雄性和17...为探究sox9基因在大黄鱼性别决定与分化过程中的作用,利用RACE方法克隆得到大黄鱼sox9a和sox9b 2个基因,采用实时荧光定量PCR(qRT-PCR)分析其在雌雄个体不同组织和不同发育阶段的表达规律,并检测了其在17β-雌二醇处理后的遗传雄性和17α-甲基睾酮诱导处理后遗传雌性幼鱼中的表达变化。结果显示,大黄鱼sox9a c DNA全长2 442 bp(NCBI登录号:MH996431),包含476 bp 5′UTR、466 bp 3′UTR、1 500 bp ORF,编码499个氨基酸。大黄鱼sox9b cDNA全长为2 199 bp(NCBI登录号:MH996432),包含335 bp5′UTR、415 bp 3′UTR、1 449 bp ORF,编码482个氨基酸。qRT-PCR分析显示,sox9a基因主要在性腺、眼、脑、肝脏中表达,精巢中的表达量显著高于卵巢;sox9b在大黄鱼各组织中广泛表达,但以精巢中表达量最高,而卵巢中只有痕量表达。在鱼苗性腺发育初期,sox9a/b的表达量都维持在较低水平;随着鱼苗长大,sox9a/b的表达量在84 dph、123 dph 2次达到高峰,随后开始下降,10 mph后又逐渐上升。17β-雌二醇处理能够显著下调遗传雄性大黄鱼sox9a和sox9b基因在性腺中的表达,17α-甲基睾酮处理则显著上调遗传雌性大黄鱼sox9a和sox9b基因在性腺中的表达。研究表明,sox9a/b基因与大黄鱼性别发育和分化过程密切相关,对大黄鱼精巢的发育与维持起重要的调控作用,而且两个基因的功能可能具有一定程度的分化。展开更多
Retinoic acid (RA) plays an important role in development of vertebrate embryos. We demonstrate impacts of exogenous RA on the formation of neural crest cells in zebrafish using specific neural crest markers sox9b and...Retinoic acid (RA) plays an important role in development of vertebrate embryos. We demonstrate impacts of exogenous RA on the formation of neural crest cells in zebrafish using specific neural crest markers sox9b and crestin. Treatment with all-trans RA at 10-7 mmol/L at 50% epiboly induces sox9b expression in the forebrain and crestin expression in the forebrain and midbrain, resulting in significant increase of pigment cells in the head derived from the cranial neural crest. In addition, RA treatment induces expression of sox9b and crestin in the caudal marginal cells of the neuroectoderm during early segmentation. Earlier commitment of these cells to the neural crest fate in the posterior margins leads to abnormal development of the posterior body, probably by preventing mingling of ventral derived and dorsal-derived cells during the formation of the tailbud.展开更多
基金This work was supported by National Key R&D Program of China(2018YFD0901205)National Natural Science Foundation of China(31672700,31372520).
文摘Oryzias celebensis is determined by an XX-XY system,however,its sex-related genes have not been confirmed.The genes foxl2 and sox9b are important for gonadal development in vertebrates.In the present study,the foxl2 and sox9b cDNAs were cloned and their expression patterns were analyzed.The cDNA sequence of foxl2 and sox9b were obtained by rapid amplification of cDNA ends(RACEs).Reverse transcription polymerase chain reaction(RT-PCR)showed that O.celebensis foxl2(Ocfoxl2)transcript was predominately expressed in the ovary while O.celebensis sox9b(Ocsox9b)transcript was mainly expressed in the testis.In embryos,Ocfoxl2 transcript was first detected in gastrula.In contrast,Ocsox9b transcript was of maternal origin as expression was detected at the two cell stage.By chromogenic and fluorescent in situ hybridization(ISH),Ocfoxl2 mRNA in the ovary was highly expressed in early oocytes and weakly expressed in the granulosa cells and thecal cells in the later oocytes.In contrast,Ocsox9b mRNA was mainly found in Sertoli cells surrounding spermatogonia and in spermatids and sperm in the testis.In summary,Ocfoxl2 identified somatic cells and differentiating oocytes in the ovary,indicating it is involved in ovarian development.Ocsox9b is identified in Sertoli cells and late developing male germ cells indicating it is involved in testicular development.
基金This work was supported by the Fundamental Research Funds for the Central Universities(2662017PY013)
文摘Sox9 is a multifunctional gene and plays crucial roles in vertebrate development including chondrogensis.In teleost,due to the genome duplication event,there are two co-orthologs sox9a and sox9b.In this study,CRISPR/Cas9 technology was performed to disrupt the function of either sox9a or sox9b.All sox9a mutants(sox9aΔ10 and sox9aΔ67)and sox9b mutants(sox9bΔ11 and sox9bΔ20)lost HMG domain and Q/S domain,however,only sox9a mutant larvae had mis-shaped pectoral fins and lacked the scapulocoracoid cartilage.sox9b mutant larvae showed normal cartilages similar to wild type larvae.The results suggested that sox9a,not sox9b was required for cartilage development in zebrafish,which was different from the sox9b-mutant phenotype induced by N-ethyl-N-nitrosourea(ENU)treatment,gamma radiation treatment or morpholino injection.This study confirmed that ancestral sox9 gene functions partitioned between the two paralogs in zebrafish.
文摘为探究sox9基因在大黄鱼性别决定与分化过程中的作用,利用RACE方法克隆得到大黄鱼sox9a和sox9b 2个基因,采用实时荧光定量PCR(qRT-PCR)分析其在雌雄个体不同组织和不同发育阶段的表达规律,并检测了其在17β-雌二醇处理后的遗传雄性和17α-甲基睾酮诱导处理后遗传雌性幼鱼中的表达变化。结果显示,大黄鱼sox9a c DNA全长2 442 bp(NCBI登录号:MH996431),包含476 bp 5′UTR、466 bp 3′UTR、1 500 bp ORF,编码499个氨基酸。大黄鱼sox9b cDNA全长为2 199 bp(NCBI登录号:MH996432),包含335 bp5′UTR、415 bp 3′UTR、1 449 bp ORF,编码482个氨基酸。qRT-PCR分析显示,sox9a基因主要在性腺、眼、脑、肝脏中表达,精巢中的表达量显著高于卵巢;sox9b在大黄鱼各组织中广泛表达,但以精巢中表达量最高,而卵巢中只有痕量表达。在鱼苗性腺发育初期,sox9a/b的表达量都维持在较低水平;随着鱼苗长大,sox9a/b的表达量在84 dph、123 dph 2次达到高峰,随后开始下降,10 mph后又逐渐上升。17β-雌二醇处理能够显著下调遗传雄性大黄鱼sox9a和sox9b基因在性腺中的表达,17α-甲基睾酮处理则显著上调遗传雌性大黄鱼sox9a和sox9b基因在性腺中的表达。研究表明,sox9a/b基因与大黄鱼性别发育和分化过程密切相关,对大黄鱼精巢的发育与维持起重要的调控作用,而且两个基因的功能可能具有一定程度的分化。
基金This work was supported by the National Natural Science Foundation of China (Grant Nos. 30025020 and 39970360)TRAPOYT of the MOE.
文摘Retinoic acid (RA) plays an important role in development of vertebrate embryos. We demonstrate impacts of exogenous RA on the formation of neural crest cells in zebrafish using specific neural crest markers sox9b and crestin. Treatment with all-trans RA at 10-7 mmol/L at 50% epiboly induces sox9b expression in the forebrain and crestin expression in the forebrain and midbrain, resulting in significant increase of pigment cells in the head derived from the cranial neural crest. In addition, RA treatment induces expression of sox9b and crestin in the caudal marginal cells of the neuroectoderm during early segmentation. Earlier commitment of these cells to the neural crest fate in the posterior margins leads to abnormal development of the posterior body, probably by preventing mingling of ventral derived and dorsal-derived cells during the formation of the tailbud.
