Sox9在脊椎动物性腺分化及精巢发育中行使重要功能。为探究线纹海马sox9a的分子特征及表达模式,本研究采用RACE获得了其cDNA全长1979 bp,包含339 bp 5’ UTR、1488 bp的ORF和152 bp 3’ UTR,共编码496个氨基酸;氨基酸多重比对显示,线纹...Sox9在脊椎动物性腺分化及精巢发育中行使重要功能。为探究线纹海马sox9a的分子特征及表达模式,本研究采用RACE获得了其cDNA全长1979 bp,包含339 bp 5’ UTR、1488 bp的ORF和152 bp 3’ UTR,共编码496个氨基酸;氨基酸多重比对显示,线纹海马sox9具有高度保守的HMG-box结构域;系统进化树分析表明,其与侏儒海马sox9a的亲缘关系最近;RT-qPCR显示,sox9a泛在表达于线纹海马成体的主要组织中,在性腺中呈显著的雄性偏好性高表达于精巢,揭示其可能参与线纹海马精巢的分化及维持。本研究为后续深入研究线纹海马sox9a在性别发育中的功能奠定了基础。展开更多
DEAR EDITOR,Sox9 is a member of the Sry-related high-mobility group box(Sox)transcription factor family in animals.In teleost fish,Sox9 undergoes duplication to generate two duplicates,namely Sox9a and Sox9b.However,t...DEAR EDITOR,Sox9 is a member of the Sry-related high-mobility group box(Sox)transcription factor family in animals.In teleost fish,Sox9 undergoes duplication to generate two duplicates,namely Sox9a and Sox9b.However,the functions of these duplicates in the teleost Nile tilapia(Oreochromis niloticus)remain unclear.In this study,we characterized the roles of Nile tilapia Sox9a in chondrogenesis and gonadal development.In situ hybridization assays showed that Sox9a was mainly expressed in cartilage tissues and somatic cells surrounding germ cells of the gonads.CRISPR/Cas9-mediated homozygous mutation of the Sox9a gene resulted in craniofacial deformities and missed mandibles,as well as impaired the expression of Col2a1a that is involved in chondrogenesis.In addition,germ cell number and DNA replication in somatic cells in the gonads of both sexes were reduced following Sox9a mutation.Taken together,this study demonstrates that Sox9a is involved in cartilage development and germ cell proliferation in Nile tilapia.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
Two transcription factors, Sox9a and Foxl2 were cloned from half-smooth tongue sole （Cynoglossus semilaevis）. Sox9a is a new duplication of C.semilaevis Sox9 gene. The complete cDNA of Sox9a gene was 1 842 bp long c...Two transcription factors, Sox9a and Foxl2 were cloned from half-smooth tongue sole （Cynoglossus semilaevis）. Sox9a is a new duplication of C.semilaevis Sox9 gene. The complete cDNA of Sox9a gene was 1 842 bp long coding for 487 amino acids and Foxl2 gene was 1 817 bp coding for 308 amino acids. Sox9a was expressed higher in male brain, pituitary and gonad and Foxl2 were higher in female brain, pituitary and gonad. The expression of Sox9a gene in gonads of neo-males was higher than that of normal females. Sox9a and Foxl2 were expressed higher in gastrula stage than in other stages. In the period of sex differentiation, the expression of Sox9a was first going up and then going down and Foxl2 was higher expressed at 37 dph. The highest expressions of Sox9a and Foxl2 genes occurred in nine-month and 12-month old gonad tissues, respectively. Sox9a gene was considered to have inevitable links with sex reversal, sex differentiation and cell differentiation of embryos and formation of spermatogenic ceils. Foxl2 was considered to play a role in sex differentiation, cell differentiation of embryos but not to be necessary for sex determination and sex reversal.展开更多
Both glial cells and glia scar greatly affect the development of spinal cord injury and have become hot spots in research on spinal cord injury treatment.The cellular deposition of dense extracellular matrix proteins ...Both glial cells and glia scar greatly affect the development of spinal cord injury and have become hot spots in research on spinal cord injury treatment.The cellular deposition of dense extracellular matrix proteins such as chondroitin sulfate proteoglycans inside and around the glial scar is known to affect axonal growth and be a major obstacle to autogenous repair.These proteins are thus candidate targets for spinal cord injury therapy.Our previous studies demonstrated that 810 nm photo biomodulation inhibited the formation of chondroitin sulfate proteoglycans after spinal cord injury and greatly improved motor function in model animals.However,the specific mechanism and potential targets involved remain to be clarified.In this study,to investigate the therapeutic effect of photo biomodulation,we established a mouse model of spinal cord injury by T9 clamping and irradiated the injury site at a power density of 50 mW/cm~2 for 50 minutes once a day for 7 consecutive days.We found that photobiomodulation greatly restored motor function in mice and down regulated chondroitin sulfate proteoglycan expression in the injured spinal cord.Bioinformatics analysis revealed that photobiomodulation inhibited the expression of proteoglycan-related genes induced by spinal cord injury,and versican,a type of proteoglycan,was one of the most markedly changed molecules.Immunofluorescence staining showed that after spinal cord injury,versican was present in astrocytes in spinal cord tissue.The expression of versican in primary astrocytes cultured in vitro increased after inflammation induction,whereas photobiomodulation inhibited the expression of ve rsican.Furthermore,we found that the increased levels of p-Smad3,p-P38 and p-Erk in inflammatory astrocytes were reduced after photobiomodulation treatment and after delivery of inhibitors including FR 180204,(E)-SIS3,and SB 202190.This suggests that Sma d 3/Sox9 and MAP K/Sox9 pathways may be involved in the effects of photobiomodulation.In summary,our findings show that photobiomodulation modulates the expression of chondroitin sulfate proteoglycans,and versican is one of the key target molecules of photo biomodulation.MAPK/Sox9 and Smad3/Sox9 pathways may play a role in the effects of photo biomodulation on chondroitin sulfate proteoglycan accumulation after spinal cord injury.展开更多
Anti-Mullerian hormone （amh） is one of the earliest functional genes expressed during testicular differentiation. It has been suggested that androgen signaling regulates critical genes for the differentiation and de...Anti-Mullerian hormone （amh） is one of the earliest functional genes expressed during testicular differentiation. It has been suggested that androgen signaling regulates critical genes for the differentiation and development of the testis. To elucidate the exact regulatory mechanisms involved in arnh transcription mediated by androgen signaling, androgen signaling was manipulated in zebrafish by cytochrome P450 17al （cyplTal） knockout and Flutamide treatment. In cyp17a1-deficient and Flutamide-treated testes, up-regulated sry-box9a （soxga） and down-regulated amh were observed. Moreover, a physical association of the zebrafish androgen receptor （AR） and SOX9A was found. The interaction between AR and SOX9A was mediated via the DNA binding domain （DBD） of AR and the transactivation domain （TA） of SOX9A, and was further enhanced by 5-alpha dihydrotestosterone （DHT）, one of the most potent androgens. Intriguingly, together with SOX9A, androgen signaling synergistically promoted amh transcription, mainly through the proximal 1 kb of the amh promoter region. Taken together, our data demonstrate a critical mechanism underlying the direct synergy of androgen signaling and SOX9A in the regulation of amh transcription.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
文摘Sox9在脊椎动物性腺分化及精巢发育中行使重要功能。为探究线纹海马sox9a的分子特征及表达模式,本研究采用RACE获得了其cDNA全长1979 bp,包含339 bp 5’ UTR、1488 bp的ORF和152 bp 3’ UTR,共编码496个氨基酸;氨基酸多重比对显示,线纹海马sox9具有高度保守的HMG-box结构域;系统进化树分析表明,其与侏儒海马sox9a的亲缘关系最近;RT-qPCR显示,sox9a泛在表达于线纹海马成体的主要组织中,在性腺中呈显著的雄性偏好性高表达于精巢,揭示其可能参与线纹海马精巢的分化及维持。本研究为后续深入研究线纹海马sox9a在性别发育中的功能奠定了基础。
基金supported by the National Key Research and Development Program of China(2022YFD1201600)National Natural Science Foundation of China(31861123001,31302170,and 31772831)+1 种基金Chongqing Science and Technology Bureau(CSTB2022NSCQ-MSX1608 and cstc2021ycjh-bgzxm0024)Chongqing Fishery Technology Innovation Union(2023)。
文摘DEAR EDITOR,Sox9 is a member of the Sry-related high-mobility group box(Sox)transcription factor family in animals.In teleost fish,Sox9 undergoes duplication to generate two duplicates,namely Sox9a and Sox9b.However,the functions of these duplicates in the teleost Nile tilapia(Oreochromis niloticus)remain unclear.In this study,we characterized the roles of Nile tilapia Sox9a in chondrogenesis and gonadal development.In situ hybridization assays showed that Sox9a was mainly expressed in cartilage tissues and somatic cells surrounding germ cells of the gonads.CRISPR/Cas9-mediated homozygous mutation of the Sox9a gene resulted in craniofacial deformities and missed mandibles,as well as impaired the expression of Col2a1a that is involved in chondrogenesis.In addition,germ cell number and DNA replication in somatic cells in the gonads of both sexes were reduced following Sox9a mutation.Taken together,this study demonstrates that Sox9a is involved in cartilage development and germ cell proliferation in Nile tilapia.
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The National High Technology Research and Development Program of China (863 Program) under contract No.2006AA10A401the Taishan Scholar Project of Shandong Province,Chinathe National Natural Science Foundation of China under contract Nos 41006107 and 30972244
文摘Two transcription factors, Sox9a and Foxl2 were cloned from half-smooth tongue sole （Cynoglossus semilaevis）. Sox9a is a new duplication of C.semilaevis Sox9 gene. The complete cDNA of Sox9a gene was 1 842 bp long coding for 487 amino acids and Foxl2 gene was 1 817 bp coding for 308 amino acids. Sox9a was expressed higher in male brain, pituitary and gonad and Foxl2 were higher in female brain, pituitary and gonad. The expression of Sox9a gene in gonads of neo-males was higher than that of normal females. Sox9a and Foxl2 were expressed higher in gastrula stage than in other stages. In the period of sex differentiation, the expression of Sox9a was first going up and then going down and Foxl2 was higher expressed at 37 dph. The highest expressions of Sox9a and Foxl2 genes occurred in nine-month and 12-month old gonad tissues, respectively. Sox9a gene was considered to have inevitable links with sex reversal, sex differentiation and cell differentiation of embryos and formation of spermatogenic ceils. Foxl2 was considered to play a role in sex differentiation, cell differentiation of embryos but not to be necessary for sex determination and sex reversal.
基金supported by the National Natural Science Foundation of China,Nos.81070996(to ZW),81572151(to XH)Shaanxi Provincial Key R&D Program,Nos.2020ZDLSF02-05(to ZW),2021ZDLSF02-10(to XH)+1 种基金Everest Project of Military Medicine of Air Force Medical University,No.2018RCFC02(to XH)Boosting Project of the First Affiliated Hospital of Air Force Medical University,No.XJZT19Z22(to ZW)。
文摘Both glial cells and glia scar greatly affect the development of spinal cord injury and have become hot spots in research on spinal cord injury treatment.The cellular deposition of dense extracellular matrix proteins such as chondroitin sulfate proteoglycans inside and around the glial scar is known to affect axonal growth and be a major obstacle to autogenous repair.These proteins are thus candidate targets for spinal cord injury therapy.Our previous studies demonstrated that 810 nm photo biomodulation inhibited the formation of chondroitin sulfate proteoglycans after spinal cord injury and greatly improved motor function in model animals.However,the specific mechanism and potential targets involved remain to be clarified.In this study,to investigate the therapeutic effect of photo biomodulation,we established a mouse model of spinal cord injury by T9 clamping and irradiated the injury site at a power density of 50 mW/cm~2 for 50 minutes once a day for 7 consecutive days.We found that photobiomodulation greatly restored motor function in mice and down regulated chondroitin sulfate proteoglycan expression in the injured spinal cord.Bioinformatics analysis revealed that photobiomodulation inhibited the expression of proteoglycan-related genes induced by spinal cord injury,and versican,a type of proteoglycan,was one of the most markedly changed molecules.Immunofluorescence staining showed that after spinal cord injury,versican was present in astrocytes in spinal cord tissue.The expression of versican in primary astrocytes cultured in vitro increased after inflammation induction,whereas photobiomodulation inhibited the expression of ve rsican.Furthermore,we found that the increased levels of p-Smad3,p-P38 and p-Erk in inflammatory astrocytes were reduced after photobiomodulation treatment and after delivery of inhibitors including FR 180204,(E)-SIS3,and SB 202190.This suggests that Sma d 3/Sox9 and MAP K/Sox9 pathways may be involved in the effects of photobiomodulation.In summary,our findings show that photobiomodulation modulates the expression of chondroitin sulfate proteoglycans,and versican is one of the key target molecules of photo biomodulation.MAPK/Sox9 and Smad3/Sox9 pathways may play a role in the effects of photo biomodulation on chondroitin sulfate proteoglycan accumulation after spinal cord injury.
基金supported by the National Natural Science Foundation of China (31501857 to G.Z.and 31530077 to Z.Y.)the National Basic Research Program of China (2014CB138602 to Z.Y.)
文摘Anti-Mullerian hormone （amh） is one of the earliest functional genes expressed during testicular differentiation. It has been suggested that androgen signaling regulates critical genes for the differentiation and development of the testis. To elucidate the exact regulatory mechanisms involved in arnh transcription mediated by androgen signaling, androgen signaling was manipulated in zebrafish by cytochrome P450 17al （cyplTal） knockout and Flutamide treatment. In cyp17a1-deficient and Flutamide-treated testes, up-regulated sry-box9a （soxga） and down-regulated amh were observed. Moreover, a physical association of the zebrafish androgen receptor （AR） and SOX9A was found. The interaction between AR and SOX9A was mediated via the DNA binding domain （DBD） of AR and the transactivation domain （TA） of SOX9A, and was further enhanced by 5-alpha dihydrotestosterone （DHT）, one of the most potent androgens. Intriguingly, together with SOX9A, androgen signaling synergistically promoted amh transcription, mainly through the proximal 1 kb of the amh promoter region. Taken together, our data demonstrate a critical mechanism underlying the direct synergy of androgen signaling and SOX9A in the regulation of amh transcription.
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.
