用户名: 密码: 验证码:
罗非鱼和南方鲇生殖细胞几种分子标记的克隆和表达研究
详细信息    本馆镜像全文|  推荐本文 |  |   获取CNKI官网全文
摘要
性别决定和分化一直是生物学、生理学、内分泌学研究的热点问题之一。鱼类的性别在很大程度上受到环境因素如:温度、光照、外源性激素水平的影响,甚至还能导致性逆转。过去,对于性别分化的研究主要集中在对性腺体细胞的分化方面,相继克隆了许多在鱼类性别决定和分化中起重要作用的基因,并对其功能进行了深入研究。近年来,由于对性别分化的认识不断加深,加上鱼类养殖的要求,对生殖细胞的分化和发育逐渐成为一个研究的热点。对鱼类生殖细胞标记(germ cell marker)基因进行研究将为深入开展对鱼类生殖细胞分化和发育分子机制研究,拓展对这一领域的认识奠定坚实基础,也对水产养殖业具有指导意义。本研究所用模式动物尼罗罗非鱼(Oreochromis niloticus)具有生长速度快、性成熟时间快、繁殖周期短等特点,是世界最重要的养殖鱼类之一。利用现代生物学手段对其进行单性化的养殖提出了新的要求。因此对罗非鱼性别分化,特别是生殖细胞的分化和发育进行研究具有重要的理论意义和应用价值。
     本实验从尼罗罗非鱼克隆了几种生殖细胞标记,研究了它们的组织表达模式和在个体发生中的表达。同时我们还利用fadrozole(芳香化酶抑制剂)、雌二醇处理,人工诱导雌雄性逆转罗非鱼,研究了生殖细胞标记在这些性逆转鱼性腺中的表达变化情况。
     采用RT-PCR和RACE相结合的方法,克隆了两个尼罗罗非鱼早期发育阶段的生殖细胞标记:Figla和Oct4。克隆得到的Figla序列长度为1254 bp,开放阅读框(open reading frame,ORF)长度为594碱基(base pair,bp),编码197个氨基酸(amino acid,aa)。Oct4序列长度为1787 bp,ORF长度为1380 bp,编码459个aa。它们在卵巢中的表达均高于精巢。罗非鱼Figla(ntFigla)m RNA从孵化后第5天(days after hatching,dah)开始表达于雌性和雄性性腺中,并从10 dah开始呈现性别二态性表达,RT-PCR和定量RT-PCR(quantitative RT-PCR,qRT-PCR)都显示,直到70 dah ntFigla在精巢的mRNA水平都维持较低水平。而性成熟后,它在精巢的表达明显增加。ntOct4 mRNA在性腺发育的早期高表达,这暗示它在这个阶段可能具有重要功能。但在孵化后1个月通过原位杂交可以检测到它的性别二态性表达,之后它在卵巢中持续高水平表达,而在精巢中表达量逐渐降低,直到检测不到。
     采用RT-PCR和RACE相结合的方法,我们从尼罗罗非鱼克隆了两个与生殖细胞减数分裂相关的基因:Spo11和Scp3。Spo11是最早分离得到的减数分裂重组基因之一,而Scp3是减数分裂联会复合体(Synaptonemal complex,SC)的主要结构蛋白,它们均为减数分裂特异的标记。Scp3的cDNA序列为1070 bp,ORF为717 bp,编码238个aa;尼罗罗非鱼Spo11序列为1468 bp,ORF为1173 bp,编码390个aa。组织分布结果表明,两者都在罗非鱼性腺中特异表达,ntScp3主要表达于精巢,而ntSpo11主要表达于卵巢。它们都能用作检测处于减数分裂的生殖细胞。在个体发生研究结果表明,Scp3 mRNA的表达总是精巢高于卵巢,这种差异一直持续到性成熟。在卵巢中,ntScp3 mRNA主要在Ⅰ、Ⅱ、Ⅲ时相的卵母细胞中表达,在第Ⅳ时相的卵母细胞中表达量低,且主要集中在细胞边缘。ntSpo11主要在卵巢中表达,这种性别差异均能在不同时期(10 dah、30 dah、40 dah、5个月和成体)检测到。
     Vasa蛋白是DEAD-box蛋白家族成员中一种ATP依赖的RNA解旋酶,它在高等后生动物的生殖细胞形成中起着关键的作用。Vasa基因特异性地表达于许多后生动物的生殖细胞中,并被广泛确定为许多脊椎动物和无脊椎动物生殖细胞的特异标记。采用RT-PCR和RACE相结合的方法,从南方鲇中分离得到Vasa基因的两个亚型,scVasa和scVasa-s。通过RACE方法获得了它们的全长。核苷酸序列分析显示,scVasa cDNA序列全长2,563bp,其中开放阅读框ORF长1,989bp,可以编码含662个氨基酸的蛋白;scVasa-s cDNA序列全长2,475bp,其中开放阅读框ORF长1,926bp,可以编码含641个氨基酸的蛋白。scVasa-s缺乏scVasa N末端区域的一段序列。这两种推导出的氨基酸序列均具有DEAD-box蛋白家族成员的特征,即含有4个精氨酸-甘氨酸-甘氨酸(RGG)基序和另外8个保守基序。它们和银鲫的Vasa同源蛋白有很高的相似性(75.2%和73.8%)。原位杂交结果表明:在卵巢,scVasa主要在Ⅰ、Ⅱ时相的卵母细胞表达,而在精巢,主要在精原细胞和初级精母细胞表达。半定量PCR结果显示,在生殖周期中,两种亚型在以Ⅱ时相卵母细胞为主体的卵巢恢复期表达均高于以Ⅲ-Ⅳ时相卵母细胞为主体的卵黄生成期。
     本研究首次从硬骨鱼类罗非鱼和南方鲇系统的克隆了生殖细胞和减数分裂的几种分子标记。尽管目前国内外众多学者对生殖细胞和减数分裂的分子标记进行了深入广泛的研究,但是大都集中在模式生物如小鼠、斑马鱼和青鳉上,而对具有重要经济意义的水产养殖鱼类的研究却很少,甚至是空白,因而本研究通过对我国重要的养殖鱼类罗非鱼和南方鲇的生殖细胞和减数分裂的分子标记的表达模式以及它们在药物处理性逆转过程中的表达变化,为最终通过生物技术手段解释性别分化的机制、对减数分裂起始的调控机制、体细胞和生殖细胞的相互关系的研究奠定基础,而且在未来的水产养殖业中有非常广阔的应用前景。
Sex determination and differentiation have always been the focus in the researches of biology, physiology and endocrinology.In mammals,sex was controlled by genetic factors,especially the sex determining gene,SRY/Sry.While in lower vertebrates,including fish,sex is also influenced by environmental factors,such as temperature,photoperiod and exogenous sex steroids.These factors might even cause sex reversal in many species.Sex differentiation includes the differentiation of both somatic cells and germ cells.In the past,much attention has been drawn to the differentiation of somatic cells,genes closely related to the sex determination and differentiation have been cloned and their functions further investigated.Recently,researches on the differentiation of germ cells have gradually become a new hotspot based on the better understanding of sex differentiation and requirement of fishery industry.To date,researches of germ cells have been mostly restricted in mammals,while in lower vertebrates,especially in fish,the germ cells were poorly investigated.One of the most important reasons responsible for this is due to the lack of germ cell markers. Investigations on the germ cell marker genes in fish will lay foundations for the researches on the mechanisms of germ cell differentiation and development,and expanding the understanding of sex differentiation in fish.It also has practical implications for aquaculture.
     The Nile tilapia(Oreochromis niloticus),because of its fast growth rate and short reproductive cycle,is developed as one of the most important species for aquaculture and experiment globally.As the growth of male is much faster than female by 50%,males are always favored.Because the masculinization methods generally utilized was of much disadvantages,new techniques by modern molecular biology is highly required in growing monosex progeny.As a result,investigations on the sex differentiation,especially on the differentiatioin and development of germ cells,are of decisive theoretical significance and practical implications.
     In the present study,several germ cell markers were cloned and their expressions in various adult tissues and ontogenic expressions in gonad were investigated.Sex reversals were successfully obatained by treatment with aromatase inhitor(fadrozole) and estrogen(17-βestrodial).The expressions of these marker genes in the gonads of sex reversed fish were also examined.
     First of all,two cDNAs of germ cell markers(Oct4 and Figla) responsible for early development of tilapia gonad were isolated by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends(RACE).The length of Figla cDNA is 1254bp, containing an open reading frame(ORF) of 594bp,which encode a 198aa protein.The length of Oct4 cDNA is 1787bp,containing an ORF of 1380bp,which encode a 460aa protein.
     The expression of Figla mRNA initiated from the 5 days after hatching(dab) and showed a sexual dimorphic pattern from 10 dah onwards.The expression level of Figla mRNA in testis remained low till 70 dah,while increased significant when sexually matured detected by both RT-PCR and quantitative RT-PCR(qRT-PCR).High expression of ntOct4 was detected during early gonadal development in both sexes,indicating its decisive role in this period.In situ hybridization(ISH) results revealed that ntOct4 showed a sexual dimorphic expression pattern, expressing higher in ovary than in testis from 1 month after hatching,after which,its expression level decreased significantly and became undetectable in the adult male.
     cDNAs of Spo11,one of the earliest marker related to recombination process in meiosis and Scp3 encoding the structural protein of synaptonemal complex during meiosis were also isolated by RT-PCR and RACE methods in the Nile tilapia.Both of them are markers closely related to the meiosis of germ cells.Full length cDNAs of Scp3 and Spo11 were 1070 and 1468 bp,which contain ORFs of 717 and 1173 bp,encoding proteins of 239 and 390 aa,respectively.Expressions of both markers were restricted to the gonads of the tilapia,with higher level in testis for Scp3 and higher level in ovary for Spo11.
     Tissue distribution analyse showed that the the expression of Scp3 was much higher in testis than in ovary.Ontogeny analysis by RT-PCR also revealed this pattern basically.It was also revealed that Scp3 was expressed mainly in theⅠ,ⅡandⅢphase oocytes and its expression level decreased in oocytes ofⅣphase,in which mainly concentrating on the edge.Spo11 expression also showed sexual dimorphic in tissue distribution analysis,being higher in ovary than in testis.Ontogeny analysis by qRT-PCR revealed the same pattern in 10 dah,30 dah,40 dah and 5 months fish and adults.
     Vasa protein,a member of the DEAD(Asp-Glu-Alu-Asp) box protein family of ATP-dependent RNA helicases,plays key roles in germ cell formation in higher metazoans.The Vasa gene,found exclusively expressed in germline cells of many metazoans,has been extensively characterized as germ cell specific marker in many vertebrate and invertebrate species.Two isoforms of Vasa cDNA (scVasa and it's short form scVasa-s) were isolated and characterized in a teleost fish,southern catfish by RT-PCR and RACE methods.Analysis of the nucleotide sequences revealed that the full length cDNA of scVasa comprises 2,522 bp with an ORF of 1,989 bp encoding 662 amino acids, while that of scVasa-s comprises 2,475 bp with an ORF of 1,926 bp encoding 641 amino acids. scVasa-s lacks a part of the N-terminal region found in the scVasa.Both of the two deduced amino acid sequences contain all of the known characteristics belonging to the DEAD-box protein family, four arginine-glycine-glycine(RGG) motifs and other eight conserved motifs.They show high similarity to Vasa homolog of Giebel carp(75.2%and 73.8%).Tissue distribution analysis by RT-PCR revealed that these two isoforms were exclusively expressed in the gonads of both female and male.In adult fish,scVasa was found to be mainly expressed in the primary oocytes atⅠandⅡphases in the ovary while in the spermatogonia and primary spermatocytes in the testis by in situ hybridization.Semi-quantitative RT-PCR analysis showed that the expressions of both scVasa isoforms were much higher in ovarian recrudescent stage(mainly with phaseⅡoocytes) than in vitellogenic stage(mainly with phaseⅢandⅣoocytes).
     This is the first time to clone so many germ cell markers in teleosts,the Nile tilapia and Southern catfish,both of which are important for aquaculture.Although investigations on these markers have been taken by reseachers all over the world extensively,most studies were focused only on a few model species,such as mouse,zebrafish and medaka,while little attentions have been paid to the important aquacultural speices.In order to lay foundations for growing monosex progeny and improving the growth rate of the aquacultural fishes by biologcial techeniques,expression patterns and changes during sex reversal by drug treatment of these germ cells and meiosis markers were investigated in the Nile tilapia and Southern catfish,two important aquacultural fish species in our country.The results of the study are important not only for researches on the mechanisms of sex differentiation,meiosis regulation and the cross talk between germ cells and somatic cells,but also important for practical implications in future aquaculture.
引文
[1]王德寿,吴天利.鱼类性别决定及其机制的研究进展[J].西南师范大学学报:自然科学版.2000,25(3):296-304.
    [2]Short R V.Difference between a testis and an ovary[M].1998:359-361.
    [3]Polanco J C,Koopman P.Sty and the hesitant beginnings of male development[J].Dev Biol.2007,302(1):13-24.
    [4]Wilhelm D,Palmer S,Koopman P.Sex determination and gonadal development in mammals[J].Physiol Rev.2007,87(1):1-28.
    [5]Born G G,Bertollo L A.An XX/XY sex chromosome system in a fish species Hoplias malabaricus,with a polymorpshic NOR bearing X chromosome.[J].Chromosome Res.2000(8):111-118.
    [6]Almeida T L,Daniel M F,Lopes C E.Sex chromosome evolution in fish Ⅱ.Second occurrence of an X1X2Y sex chromosome system in Gymnotiformes.[J].Chromosome Res.2000(8):335-340.
    [7]Ueno K,Ota K,Kobayashi T.Heteromorphic sex chromosomes of lizardfish(Synodontidae):focus on the ZZ-ZW1W2 system in Trachinocephalus myops.[J].Genetica.2001(111):133-142.
    [8]Artoni R F,Falcao J N,Moreira-filho.An uncommoncondition for a sex chromosome system in Characcidae fish.Distribution and differentiation of the ZZ/ZW system in Tripportheus.[J].Chromosome Res.2001(9):449-456.
    [9]Artoni R F,Bertollo L A.Trends in the karyotype evolution of Loricariidae fish(Siluriformes).[J].Hereditas.2001(134):201-210.
    [10]Jorge L C,Moreira-filho.Cytogenetic studies on Apareiodon affinis(Pisces,Characiformes) from Parana River basin:sex chromosomes and polymorphism.Genetica.[J].Genetica.2000(109):267-273.
    [11]Traut W,Winking H.Meiotic chromosomes and stages of sex chromosome evolution in fish:zebrafish,platyfish and guppy[J].Chromosome Res.2001,9(8):659-672.
    [12]Clemens H P,Inslee T.The production of unisex broods by Tilapia mossambica sex-reversed with methyltestosterone..[J].Transactions of the American Fisheries Society.1968(97):18-21.
    [13]Chang Z J,Yu Q X.The cytogenetic evidence of ZZ/ZW sex determinationin Paramisgumus dabryanus.[J].Hereditas.1997(19):17-19.
    [14]楼允东.鱼类育种学[M].北京:中国农业出版社,1999:196-201.
    [15]Caputo V M.Cytogenetics of nine species of Mediterranean blennies and additional evidence for anunusal Multiple sex-chromosome system in Parablennius tentacularis(Perciformes,Blenniidae).[J].Chromosome Res.2001(9):3-12.
    [16]Guan G,Kobayashi T,Nagahama Y.Sexually dimorphic expression of two types of DM(Doublesex/Mab-3)-domain genes in a teleost fish,the Tilapia(Oreochromis niloticus)[J].Biochem. Biophys.Res.Commun.2000,272(3):662-666.
    [17]Marchand Q,Govoroun M,Cotta D H.DMRTI expression during gonadal diferentiation and spermatogenesis in the rainbow trout,Oncorhynchus mykiss[J].Biochem.Biophys.2000,1493(1):180-187.
    [18]Wang D S,Kobayashi T,Senthilkumaran B.Molecular cloning of DAXI and SHP cDNAs and their expression patterns in the nile tilapia,Oreochromis niloticus[J].Biochem.Biophys.2002,297(3):632-640.
    [19]Yoshiuray,Senthikumaran B,Watanabe M,et al.Synergistic expression of Ad4BP/SF1 and cytochrome P450aromatase(ovarian type) in the ovary of Nile flapia,Oreochromis niloticus,during viteilogenesis suggests transcriptional interaction.[J].Biol.Reprod.2003,68(5):1545-1553.
    [20]Zhou R,Liu L,Guoy.Similar gene structure of two SOX9 a genes and their expression paterns during gonadal diferentiation in a teleost fish,rice field eel(Monopterus albus).[J].Mol,Reprod Dev.2003,66(3):211-217.
    [21]Wang D S,Kobayashi T,Zhou L Y.Molecular cloning and gene expression of FOX12 in the Nile tilapia,Oreochromis niloticus[J].Biochemical and Biophysical Research Communications.2004(320):83-89.
    [22]王德寿,吴天利.鱼类性别决定及其机制的研究进展[J].西南师范大学学报:自然科学版.2000,25(3):296-304.
    [23]周林燕,张修月,王德寿.脊椎动物性别决定和分化的分子机制研究进展[J].动物学研究.2004,25(1):81-88.
    [24]Kawano K,Furusawa S,Matsuda H,et al.Expression of steroidogenic factor-1 in frog embryo and developing gonad[M].2001:13-22.
    [25]Conover D O.Environmental sex determination interaction of temperature and gynotype in a fish[J].Science.1981(231):577-579.
    [26]Strussmann C A,Saito T,Usui M.Thermal thresholds and critical period of themiolabile sex determination in two atherinid fishes,Odonresthes bonariensis and Palogonina archery[J].J.Exp.Biol.1997(278):167-177.
    [27]Bogart M H.Sex determination:a hypothesis based on steroid ratios[J].J.Theor.Biol.1987(128):344-357.
    [28]Nagahama Y.Gonadal.steroid hormones:major regulators of gonadal sex differentiation and gametogenesis in fish.[J].In Reproductive Physiology of fish,.2000(278):211-222.
    [29]Devlin H R,Yoshitaka N.Gonadal.steroid hormones:major regulators of gonadal sex differentiation and gametogenesis in fish.[J].Aquaculture.2002(208):191-264.
    [30]Piferrer F,Baker I J.Effects of natural,synthetic,aromatizable,and nonaromatizablc androgens in inducing male sex differentiation in genotypic female chinook salmon(Oncorhynchus tshawytscha)[J].Gen Comp Endocrinol.1993,91(1):59-65,
    [31]Kavumpurath S,Pandian T J.Masculinization of fighting fish,Betta splendens Regan,using synthetic or natural androgens[J].Aquaculture Fish Manage.1994(25):373-381.
    [32]Olito C,Brock I.Sex reversal of rainbow trout creating an all-female population[J].Prog.Fish-culture.1991(53):41-44.
    [33]Goudie C A,Redner B D,Simco B A.Femnization of charnel catfish by oral administration of steroid sex hormones[J].Trans.Am.Fish.Soc.1983(112):670-667.
    [34]Solar L L.Optimalization of treatment regimes for sex controlled diferentiation and sterilization in wild rainbow trout by Oral administration of 17-amethyltestosterone[J].Aquaculture.1984(42):129-139.
    [35]Hurk R,Van C J,J J D.Effect of 17 alpha methyltestosterone and 11 beta hydroxyandrostenedione on gonad differentiation in the African catfish,Clarias gariepinus[J].Aquaculture.1989(83):179-191.
    [36]Yamamoto T.Sex differentiation[M].Academic Press,1969:117-175.
    [37]Guiguen Y J,Katia I O.Involvement of estrogens in the process of sex differentiation in two fish species:the rainbow trout(Oncorhynchus mykiss) and a tilapia(Oreochromis niloticus)[J].Molecular Reproduction and Development.1999(54):154-162.
    [38]Trant J M,Lehrter J,Gregory T,et al.Expression of cytochrome P450 aromatase in the channel catfish,Ictalurus punctatus[J].J Steroid Biochem Mol Biol.1997,61(3):393-397.
    [39]Smith C A,Katz M,Sinclair A H.DMRT1 Is Upregulated in the Gonads During Female-to-Male Sex Reversal in ZW Chicken Embryos[J].Biol Reprod.2003,68(2):560-570.
    [40]Davis K B.Hormonal sex manipulation and evidence for female homogamety in channel catfish[JJ.Gen.Comp.Endocrinol.1990(78):218-223.
    [41]D C H,Alexis F,Yann G,et al.Search for Genes Involved in the Temperature-Induced Gonadal Sex Differentiation in the Tilapia,Oreochromis niloticus[J].The Journal of experimental zoology.2001(290):574-585.
    [42]Liu S,Marina G,Helena D C.Expression of cytochrome P45011b(11b-hydroxylase) gene during gonadal sex differentiation and spermatogenesis in rainbow trout,Oncorhynchus mykiss[J].Journal of Steroid Biochemistry &Molecular Biology.2000(75):291-298.
    [43]Wibbels T,Bull J J,Crews D.Synergism between temperature and estradiol:a common pathway in turtle sex detennination[J].J.Exp.Zool.1999,260(1):130-134.
    [44]Kitano T,Takamune K K.Suppression of P450 aromatase gene expression in sex-reversed males produced by rearing genetically female larvae at high water temperature during a period of sex differentiation in the Japanese flounder(Paralichthys olivaceus)[J].Mo Endocrinol.1999,23(2):167-176.
    [45]Rm彼尔索夫.鱼类的性别分化[M].北京:农业出版社,1982:1-58.
    [46]刘筠.中国养殖鱼类繁殖生理学[M].北京:中国农业出版社,1993:1-201.
    [47]林光华,熊敬维.革胡子鲇卵巢在第一次性周期内分化与发育的研究[J].动物学研究.1995,16(4):365-372.
    [48]刘少军.革胡子鲇原始生殖细胞的起源,迁移及性腺分化[J].水生生物学报.1991,15(1):1-7.
    [49]高书堂.泥鳅原始生殖细胞的发生,迁移和性腺分化[J].武汉大学学报:自然科学版.1998,44(4):477-480
    [50]侯陵,刘文芳.温度对爬行动物性别的影响[J].生物学通报.1985(11):13-17.
    [51]Hamaguchi S.A light-and electron-microscopic study on the migration of primordial germ cells in the teleost,Oryzias latipes[J].Cell Tissue Res.1982,227(1):139-151.
    [52]Hogan J C.An ultrastructural analysis of “cytoplasmic markers” in germ cells of Oryzias latipes[J].J Ultrastruct Res.1978,62(3):237-250.
    [53]Braat A K,Zandbergen T,Van W S,et al.Characterization of zebrafish primordial germ ceils:morphology and early distribution of vasa RNA[J].Dev Dyn.1999,216(2):153-167.
    [54]Knaut H,Pelegri F,Bohmann K,et al.Zebrafish vasa RNA but not its protein is a component of the germ plasm and segregates asymmetrically before germline specification[J].J Cell Biol.2000,149(4):875-888.
    [55]Yoon C,Kawakami K,Hopkins N.Zebrafish vasa homologue RNA is localized to the cleavage planes of 2-and 4-cell-stage embryos and is expressed in the primordial germ ceils[J].Development.1997,124(16):3157-3165.
    [56]Satoh N.An ultrastructural study of sex differentiation in the teleost Oryzias latipes[J].J Embryol Exp Morphol.1974,32(1):195-215.
    [57]Billard R.Ultrastructural changes in the spermatogonia and spermatocytes of Poecilia reticulata during spermatogenesis[J].Cell Tissue Res.1984,237(2):219-226.
    [58]Hardisty M W.The numbers of vertebrate primordial germ cells[J].Biol Rev Camb Philos Soc.1967,42(2):265-287.
    [59]Nedelea M,Steopoe I.[Origin,cytologic characteristics and behavior of primary gonocytes during embryogenesis in the young larvae of Cyprinus carpio L.(Teleosteans)][J].Anat Ariz.1970,127(3):338-346.
    [60]Hardisty C A,Munro D S.Advances in endocrinology[J].Practitioner.1978,221(1324):499-503.
    [61]Pala M.The embryonic history of the primordial germ cells in Gambusia holbrookii(Grd).[J].Bull.zool.1970(37):49-62.
    [62]Gamo H.On the origin of germ cells and formation of gonad primordial in the medaka Orycias latipes.[J].Jap.J.zool.1961(13):101-105.
    [63]王国恩.鱼类的原生殖细胞[J].大连水产学院学报.1987(02):19-26.
    [64]Bowles J,Knight D,Smith C,et al.Retinoid signaling determines germ cell fate in mice[J].Science.2006,312(5773):596-600.
    [65]Atz J W.lntersexuality in fishes[M].Academic Press,London,1964:145-232.
    [66]Lin E,Dabrowski K,Timmermans L P,et al.Early gonadal development and sexual diferentiation in muskellunge(Esox masquinongy)[J].J.Zool.1997(75):1262-1269.
    [67]Takahashi H.Juvenile hermaphroditism in the zebrafish,Brachydanio rerio.[J].Bull.Fac Fish.Hokkaido Univ.1977(8):57-65.
    [68]Takahashi H,Shimizu M.JJuvenile intersexuality in a cyprinid fish,the Sumatra barb Barbus tetrazona.[J]. Bull.Fac Fish.Hokkaido Univ.1977(34):69-78.
    [69]Saitou M B,Surani M A.A molecular programme for the specification of germ cell fate in mice diferentiation in muskellunge(Esox masquinongy)[J].Nature.2002(418):293-300.
    [70]Eiges R,Benvenisty N.A molecular view on pluripotent stem cells[J].FEBS Lett.2002,529(1):135-141.
    [71]Nichols J,Zevnik B,Anastassiadis K,et al.Formation of pluripotent stem cells in the mammalian embryo depends on the POU transcription factor Oct4[J].Cell.1998,95(3):379-391.
    [72]Rajpert-de M E,Hanstein R,Jorgensen N,et al.Developmental expression of POUSF1(OCT-3/4) in normal and dysgenetic human gonads[J].Human Reproduction.2004,19(6):1338-1344.
    [73]Chambers I,Colby D,Robertson M,et al.Functional expression cloning of Nanog,a pluripotency sustaining factor in embryonic stem cells[J].Ceil.2003,113(5):643-655.
    [74]Chuma S,Kanatsu-shinohara M,Inoue K,et al.Spermatogenesis from epiblast and primordial germ cells following transplantation into postnatal mouse testis[J].Development.2005,132(1):117-122.
    [75]Mitsui K,Yokuzawa Y,ltoh H,et al.The homeoprotein Nanog is required for maintenance of pluripotency in mouse epiblast and ES cells[J].Cell.2003,113(5):631-642.
    [76]Obata Y,Kono T,Hatada I.Gene silencing:maturation of mouse fetal germ cells in vitro[J].Nature.2002,418(6897):497.
    [77]Zeineddine D,Papadimou E,Chebli K,et al.Oct-3/4 dose dependently regulates specification of embryonic stem cells toward a cardiac lineage and early heart development[J].Developmental Cell.2006,11(4):535-546.
    [78]Dickstein K,De V H,Miric M P,et al.Effect of single doses of SLV306,an inhibitor of both neutral endopeptidase and endothelin-converting enzyme,on pulmonary pressures in congestive heart failure[J].Am J Cardiol.2004,94(2):237-239.
    [79]Clark A T,Bodnar M S,Fox M,et al.Spontaneous differentiation of germ cells from human embryonic stem cells in vitro[J].Hum Mol Genet.2004,13(7):727-739.
    [80]Zhao G Q,Gathers D L.Male germ cell specification and differentiation[J].Dev Cell.2002,2(5):537-547.
    [81]Lacham-kaplan O,Chy H,Trounson A.Testicular cell conditioned medium supports differentiation of embryonic stem cells into ovarian structures containing oocytes[J].Stem Ceils.2006,24(2):266-273.
    [82]戈贤平,刘柱军.淡水优质鱼类养殖大全[M].2005:415-436.
    [83]Hines G A,Boots L R,Wibbels T,et al.Steroid levels and steroid metabolism in relation to early gonadal development in the tilapia Oreochromis niloticus(Teleostei:cyprinoidei)[J].Gen Comp Endocrinol.1999,114(2):235-248.
    [84]林浩然.鱼类生理学[M].广州:广东高等教育出版社,1999:193-201.
    [85]Schu P B.Maternal-effect mutations altering the anterior-posterior pattern of the Drosophila embryo[J].Roux's Arch.Dev.Biol.1986,195:302-317.
    [86]Gruidl M E,Smith P A,Kuznicki K A,et al.Multiple potential germ-line helicases are components of the germ-line-specific P granules of Caenorhabditis elegans[J].Proc Natl Acad Sci U S A.1996, 93(24):13837-13842.
    [87]Nakao H.Isolation and characterization of a Bombyx vasa-like gene[J].Dev Genes Evol.1999, 209(5):312-316.
    [88]Fabioux C, Huvet A, Lelong C, et al.Oyster vasa-like gene as a marker of the germline cell development in Crassostrea gigas[J], BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS.2004, 320(2):592-598.
    [89]Fabioux C, Pouvreau S, Le R F, et al.The oyster vasa-like gene: a specific marker of the germline in Crassostrea gigas[J].BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS.2004, 315(4):897-904.
    [90]Olsen L C, Aasland R, Fjose A.A vasa-like gene in zebrafish identifies putative primordial germ cells[J].Mech Dev.1997, 66(1-2): 95-105.
    [91]Kobayashi T, Kajiura-kobayashi H, Nagahama Y.Differential expression of vasa homologue gene in the germ cells during oogenesis and spermatogenesis in a teleost fish, tilapia, Oreochromis niloticus[J].Mech Dev.2000,99(1-2): 139-142.
    [92]Komiya T, Itoh K, Ikenishi K, et al.Isolation and characterization of a novel gene of the DEAD box protein family which is specifically expressed in germ cells of Xenopus laevis[J].Dev Biol.1994, 162(2): 354-363.
    [93]Tsunekawa N, Naito M, Sakai Y, et al.Isolation of chicken vasa homolog gene and tracing the origin of primordial germcells[J].Development.2000,127(12): 2741-2750.
    [94]Abe K, Noce T.A DEAD-family protein gene, Ddx4, encoding a murine homolog of Drosophila vasa maps to the distal end of mouse chromosome 13[J].Mamm Genome.1997, 8(8): 622-623.
    [95]Castrillon D H, Quade B J, Wang T Y, et al.The human VASA gene is specifically expressed in the germ cell lineage[J].Proc Natl Acad Sci U S A.2000, 97(17): 9585-9590.
    [96]Mochizuki K, Nishimiya-fujisawa C, Fujisawa T.Universal occurrence of the vasa-related genes among metazoansand their germline expression in Hydra[J].Dev Genes Evol.2001, 211(6): 299-308.
    [97]Extavour C G, Akam M.Mechanisms of germ cell specification across the metazoans: epigenesis and preformation [J].Development.2003, 130(24): 5869-5884.
    [98]Extavour C G, Pang K, Matus D Q, et al.vasa and nanos expression patterns in a sea anemone and the evolution of bilaterian germ cell specification mechanisms[J].EVOLUTION & DEVELOPMENT.2005, 7(3):201-215.
    [99]Yoshizaki G, Takeuchi Y, Sakatani S, et al.Germ cell-specific expression of green fluorescent protein in transgenic rainbow trout under control of the rainbow trout vasa-like gene promoter[J].Int J Dev Biol.2000, 44(3):323-326.
    [100]Krovel A V, Olsen L C.Expression of a vas::EGFP transgene in primordial germ cells of the zebrafish[J]. Mech Dev.2002,116(1-2):141-150.
    [101]Tanaka M,Kinoshita M,Kobayashi D,et al.Establishment of medaka(Oryzias latipes) transgenic lines with the expression of green fluorescent protein fluorescence exclusively in germ cells:a useful model to monitor germ cells in a live vertebrate[J].Proc Natl Acad Sci U S A.2001,98(5):2544-2549.
    [102]Yano A,Suzuki K,Yoshizaki G.Flow-cytometric isolation of testicular germ cells from rainbow trout (Oncorhynchus mykiss) carrying the green fluorescent protein gene driven by trout vasa regulatory regions[J].Biol Reprod.2008,78(1):151-158.
    [103]Takeuchi Y,Yoshizaki G,Kobayashi T,et al.Mass isolation of primordial germ cells from transgenic rainbow trout carrying the green fluorescent protein gene driven by the vasa gene promoter[J].Biol Reprod.2002,67(4):1087-1092.
    [104]Ye D,Lv D,Song P,et al.Cloning and characterization of a rice field eel vasa-like gene eDNA and its expression in gonads during natural sex transformation[J].Biochem Genet.2007,45(3-4):211-224.
    [105]Xu H,Gui J,Hong Y.Differential expression of vasa RNA and protein during spermatogenesis and oogenesis in the gibel carp(Carassius auratus gibelio),a bisexually and gynogenetically reproducing vertebrate[J].Dev Dyn.2005,233(3):872-882.
    [106]陈云贵,叶鼎,宋平,等.金鱼配子发生中vasa基因的表达和分布特征[J].动物学研究.2005,26(2):179-183.
    [107]张耀光,谢小军.南方鲇的繁殖生物学研究:性腺发育及周年变化[J].水生生物学报.1996,20(1):8-16
    [108]Liu Z,Wu F,Jiao B,et al.Molecular cloning of doublesex and mab-3-related transcription factor 1,forkhead transcription factor gene 2,and two types of cytochrome P450 aromatase in Southern catfish and their possible roles in sex differentiation[J].J Endocrinol.2007,194(1):223-241.
    [109]1996(01).(In Chinese)
    [110]Kobayashi T,Kajiura-kobayashi H,Nagahama Y.Two isoforms of vasa homologs in a teleost fish:their differential expression during germ cell differentiation[J].Mech Dev.2002,111(1-2):167-171.
    [111]Pause A,Sonenberg N.Mutational analysis of a DEAD box RNA helicase:the mammalian translation initiation factor eIF-4A[J].EMBO J.1992,11(7):2643-2654.
    [112]Kiledjian M,Dreyfuss G.Primary structure and binding activity of the hnRNP U protein:binding RNA through RGG box[J].EMBO J.1992,11(7):2655-2664.
    [113]Fujiwara Y,Komiya T,Kawabata H,et al.Isolation of a DEAD-family protein gene that encodes a murine homolog of Drosophila vasa and its specific expression in germ cell lineage[J].Proc Natl Acad Sci U S A.1994,91(25):12258-12262.
    [114]Tanaka S S,Toyooka Y,Akasu R,et al.The mouse homolog of Drosophila Vasa is required for the development of male germ cells[J].Genes Dev.2000,14(7):841-853.
    [115]Zeeman A M,Stoop H,Boter M,et al.VASA is a specific marker for both normal and malignant human germ cells[J].Lab Invest.2002, 82(2): 159-166.
    [116]张红卫,王子仁,张士璀.发育生物学.[M].北京:高等教育出版社,2006:399-407.
    [117]Howley C, Ho R K.mRNA localization patterns in zebrafish oocytes[J].Mech Dev.2000,92(2): 305-309.[118]1996(01).(In Chinese)
    [119]Venables J P, Ruggiu M, Cooke H J.The RNA-binding specificity of the mouse Dazl protein.[J].Nucleic Acids Res.2001,29(12): 2479-2483.
    [120]Van G R, Tuerlings J H, Kremer J A, et ai.DAZLA: an important candidate gene in male subfertility?[J].J Assist Reprod Genet.2001, 18(7): 395-399.
    [121]Yen P H.Putative biological functions of the DAZ family.[J].Int J Androl.2004,27(3): 125-129.
    [122]Tung J Y, Rosen M P, Nelson L M, et al.Variants in Deleted in AZoospermia-Like (DAZL) are correlated with reproductive parameters in men and women.[J], Hum Genet.2006,118(6): 730-740.
    [123]Tung J Y, Rosen M P, Nelson L M, et al.Novel missense mutations of the Deleted-in-AZoospermia-Like (DAZL) gene in infertile women and men.[J].Reprod Biol Endocrinol.2006,4: 40.
    [124]Dobson M J, Pearlman R E, Karaiskakis A, et al.Synaptonemal complex proteins: occurrence, epitope mapping and chromosome disjunction[J].J Cell Sci.1994, 107 (Pt 10): 2749-2760.
    [125]Fox M, Urano J, Reijo P R.Identification and characterization of RNA sequences to which human PUMILIO-2 (PUM2) and deleted in Azoospermia-like (DAZL) bind.[J].Genomics.2005, 85(1): 92-105.
    [126]Kuo P L, Wang S T, Lin Y M, et al.Expression profiles of the DAZ gene family in human testis with and without spermatogenic failure.[J].Fertil Steril.2004, 81(4): 1034-1040.
    [127]Schrans-stassen B H, Saunders P T, Cooke H J, et al.Nature of the spermatogenic arrest in Dazl -/- mice.[J].Biol Reprod.2001,65(3): 771-776.
    [128]Dai T, Vera Y, Salido E C, et al.Characterization of the mouse Dazapl gene encoding an RNA-binding protein that interacts with infertility factors DAZ and DAZL.[J].BMC Genomics.2001, 2(1): 6.
    [129]Kosaka K, Kawakami K, Sakamoto H, et al.Spatiotemporal localization of germ plasm RNAs during zebrafish oogenesis.[J].Mech Dev.2007, 124(4): 279-289.
    [130]Schalk J A, Dietrich A J, Vink A C, et al.Localization of SCP2 and SCP3 protein molecules within synaptonemal complexes of the rat[J].Chromosoma.1998,107(8): 540-548.
    [131]Yuan L, Liu J G, Zhao J, et al.The murine SCP3 gene is required for synaptonemal complex assembly ,chromosome synapsis, and male fertility[J].Mol Cell.2000,5(1): 73-83.
    [132]Grossmann B, Weinbauer G, Hirschmann P, et al.Conservation of the deleted-in-azoospermia-like-1 (DAZL1) gene structure in old world monkeys points to a homologous function of DAZL1 in this primate class.[J].J Endocrinol Invest.2000, 23(9): 616-622.
    [133]Houston D W, King M L.A critical role for Xdazl, a germ plasm-localized RNA, in the differentiation of primordial germ cells in Xenopus[J].Development.2000, 127(3): 447-456.
    [134]Lee J H, Lee D R, Yoon S J, et al.Expression of DAZ (deleted in azoospermia), DAZL1 (DAZ-like) and protamine-2 in testis and its application for diagnosis of spermatogenesis in non-obstructive azoospermia.[J].Mol Hum Reprod.1998,4(9): 827-834.
    [135]Tsui S, Dai T, Roettger S, et al.Identification of two novel proteins that interact with germ-cell-specific RNA-binding proteins DAZ and DAZLI.[J].Genomics.2000, 65(3): 266-273.
    [136]Reynolds N, Collier B, Bingham V, et al.Translation of the synaptonemai complex component Sycp3 is enhanced in vivo by the germ cell specific regulator Dazl.[J].RNA.2007,13(7): 974-981.
    [137]Yuan L, Pelttari J, Brundell E, et al.The synaptonemai complex protein SCP3 can form multistranded, cross-striated fibers in vivo[J].J Cell Biol.1998,142(2): 331-339.
    [138]Baier A, Alsheimer M, Benavente R.Synaptonemai complex protein SYCP3: Conserved polymerization properties among vertebrates[J].Biochim Biophys Acta.2007, 1774(5): 595-602.
    [139]Yamamoto Y, Usui F, Nakamura Y, et al.A novel method to isolate primordial germ cells and its use for the generation of germline chimeras in chicken.[J].Biol Reprod.2007, 77(1): 115-119.
    [140]Yuan L, Pelttari J, Brundell E, et al.The synaptonemai complex protein SCP3 can form multistranded, cross-striated fibers in vivo[J].J Cell Biol.1998, 142(2): 331-339.
    [141]Lucas H, Patrat C, Jouannet P, et al.A novel, rapid, and accurate method for detecting microdeletion involving the DAZ gene in infertile men.[J].Fertil Steril.2000, 73(2): 242-247.
    [142]Baier A, Alsheimer M, Benavente R.Synaptonemai complex protein SYCP3: Conserved polymerization properties among vertebrates[J].Biochim Biophys Acta.2007,1774(5): 595-602.
    [143]De R D.Proliferation and differentiation of spermatogonia! stem cells.[J].Reproduction.2001, 121(3):347-354.
    [144]Baier A, Alsheimer M, Volff J -, et al.Synaptonemai Complex Protein SYCP3 of the Rat: Evolutionarily Conserved Domains and the Assembly of Higher Order Structures[J].Sexual Development Sex Dev.2007, 1(3): 161-168.
    [145]Wang P J, Mccarrey J R, Yang F, et al.An abundance of X-linked genes expressed in spermatogonia[J].Nat Genet.2001, 27(4): 422-426.
    [146]Xu H, Li M, Gui J, et al.Cloning and expression of medaka dazl during embryogenesis and gametogenesis.[J].Gene Expr Patterns.2007, 7(3): 332-338.
    [147]Carr-schmid A, Jiao X, Kiledjian M.Identification of mRNA bound to RNA binding proteins by differential display[J].Methods Mol Biol.2006, 317: 299-314.
    [148]Dobson M J, Pearlman R E, Karaiskakis A, et al.Synaptonemai complex proteins: occurrence, epitope mapping and chromosome disjunction[J], J Cell Sci.1994, 107 ( Pt 10): 2749-2760.
    [149]Fox M, Urano J, Reijo P R.Identification and characterization of RNA sequences to which human PUMILIO-2 (PUM2) and deleted in Azoospermia-like (DAZL) bind[J], Genomics.2005, 85(1): 92-105.
    [150]Yuan L, Liu J G, Hoja M R, et al.Female germ cell aneuploidy and embryo death in mice lacking the meiosis-specific protein SCP3[J].Science.2002, 296(5570): 1115-1118.
    [151]Bin L, Gang W, Hu J, et al.Identification and characterization of TSAP, a novel gene specifically expressed in testis during spermatogenesis.[J].Mol Reprod Dev.2007, 74(9): 1141-1148.
    [152]Lin Y, Page D C.Dazl deficiency leads to embryonic arrest of germ cell development in XY C57BL/6 mice.[J].Dev Biol.2005,288(2): 309-316.
    [153]Yuan L, Liu J G, Zhao J, et al.The murine SCP3 gene is required for synaptonemal complex assembly,chromosome synapsis, and male fertility[J].Mol Cell.2000,5(1): 73-83.
    [154]Carr-schmid A, Jiao X, Kiledjian M.Identification of mRNA bound to RNA binding proteins by differential display.[J], Methods Mol Biol.2006, 317: 299-314.
    [155]Yano A, Suzuki K, Yoshizaki G.Flow-Cytometric Isolation of Testicular Germ Cells from Rainbow Trout (Oncorhynchus mykiss) Carrying the Green Fluorescent Protein Gene Driven by Trout vasa Regulatory Regions[Tj.Biol Reprod.2007: biolreprod.107.064667.
    [156]Iwai T, Lee J, Yoshii A, et al.Changes in the expression and localization of cohesin subunits during meiosis in a non-mammalian vertebrate, the medaka fish[J].Gene Expr Patterns.2004,4(5): 495-504.
    [157]Iwai T, Yoshii A, Yokota T, et al.Structural components of the synaptonemal complex, SYCP1 and SYCP3, in the medaka fish Oryzias latipes[J].Exp Cell Res.2006,312(13): 2528-2537.
    [158]Reynolds N, Collier B, Maratou K, et al.Dazl binds in vivo to specific transcripts and can regulate the pre-meiotic translation of Mvh in germ cells[J].Hum Mol Genet.2005, 14(24): 3899-3909.
    [159]Reynolds N, Collier B, Bingham V, et al.Translation of the synaptonemal complex component Sycp3 is enhanced in vivo by the germ cell specific regulator DazI[J].RNA.2007, 13(7): 974-981.
    [160]Bally-cuif L, Schatz W J, Ho R K.Characterization of the zebrafish Orb/CPEB-related RNA binding protein and localization of maternal components in the zebrafish oocyte.[J].Mech Dev.1998, 77(1): 31-47.
    [161]Lucas H, Patrat C, Jouannet P, et al.A novel, rapid, and accurate method for detecting microdeletion involving the DAZ gene in infertile men[J].Fertil Steril.2000,73(2): 242-247.
    [162]Yamamoto Y, Usui F, Nakamura Y, et al.A novel method to isolate primordial germ cells and its use for the generation of germline chimeras in chicken[J].Biol Reprod.2007, 77(1): 115-119.
    [163]Baier A, Aisheimer M, Volff J -, et al.Synaptonemal Complex Protein SYCP3 of the Rat: Evolutionanly Conserved Domains and the Assembly of Higher Order Structures[J].Sexual Development Sex Dev.2007, 1(3): 161-168.
    [164]De R D.Proliferation and differentiation of spermatogonial stem cells[J].Reproduction.2001, 121(3): 347-354.
    [165]Yano A, Suzuki K, Yoshizaki G.Flow-Cytometric Isolation of Testicular Germ Cells from Rainbow Trout (Oncorhynchus mykiss) Carrying the Green Fluorescent Protein Gene Driven by Trout vasa Regulatory Regions[J].Biol Reprod.2007: biolreprod.107.064667.
    [166]Schalk J A,Dietrich A J,Vink A C,et al.Localization of SCP2 and SCP3 protein molecules within synaptonemal complexes of the rat[J].Chromosoma.1998,107(8):540-548.
    [167]Grossmann B,Weinbauer G,Hirschmann P,et al.Conservation of the deleted-in-azoospermia-like-1(DAZLI)gene structure in old world monkeys points to a homologous function of DAZL1 in this primate class[J].J Endocrinol Invest.2000,23(9):616-622.
    [168]Reynolds N,Collier B,Bingham V,et al.Translation of the synaptonemal complex component Sycp3 is enhanced in vivo by the germ cell specific regulator Dazl[J].RNA.2007,13(7):974-981.
    [169]Eijpe M,Offenberg H,Jessberger R,et al.Meiotic cohesin REC8 marks the axial elements of rat synaptonemal complexes before cohesins SMC1 beta and SMC3.[J].J Cell Biol.2003,160(5):657-670.
    [170]Lee J,Iwai T,Yokota T,et al.Temporally and spatially selective loss of Rec8 protein from meiotic chromosomes during mammalian meiosis.[J].J Cell Sci.2003,116(Pt 13):2781-2790.
    [171]Soyal S M,Amleh A,Dean J.FIGalpha,a germ cell-specific transcription factor required for ovarian follicle formation[J].Development.2000,127(21):4645-4654.
    [172]黄健,郑月慧,徐良全.原始卵泡生长启动调控机制的研究进展[J].生殖医学杂志.2007,16(5):372-376.
    [173]Bayne R A,Martins S S,Anderson R A.Increased expression of the FIGLA transcription factor is associated with primordial follicle formation in the human fetal ovary[J].Mol Hum Reprod.2004,10(6):373-381.
    [174]Onichtchouk D,Aduroja K,Belting H G,et al.Transgene driving GFP expression from the promoter of the zona pellucida gene zpc is expressed in oocytes and provides an early marker for gonad differentiation in zebrafish[J].Dev Dyn.2003,228(3):393-404.
    [175]Kanamori A,Yamamura A,Koshiba S,et al.Methyltestosterone efficiently induces male development in the self-fertilizing hermaphrodite fish,Kryptolebias marmoratus[M].2006:495-503.
    [176]Liang L,Soyal S M,Dean J.FIGalpha,a germ cell specific transcription factor involved in the coordinate expression of the zona pellucida genes[J].Development.1997,124(24):4939-4947.
    [177]李建义,童坦君.螺旋—环区—螺旋蛋白质—DNA结合蛋白的新类型[J].生物化学与生物物理进展.1993,20(1):11-14.
    [178]贺强,王立峰.bHLH蛋白家族的功能[J].国外医学:生理病理科学与临床分册.2004,24(6):545-547.
    [179]章为.组织学与胚胎学[M].四川大学出版社,2005:227-230.
    [180]陈诗书,汤雪明.医学细胞与分子生物学(第二版)[M].科学出版社,2003:651-660.
    [181]Zamboni L,Upadhyay S.Germ cell differentiation in mouse adrenal glands[J].J Exp Zool.1983,228(2):173-193.
    [182]Joshi N S,Cui W,Chandele A,et al.Inflammation directs memory precursor and short-lived effector CD8(+) T cell fates via the graded expression of T-bet transcription factor[J].Immunity.2007,27(2):281-295.
    [183]Epifano O,Liang L F,Familari M,et al.Coordinate expression of the three zona pellucida genes during mouse oogenesis[J].Development.1995,121(7):1947-1956.
    [184]Miller D L, Papayannopoulos I A, Styles J, et al.Peptide compositions of the cerebrovascular and senile plaque core amyloid deposits of Alzheimer's disease[J].Arch Biochem Biophys.1993,301(1): 41-52.
    [185]Pesce M, Wang X, Wolgemuth D J, et al.Differential expression of the Oct-4 transcription factor during mouse germ cell differentiation[J].Mech Dev.1998, 71(1-2): 89-98.
    [186]Yoshiura Y, Senthilkumaran B, Watanabe M, et al.Synergistic expression of Ad4BP/SF-l and cytochrome P-450 aromatase (ovarian type) in the ovary of Nile tilapia, Oreochromis niloticus, during vitellogenesis suggests transcriptional interaction.[J].Biol Reprod.2003,68(5): 1545-1553.
    [187]Kobayashi T, Kajiura-kobayashi H, Nagahama Y.Differential expression of vasa homologue gene in the germ cells during oogenesis and spermatogenesis in a teleost fish, tilapia, Oreochromis niloticus[J].Mech Dev.2000,99(1-2): 139-142.
    [188]Huntriss J, Gosden R, Hinkins M, et al.Isolation, characterization and expression of the human Factor In the Germline alpha (FIGLA) gene in ovarian follicles and oocytes[J], Mol Hum Reprod.2002, 8(12): 1087-1095.
    [189]Kanamori A.Systematic identification of genes expressed during early oogenesis in medaka[J].Mol Reprod Dev.2000, 55(1): 31-36.
    [190]Kanamori A, Yamamura A, Koshiba S, et al.Methyltestosterone efficiently induces male development in the self-fertilizing hermaphrodite fish, Kryptolebias marmoratus[J].Genesis.2006,44(10): 495-503.
    [191]Siaud N, Dray E, Gy I, et al.Brca2 is involved in meiosis in Arabidopsis thaliana as suggested by its interaction with Dmcl [J].Embo J.2004, 23(6): 1392-1401.
    [192]Mclaren A.The fate of germ cells in the testis of fetal Sex-reversed mice[M].1981: 461-467.
    [193]Mclaren A.Primordial germ cells in the mouse[J].Dev Biol.2003, 262(1): 1-15.
    [194]Oliver B.Genetic control of germline sexual dimorphism in Drosophila[J].Int Rev Cytol.2002, 219: 1-60.
    [195]Steinmann-zwicky M, Schmid H, Nothiger R.Cell-autonomous and inductive signals can determine the sex of the germ line ofdrosophila by regulating the gene Sxl[J].Cell.1989, 57(1): 157-166.
    [196]Okamoto K, Okazawa H, Okuda A, et al.A novel octamer binding transcription factor is differentially expressed in mouse embryonic cells[J].Cell.1990, 60(3): 461-472.
    [197]Rosner M H, Vigano M A, Ozato K, et al.A POU-domain transcription factor in early stem cells and germ cells of the mammalian embryo[J].Nature.1990, 345(6277): 686-692.
    [198]Scholer H R, Dressier G R, Balling R, et al.Oct-4: a germline-specific transcription factor mapping to the mouse t-complex[J].Embo J.1990, 9(7): 2185-2195.
    [199]Takeda J, Seino S, Bell G I.Human Oct3 gene family: cDNA sequences, alternative splicing, gene organization,chromosomal location, and expression at low levels in adult tissues[J].Nucleic Acids Res.1992, 20(17):4613-4620.
    [200]Guillaudeux T, Mattei M G, Depetris D, et al.In situ hybridization localizes the human OTF3 to chromosome 6p21.3-->p22 and OTF3L to 12p13.[J].Cytogenet Cell Genet.1993, 63(4): 212-214.
    [201]Crouau-roy B, Amadou C, Bouissou C, et al.Localization of the OTF3 gene within the human MHC class Ⅰ region by physical and meiotic mapping.[J].Genomics.1994,21(1): 241-243.
    [202]Scholer H R, Dressier G R, Balling R, et al.Oct-4: a germline-specific transcription factor mapping to the mouse t-complex[J].Embo J.1990, 9(7): 2185-2195.
    [203]Yeom Y I, Fuhrmann G, Ovitt C E, et al.Germline regulatory element of Oct-4 specific for the totipotent cycle of embryonal cells[J].Development.1996,122(3): 881-894.
    [204]Rosner M H, Vigano M A, Ozato K, et al.A POU-domain transcription factor in early stem cells and germ cells of the mammalian embryo[J].Nature.1990,345(6277): 686-692.
    [205]Donovan P J, De M M.Turning germ cells into stem cells[M].2003: 463-471.
    [206]Surani M A.Reprogramming of genome function through epigenetic inheritance[J].Nature.2001, 414(6859):122-128.
    [207]Niwa H, Miyazaki J, Smith A G.Quantitative expression of Oct-3/4 defines differentiation, dedifferentiation or self-renewal of ES cells[J].Nature Genetics.2000,24(4): 372-376.
    [208]Tadokoro Y, Yomogida K, Ohta H, et al.Homeostatic regulation of germinal stem cell proliferation by the GDNF/FSH pathway[J].Mech Dev.2002,113(1): 29-39.
    [209]Wernig M, Meissner A, Foreman R, et al.In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state.[J].Nature.2007,448(7151): 318-324.
    [210]Takahashi K, Yamanaka S.Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors[J].Cell.2006, 126(4): 663-676.
    [211]Kehler J, Tolkunova E, Koschorz B, et al.Oct4 is required for primordial germ cell survival[J].EMBO Rep.2004,5(11): 1078-1083.
    [212]Agulnik A I, Zharkikh A, Boettger-tong H, et al.Evolution of the DAZ gene family suggests that Y-linked DAZ plays little, or a limited, role in spermatogenesis but underlines a recent African origin for human populations[J].Hum Mol Genet.1998,7(9): 1371-1377.
    [213]Nishimoto M, Fukushima A, Okuda A, et al.The gene for the embryonic stem cell coactivator UTF1 carries a regulatory element which selectively interacts with a complex composed of Oct-3/4 and Sox-2[J].Mol Cell Biol.1999, 19(8): 5453-5465.
    [214]Ben-shushan E, Thompson J R, Gudas L J, et al.Rex-1, a gene encoding a transcription factor expressed in the early embryo, is regulated via Oct-3/4 and Oct-6 binding to an octamer site and a novel protein, Rox-1, binding toan adjacent site[J].Mol Cell Biol.1998, 18(4): 1866-1878.
    [215]Tokuzawa Y, Kaiho E, Maruyama M, et al.Fbx15 is a novel target of Oct3/4 but is dispensable for embryonic stem cell self-renewal and mouse development[J].Mol Cell Biol.2003, 23(8): 2699-2708.
    [216]Kuroda T, Tada M, Kubota H, et al.Octamer and Sox elements are required for transcriptional cis regulation of Nanog gene expression[J], Mol Cell Biol.2005, 25(6): 2475-2485.
    [217]Rodda D J, Chew J L, Lim L H, et al.Transcriptional regulation of nanog by OCT4 and SOX2[J].J Biol Chem.2005,280(26): 24731-24737.
    [218]Tomioka M, Nishimoto M, Miyagi S, et al.Identification of Sox-2 regulatory region which is under the control of Oct-3/4-Sox-2 complex[J].Nucleic Acids Res.2002,30(14): 3202-3213.
    [219]Dailey L, Yuan H, Basilico C.Interaction between a novel F9-specific factor and octamer-binding proteins is required for cell-type-restricted activity of the fibroblast growth factor 4 enhancer.[J], Mol Cell Biol.1994, 14(12):7758-7769.
    [220]Ambrosetti D C, Basilico C, Dailey L.Synergistic activation of the fibroblast growth factor 4 enhancer by Sox2 and Oct-3 depends on protein-protein interactions facilitated by a specific spatial arrangement of factor binding sites[J].Mol Cell Biol.1997, 17(11): 6321-6329.
    [221]Barnea E, Bergman Y.Synergy of SF1 and RAR in activation of Oct-3/4 promoter[J].J Biol Chem.2000,275(9): 6608-6619.
    [222]Gu P, Goodwin B, Chung A C, et al.Orphan nuclear receptor LRH-1 is required to maintain Oct4 expression at the epiblast stage of embryonic development[J], Mol Cell Biol.2005, 25(9): 3492-3505.[223]Fuhrmann G, Chung A C, Jackson K J, et al.Mouse germline restriction of Oct4 expression by germ cell nuclear factor[J].Dev Cell.2001, 1(3): 377-387.
    [224]Gu P, Lemenuet D, Chung A C, et al.Orphan nuclear receptor GCNF is required for the repression of pluripotency genes during retinoic acid-induced embryonic stem cell differentiation.[J].Mol Cell Biol.2005,25(19): 8507-8519.
    [225]Ben-shushan E, Sharir H, Pikarsky E, et al.A dynamic balance between ARP-1/COUP-TFⅡ,EAR-3/COUP-TFI, and retinoic acid receptonretinoid X receptor heterodimers regulates Oct-3/4 expression in embryonal carcinoma cells[J].Mol Cell Biol.1995, 15(2): 1034-1048.
    [226]Pikarsky E, Sharir H, Ben-shushan E, et al.Retinoic acid represses Oct-3/4 gene expression through several retinoic acid-responsive elements located in the promoter-enhancer region[J].Mol Cell Biol.1994, 14(2):1026-1038.
    [227]Schoorlemmer J, Van P A, den Van E, et al.Characterization of a negative retinoic acid response element in the murine Oct4 promoter[J].Mol Cell Biol.1994, 14(2): 1122-1136.
    [228]Chew J L, Loh Y H, Zhang W, et al.Reciprocal transcriptional regulation of Pou5fl and Sox2 via the Oct4/Sox2 complex in embryonic stem cells[J].Mol Cell Biol.2005, 25(14): 6031-6046.
    [229]Okumura-nakanishi S, Saito M, Niwa H, et al.Oct-3/4 and Sox2 regulate Oct-3/4 gene in embryonic stem cells[J].J Biol Chem.2005, 280(7): 5307-5317.
    [230]Yeom Y I, Fuhrmann G, Ovitt C E, et al.Germline regulatory element of Oct-4 specific for the totipotent cycle of embryonal cells[J].Development.1996, 122(3): 881-894.
    [231]Nordhoff V, Hubner K, Bauer A, et al.Comparative analysis of human, bovine, and murine Oct-4 upstream promoter sequences[J].Mamm Genome.2001,12(4): 309-317.
    [232]Gu P, Goodwin B, Chung A C, et al.Orphan nuclear receptor LRH-1 is required to maintain Oct4 expression at the epiblast stage of embryonic development[J].Mol Cell Biol.2005,25(9): 3492-3505.
    [233]Guo Y, Costa R, Ramsey H, et al.The embryonic stem cell transcription factors Oct-4 and FoxD3 interact to regulate endodermal-specific promoter expression[J].Proc Natl Acad Sci U S A.2002,99(6): 3663-3667.
    [234]Wang J, Rao S, Chu J, et al.A protein interaction network for pluripotency of embryonic stem cells[J].Nature.2006,444(7117): 364-368.
    [235]Masui S, Nakatake Y, Toyooka Y, et al.Pluripotency governed by Sox2 via regulation of Oct3/4 expression in mouse embryonic stem cells[J].Nat Cell Biol.2007,9(6): 625-635.
    [236]Pesce M, Wang X, Wolgemuth D J, et al.Differential expression of the Oct-4 transcription factor during mouse germ cell differentiation[J].Mech Dev.1998, 71(1-2): 89-98.
    [237]Joshi S, Davies H, Sims L P, et al.Ovarian gene expression in the absence of FIGLA, an oocyte-specific transcription factor[J], BMC Dev Biol.2007, 7: 67.
    [238]Pesce M, Gross M K, Scholer H R.In line with our ancestors: Oct-4 and the mammalian germ[J].Bioessays.1998,20(9): 722-732.
    [239]Western P, Maldonado-saldivia J, Van B J, et al.Analysis of Esgl expression in pluripotent cells and the germline reveals similarities with Oct4 and Sox2 and differences between human pluripotent cell lines[J].Stem Cells.2005,23(10): 1436-1442.
    [240]Esposito M S, Esposito R E.The genetic control of sporulation in Saccharomyces.I.The isolation of temperature-sensitive sporulation-deficient mutants[J].Genetics.1969, 61(1): 79-89.
    [241]Klapholz S, Waddell C S, Esposito R E.The role of the SPOll gene in meiotic recombination in yeast[J].Genetics.1985, 110(2): 187-216.
    [242]Keeney S, Giroux C N, Kleckner N.Meiosis-specific DNA double-strand breaks are catalyzed by Spoil, a member of a widely conserved protein family[J].Cell.1997, 88(3): 375-384.
    [243]Romanienko P J, Camerini-otcro R D.Cloning, characterization, and localization of mouse and human SP011.[J].Genomics.1999,61(2): 156-169.
    [244]Shannon M, Richardson L, Christian A, et al.Differential gene expression of mammalian SPOl 1/TOP6A homologs during meiosis.[J].FEBS Lett.1999,462(3): 329-334.
    [245]Tsubouchi T, Roeder G S.A synaptonemal complex protein promotes homology-independent centromere coupling.[J].Science.2005, 308(5723): 870-873.
    [246]Neale M J, Pan J, Keeney S.Endonucleolytic processing of covalent protein-linked DNA double-strand breaks.[J].Nature.2005,436(7053): 1053-1057.
    [247]Guillon H, Baudat F, Grey C, et al.Crossover and noncrossover pathways in mouse meiosis.[J].Mol Cell.2005,20(4): 563-573.
    [248]Romanienko P J, Camerini-otero R D.The mouse Spol 1 gene is required for meiotic chromosome synapsis.[J].Mol Cell.2000,6(5): 975-987.
    [249J Baudat F, Manova K, Yuen J P, et al.Chromosome synapsis defects and sexually dimorphic meiotic progression in mice lacking Spol 1.[J].Mol Cell.2000, 6(5): 989-998.
    [250]Miura C, Higashino T, Miura T.A progestin and an estrogen regulate early stages of oogenesis in fish[J].Biol Reprod.2007, 77(5): 822-828.
    [251]Ozaki Y, Miura C, Miura T.Molecular cloning and gene expression of Spol 1 during spermatogenesis in the Japanese eel, Anguilla japonica[J].COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY.2006, 143(3): 309-314.
    [252]Ozaki Y, Miura C, Miura T.Molecular cloning and gene expression of Spoil during spermatogenesis in the Japanese eel, Anguilla japonica[J].COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY.2006, 143(3): 309-314.

© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号

地址:北京市海淀区学院路29号 邮编:100083

电话:办公室:(+86 10)66554848;文献借阅、咨询服务、科技查新:66554700