甲基睾酮对雌核发育翘嘴鲌幼鱼性别分化基因表达的影响
|
摘要
为探讨不同质量分数甲基睾酮(Methyltestosterone,MT)对翘嘴鲌幼鱼性别分化的影响,对出膜后4d的雌核发育翘嘴鲌幼鱼进行6和60mg/kg MT拌喂暴露试验。测量幼鱼体长、全长和体质量,组织学方法观察其性腺发育,采用qRT-PCR方法分析其性别决定相关基因(amh、sox9b、dax1、cyp19a1a、foxl2、dmrt1)的表达情况,ELISA方法检测其性激素水平。结果显示,6mg/kg MT处理后,幼鱼全长、体长和体质量均显著增加,在处理后期,60mg/kg MT对体质量也有显著促进作用。组织切片显示,4~60日龄幼鱼均未出现性腺组织。在46~53d的幼鱼发育过程中,除dmrt1外,其他基因表达量均显著上调。MT处理的42~56d,amh、dax1、cyp19a1a和dmrt1的表达均受到抑制,foxl2和sox9b的表达先被抑制后被促进;6mg/kg MT显著升高幼鱼雌二醇(E2)和11-酮基睾酮(11-KT)的水平,60mg/kg MT降低E2水平。在整个暴露期间,睾酮(T)水平与MT质量分数和暴露时间呈反比。结果表明,60日龄雌核发育的翘嘴鲌幼鱼性腺尚未发育成型,46~53日龄是雌核发育翘嘴鲌卵巢发育的关键时期;6和60mg/kg MT对翘嘴鲌幼鱼的生长均有诱导作用,对性腺发育有扰乱作用。
The aim of this study was to investigate the effect of methyltestosterone(MT)on sex differentiation of Culter albumus juveniles,and to provide theoretical guidance for artificial induction of pseudo males.In the present study,the larvae after hatching for 4 days were fed with commercial feed containing 0,6 mg/kg and 60 mg/kg MT for 8 weeks and samples were collected per week.The body length,total length and body mass were measured.Paraffin section was performed to investigate gonad development.The mRNA expressions of sex differentiation related genes including amh,sox9 b,dax1,cyp19 a1 a,foxl2 and dmrt1 were analyzed by qRT-PCR.Sex hormones were detected using ELISA method.The results showed that 6 mg/kg MT significantly increased body length,total length and body mass of C.albumusjuveniles,and 60 mg/kg MT also significantly increased the body mass at the end of treatment.The histology section results showed that the gonad tissues did not appear in both control and MT treated groups until 60 days after hatching(DAH).During the development of the juveniles,the mRNA expressions of the sex differentiation related genes were significantly up-regulated from 46 to 53 DAH except for dmrt1 gene.In later period of MT exposure(42-56 DAH),the mRNA expression of amh,dax1,cyp19 a1 aand dmrt1 was significantly inhibited,however the mRNA levels of foxl2 and sox9 b were decreased at 42 DAH first and subsequently increased at 49 DAH and56 DAH following MT exposure.In the later treatment period(42-56 DAH),estradiol level was increased in MT treatmentat low concentration(6 mg/kg)and decreased in MT treatment athigh concentration(60 mg/kg).During the whole exposure duration,the testosterone level was decreased in both time and dose dependent manner,while 11-ketotestosterone level was increased in MT treatment at low concentration(6 mg/kg).Therefore,these results suggested that the gonads had not developed in60 DAH,the period from 46 to 53 DAH was the critical stage for sex differentiation of gynogenetic C.albumus juveniles.In addition,6 mg/kg and 60 mg/kg MT could accelerate growth and disturb sex differentiation in the juveniles.
The aim of this study was to investigate the effect of methyltestosterone(MT)on sex differentiation of Culter albumus juveniles,and to provide theoretical guidance for artificial induction of pseudo males.In the present study,the larvae after hatching for 4 days were fed with commercial feed containing 0,6 mg/kg and 60 mg/kg MT for 8 weeks and samples were collected per week.The body length,total length and body mass were measured.Paraffin section was performed to investigate gonad development.The mRNA expressions of sex differentiation related genes including amh,sox9 b,dax1,cyp19 a1 a,foxl2 and dmrt1 were analyzed by qRT-PCR.Sex hormones were detected using ELISA method.The results showed that 6 mg/kg MT significantly increased body length,total length and body mass of C.albumusjuveniles,and 60 mg/kg MT also significantly increased the body mass at the end of treatment.The histology section results showed that the gonad tissues did not appear in both control and MT treated groups until 60 days after hatching(DAH).During the development of the juveniles,the mRNA expressions of the sex differentiation related genes were significantly up-regulated from 46 to 53 DAH except for dmrt1 gene.In later period of MT exposure(42-56 DAH),the mRNA expression of amh,dax1,cyp19 a1 aand dmrt1 was significantly inhibited,however the mRNA levels of foxl2 and sox9 b were decreased at 42 DAH first and subsequently increased at 49 DAH and56 DAH following MT exposure.In the later treatment period(42-56 DAH),estradiol level was increased in MT treatmentat low concentration(6 mg/kg)and decreased in MT treatment athigh concentration(60 mg/kg).During the whole exposure duration,the testosterone level was decreased in both time and dose dependent manner,while 11-ketotestosterone level was increased in MT treatment at low concentration(6 mg/kg).Therefore,these results suggested that the gonads had not developed in60 DAH,the period from 46 to 53 DAH was the critical stage for sex differentiation of gynogenetic C.albumus juveniles.In addition,6 mg/kg and 60 mg/kg MT could accelerate growth and disturb sex differentiation in the juveniles.
引文
[1]王伟.翘嘴鲌(Culter albunus)群体遗传多样性及鲌亚科鱼类系统发生的研究[D].上海:华东师范大学,2007.WANG W.Study on population genetie diversity of(Culter albunus)and phylogeny of Cultrinae[D].Shanghai:East China Normal University,2007.
[2]李停,贾永义,刘士力,等.二次正交旋转组合设计优化人工诱导翘嘴鲌雌核发育[J].中国水产科学,2016,23(1):77-89.LI T,JIA Y Y,LIU SH L,et al.Optimizing conditions to induce artificial gynogenesis in Culter alburnus using a quadratic orthogonal rotation design[J].Journal of Fishery Sciences of China,2016,23(1):77-89.
[3]徐竹青.牙鲆全雌化研究的新进展[J].中国水产,2005(8):80-80.XU ZH Q.New process of feminization of Bastard halibut[J].China Fisheries,2005(8):80-80.
[4]郑尧,王在照,陈家长.调控鱼类性腺分化基因的研究进展[J].水生生物学报,2015,39(4):798-810.ZHENG Y,WANG Z ZH,CHEN J ZH.Progressess on the study of sex difference genes in fish[J].Acta Hydrobiologica Sinica,2015,39(4):798-810.
[5]JRGENSEN A,MORTHORST J E,ANDERSEN O,et al.Expression profiles for six zebrafish genes during gonadal sex differentiation[J].Reproductive Biology and Endocrinology,2008,6(5):181-190.
[6]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.WANG D SH,WU T L.Fish sex determination and its mechanisms:a review[J].Journal of Southwest China Normal University(Natural Science Edition),2000,25(3):296-304.
[7]GIUILI G,SHEN W H,INGRAHAM H A.The nuclear receptor SF-1 mediates sexually dimorphic expression of Müullerian inhibiting substance,in vivo[J].Development,1997,124(9):1799-1807.
[8]LEE M M,DONAHOE P K.Müllerian inhibiting substance:agonadal hormone with multiple functions[J].Endocrine Reviews,1993,14(2):152-164.
[9]MNSTERBERG A,LOVELLBADGE R.Expression of the mouse anti-mullerian hormone gene suggests a role in both male and female sexual differentiation[J].Development,1991,113(2):613-624.
[10]LANE,ANDREW H,LEE,et al.Müllerian inhibiting substance:a nontraditional marker of gonadal function[J].Current Opinion in Endocrinology Diabetes and Obesity,2001,8(6):296-300.
[11]MIURA T,MIURA C,KONDA Y,et al.Spermatogenesispreventing substance in Japanese eel[J].Development,2002,129(11):2689-2697.
[12]文爱韵,尤锋,徐永立,等.鱼类性别决定与分化相关基因研究进展[J].海洋科学,2008,32(1):74-80.WEN A Y,YOU F,XU Y L,et al.Research progress on the study of sex determination and differentiation relateds to gene of fish[J].Marine Sciences,2008,32(1):74-80.
[13]DEVLIN R H,NAGAHAMA Y,DEVLIN R H,et al.Sex determination and sex differentiation in fish:an overview of genetic,physiological,and environmental influences[J].Aquaculture,2002,208(3/4):191-364.
[14]ZHANG H,THOMSEN J S,JOHANSSON L,et al.DAX-1functions as an LXXLL-containing corepressor for activated estrogen receptors[J].Journal of Biological Chemistry,2000,275(51):39855-39859.
[15]HOLTER E,KOTAJA N,MAKELA S,et al.Inhibition of androgen receptor(AR)function by the reproductive orphan nuclear receptor DAX-1[J].Molecular Endocrinology,2002,16(3):515-528.
[16]WANG D S,KOBAYASHI T,SENTHILKUMARAN B,et al.Molecular cloning of DAX1 and SHP cDNAs and their expression patterns in the Nile tilapia,Oreochromis niloticus[J].Biochemical and Biophysical Research Communications,2002,297(3):632-640.
[17]BARON D,HOULGATTE R,FOSTIER A,et al.Largescale temporal gene expression profiling during gonadal differentiation and early gametogenesis in rainbow trout[J].Biology of Reproduction,2005,73(5):959-966.
[18]CARLSSON P,MAHLAPUU M F.Transcription factors:key players in development and metabolism[J].Developmental Biology,2002,250(1):1-23.
[19]SRIDEVI P,CHAITANYA R K,DUTTA-GUPTA A,et al.FTZ-F1 and FOXL2 up-regulate catfish brain aromatase gene transcription by specific binding to the promoter motifs[J].Biochimicaet Biophysica Acta-Biomembranes,2012,1819(1):57-66.
[20]YOSHIURA Y,SENTHILKUMARAN B,WATANABEM,et al.Synergistic expression of Ad4BP/SF-1and cytochrome P-450aromatase(ovarian type)in the ovary of Nile tilapia,Oreochromis niloticus,during vitellogenesis suggests transcriptional interaction[J].Biology of Reproduction,2003,68(5):1545-1553.
[21]张悦,鲁晓萱,单祥年.性别决定基因的研究进展[J].遗传,2000,22(5):328-330.ZHANG Y,LU X X,SHAN X N.Progress in research on sex determinating genes[J].Hereditas,2000,22(5):328-330.
[22]CHIANG F L,PAI C I,WYATT M,et al.Twosox9 genes on duplicated zebrafish chromosomes:expression of similar transcription activators in distinct sites[J].Developmental Biology,2001,231(1):149-163.
[23]陈芸,王艺磊,田佳,等.斑马鱼性别决定相关基因的表达分析[J].水生态学杂志,2010,3(5):10-16.CHEN Y,WANG Y L,TIAN J,et al.Expression analysis of genes which related to sex determination of zebrafish[J].Reservoir Fisheries,2010,3(5):10-16.
[24]VANDESOMPELE J,PRETER K D,PATTYN F,et al.Accurate normalization of real-time quantitative RT-PCRdata by geometric averaging of multiple internal control genes[J].Genome Biology,2002,3(7):0034.1-0034.11.
[25]HIGGS D A,FAGERLUND U H,MCBRIDE J R,et al.Influence of combinations of bovine growth hormone,17alpha-methyltestosterone,and L-thyroxine on growth of yearling coho salmon(Oncorhynchus kisutch)[J].Canadian Journal of Zoology,1977,55(6):1048-56.
[26]马尚助,于凌杰,刘明波,等.应用17α-甲基睾丸酮促进虹鳟稚鱼生长的试验[J].淡水渔业,1990(4):6-7.MA SH ZH,YU L J,LIU M B,et al.The growth experiment of 17 alpha methyl testosterone Oncorhynchus mykiss juvenile[J].Freshwater Fisheries,1990(4):6-7.
[27]任维美.投喂含雄性激素饲料可有效提高鲤鱼产量[J].国外水产,1989(3):45-45.REN W M.Effectively improve the production of feeding containing testosterone feed carp[J].Foreign Fisheries Periodicals,1989(3):45-45.
[28]王晶,王冰,李纪同,等.斑马鱼性腺发育的组织学观察[J].基因组学与应用生物学,2011,30(2):168-174.WANG J,WANG B,LI J T,et al.Histological observation of zebrafish gonad development[J].Genomics and Applied Biology,2011,30(2):168-174.
[29]彭丽娜,卢建国,栾培贤,等.黄颡鱼(Pelteobagrus fulvidraco)性腺分化的组织学观察[J].水产学杂志,2014,27(3):48-51.PENG L N,LU J G,LUAN P X,et al.Observation on gonadal differentiation in Yellow Catfish Pelteobagrus fulvidraco[J].Chinese Journal of Fisheries,2014,27(3):48-51.
[30]耿智.黄姑鱼早期生长和性腺发育研究[D].浙江舟山:浙江海洋学院,2012.GENG ZH.Research on the early growth and gonad development of the spotted maigre Nibea albiflora[D].Zhou shan Zhejiang:Zhejiang Ocean University,2012.
[31]马学坤,柳学周,温海深,等.半滑舌鳎性腺分化的组织学观察[J].渔业科学进展,2006,27(2):55-61.MA X K,LIU X ZH,WEN H SH,et al.Histological observation on gonadal sex differentiation in Cynoglossus semilaevis Gunther[J].Marine Fisheries Research,2006,27(2):55-61.
[32]刘姗姗.半滑舌鳎AMH基因的克隆及表达分析[D].上海:上海海洋大学,2012.LIU SH SH.Molecular cloning and expression analysis of AMH gene from Cynoglossus semilaevis Gunther[D].Shanghai:Shanghai Ocean University,2012.
[33]蒋香菊,吴阳升,张利,等.羔羊卵巢AMH的免疫组织化学分析[J].西北农业学报,2014,23(7):19-24.JIANG X J,WU Y SH,ZHANG L,et al.Immunocy tochemical study of anti-Miillerianhormone in ovarian follicles from lambs[J].Acta Agriculturae Boreali-occidentalis Sinica,2014,23(7):19-24.
[34]邓思平,陈松林.半滑舌鳎Dmrt1α基因的cDNA克隆及其表达[J].中国水产科学,2008,15(4):577-584.DENG S P,CHEN S L.Molecular cloning and expression of Dmrt1αgene fromCynoglossus semilaevis Gunther[J].Journal of Fishery Sciences of China,2008,15(4):577-584.
[35]王晶晶,刘小林,秦芳,等.稀有鮈鲫芳香化酶基因的表达及内分泌干扰物(EDCs)暴露对其表达的影响[J].西北农业学报,2011,20(1):35-39.WANG J J,LIU X L,QIN F,et al.Expression of cytochrome P450aromatase genes and effects of EDCs exposure on their expression in rare minnow(Gobiocypris rarus)[J].Acta Agriculturae Boreali-occidentalis Sinica,2011,20(1):35-39.
[36]WANG D S,KOBAYASHI T,ZHOU L Y,et al.Foxl2 upregulates aromatase gene transcription in a female-specific manner by binding to the promoter as well as interacting with ad4binding protein/steroidogenic factor 1[J].Molecular Endocrinology,2007,21(3):712-725.
[37]MARINA S G,MALLE P,PAILHOUX E,et al.Isolation of chicken homolog of the FOXL2,gene and comparison of its expression patterns with those of aromatase during ovarian development[J].Developmental Dynamics,2004,231(4):859-870.
[38]邬国民,练慧英.应用甲基睾丸素诱导莫桑非洲鲫雄性化的研究[J].遗传,1979,1(6):36-39.WU G M,LIAN H Y.Research on the application of methyl testosterone inducing mozambique’s African carp male-like[J].Hereditas,1979,1(6):36-39.
[39]张甫英,胡炜,汪亚平,等.应用雄性激素诱导异育银鲫性转化的研究[J].遗传,2000,22(1):25-27.ZHANG F Y,HU W,WANG Y P,et al.Methyltestosteroneinduction of sex reversal in allogynogenetic crucian carp[J].Hereditas,2000,22(1):25-27.
[2]李停,贾永义,刘士力,等.二次正交旋转组合设计优化人工诱导翘嘴鲌雌核发育[J].中国水产科学,2016,23(1):77-89.LI T,JIA Y Y,LIU SH L,et al.Optimizing conditions to induce artificial gynogenesis in Culter alburnus using a quadratic orthogonal rotation design[J].Journal of Fishery Sciences of China,2016,23(1):77-89.
[3]徐竹青.牙鲆全雌化研究的新进展[J].中国水产,2005(8):80-80.XU ZH Q.New process of feminization of Bastard halibut[J].China Fisheries,2005(8):80-80.
[4]郑尧,王在照,陈家长.调控鱼类性腺分化基因的研究进展[J].水生生物学报,2015,39(4):798-810.ZHENG Y,WANG Z ZH,CHEN J ZH.Progressess on the study of sex difference genes in fish[J].Acta Hydrobiologica Sinica,2015,39(4):798-810.
[5]JRGENSEN A,MORTHORST J E,ANDERSEN O,et al.Expression profiles for six zebrafish genes during gonadal sex differentiation[J].Reproductive Biology and Endocrinology,2008,6(5):181-190.
[6]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.WANG D SH,WU T L.Fish sex determination and its mechanisms:a review[J].Journal of Southwest China Normal University(Natural Science Edition),2000,25(3):296-304.
[7]GIUILI G,SHEN W H,INGRAHAM H A.The nuclear receptor SF-1 mediates sexually dimorphic expression of Müullerian inhibiting substance,in vivo[J].Development,1997,124(9):1799-1807.
[8]LEE M M,DONAHOE P K.Müllerian inhibiting substance:agonadal hormone with multiple functions[J].Endocrine Reviews,1993,14(2):152-164.
[9]MNSTERBERG A,LOVELLBADGE R.Expression of the mouse anti-mullerian hormone gene suggests a role in both male and female sexual differentiation[J].Development,1991,113(2):613-624.
[10]LANE,ANDREW H,LEE,et al.Müllerian inhibiting substance:a nontraditional marker of gonadal function[J].Current Opinion in Endocrinology Diabetes and Obesity,2001,8(6):296-300.
[11]MIURA T,MIURA C,KONDA Y,et al.Spermatogenesispreventing substance in Japanese eel[J].Development,2002,129(11):2689-2697.
[12]文爱韵,尤锋,徐永立,等.鱼类性别决定与分化相关基因研究进展[J].海洋科学,2008,32(1):74-80.WEN A Y,YOU F,XU Y L,et al.Research progress on the study of sex determination and differentiation relateds to gene of fish[J].Marine Sciences,2008,32(1):74-80.
[13]DEVLIN R H,NAGAHAMA Y,DEVLIN R H,et al.Sex determination and sex differentiation in fish:an overview of genetic,physiological,and environmental influences[J].Aquaculture,2002,208(3/4):191-364.
[14]ZHANG H,THOMSEN J S,JOHANSSON L,et al.DAX-1functions as an LXXLL-containing corepressor for activated estrogen receptors[J].Journal of Biological Chemistry,2000,275(51):39855-39859.
[15]HOLTER E,KOTAJA N,MAKELA S,et al.Inhibition of androgen receptor(AR)function by the reproductive orphan nuclear receptor DAX-1[J].Molecular Endocrinology,2002,16(3):515-528.
[16]WANG D S,KOBAYASHI T,SENTHILKUMARAN B,et al.Molecular cloning of DAX1 and SHP cDNAs and their expression patterns in the Nile tilapia,Oreochromis niloticus[J].Biochemical and Biophysical Research Communications,2002,297(3):632-640.
[17]BARON D,HOULGATTE R,FOSTIER A,et al.Largescale temporal gene expression profiling during gonadal differentiation and early gametogenesis in rainbow trout[J].Biology of Reproduction,2005,73(5):959-966.
[18]CARLSSON P,MAHLAPUU M F.Transcription factors:key players in development and metabolism[J].Developmental Biology,2002,250(1):1-23.
[19]SRIDEVI P,CHAITANYA R K,DUTTA-GUPTA A,et al.FTZ-F1 and FOXL2 up-regulate catfish brain aromatase gene transcription by specific binding to the promoter motifs[J].Biochimicaet Biophysica Acta-Biomembranes,2012,1819(1):57-66.
[20]YOSHIURA Y,SENTHILKUMARAN B,WATANABEM,et al.Synergistic expression of Ad4BP/SF-1and cytochrome P-450aromatase(ovarian type)in the ovary of Nile tilapia,Oreochromis niloticus,during vitellogenesis suggests transcriptional interaction[J].Biology of Reproduction,2003,68(5):1545-1553.
[21]张悦,鲁晓萱,单祥年.性别决定基因的研究进展[J].遗传,2000,22(5):328-330.ZHANG Y,LU X X,SHAN X N.Progress in research on sex determinating genes[J].Hereditas,2000,22(5):328-330.
[22]CHIANG F L,PAI C I,WYATT M,et al.Twosox9 genes on duplicated zebrafish chromosomes:expression of similar transcription activators in distinct sites[J].Developmental Biology,2001,231(1):149-163.
[23]陈芸,王艺磊,田佳,等.斑马鱼性别决定相关基因的表达分析[J].水生态学杂志,2010,3(5):10-16.CHEN Y,WANG Y L,TIAN J,et al.Expression analysis of genes which related to sex determination of zebrafish[J].Reservoir Fisheries,2010,3(5):10-16.
[24]VANDESOMPELE J,PRETER K D,PATTYN F,et al.Accurate normalization of real-time quantitative RT-PCRdata by geometric averaging of multiple internal control genes[J].Genome Biology,2002,3(7):0034.1-0034.11.
[25]HIGGS D A,FAGERLUND U H,MCBRIDE J R,et al.Influence of combinations of bovine growth hormone,17alpha-methyltestosterone,and L-thyroxine on growth of yearling coho salmon(Oncorhynchus kisutch)[J].Canadian Journal of Zoology,1977,55(6):1048-56.
[26]马尚助,于凌杰,刘明波,等.应用17α-甲基睾丸酮促进虹鳟稚鱼生长的试验[J].淡水渔业,1990(4):6-7.MA SH ZH,YU L J,LIU M B,et al.The growth experiment of 17 alpha methyl testosterone Oncorhynchus mykiss juvenile[J].Freshwater Fisheries,1990(4):6-7.
[27]任维美.投喂含雄性激素饲料可有效提高鲤鱼产量[J].国外水产,1989(3):45-45.REN W M.Effectively improve the production of feeding containing testosterone feed carp[J].Foreign Fisheries Periodicals,1989(3):45-45.
[28]王晶,王冰,李纪同,等.斑马鱼性腺发育的组织学观察[J].基因组学与应用生物学,2011,30(2):168-174.WANG J,WANG B,LI J T,et al.Histological observation of zebrafish gonad development[J].Genomics and Applied Biology,2011,30(2):168-174.
[29]彭丽娜,卢建国,栾培贤,等.黄颡鱼(Pelteobagrus fulvidraco)性腺分化的组织学观察[J].水产学杂志,2014,27(3):48-51.PENG L N,LU J G,LUAN P X,et al.Observation on gonadal differentiation in Yellow Catfish Pelteobagrus fulvidraco[J].Chinese Journal of Fisheries,2014,27(3):48-51.
[30]耿智.黄姑鱼早期生长和性腺发育研究[D].浙江舟山:浙江海洋学院,2012.GENG ZH.Research on the early growth and gonad development of the spotted maigre Nibea albiflora[D].Zhou shan Zhejiang:Zhejiang Ocean University,2012.
[31]马学坤,柳学周,温海深,等.半滑舌鳎性腺分化的组织学观察[J].渔业科学进展,2006,27(2):55-61.MA X K,LIU X ZH,WEN H SH,et al.Histological observation on gonadal sex differentiation in Cynoglossus semilaevis Gunther[J].Marine Fisheries Research,2006,27(2):55-61.
[32]刘姗姗.半滑舌鳎AMH基因的克隆及表达分析[D].上海:上海海洋大学,2012.LIU SH SH.Molecular cloning and expression analysis of AMH gene from Cynoglossus semilaevis Gunther[D].Shanghai:Shanghai Ocean University,2012.
[33]蒋香菊,吴阳升,张利,等.羔羊卵巢AMH的免疫组织化学分析[J].西北农业学报,2014,23(7):19-24.JIANG X J,WU Y SH,ZHANG L,et al.Immunocy tochemical study of anti-Miillerianhormone in ovarian follicles from lambs[J].Acta Agriculturae Boreali-occidentalis Sinica,2014,23(7):19-24.
[34]邓思平,陈松林.半滑舌鳎Dmrt1α基因的cDNA克隆及其表达[J].中国水产科学,2008,15(4):577-584.DENG S P,CHEN S L.Molecular cloning and expression of Dmrt1αgene fromCynoglossus semilaevis Gunther[J].Journal of Fishery Sciences of China,2008,15(4):577-584.
[35]王晶晶,刘小林,秦芳,等.稀有鮈鲫芳香化酶基因的表达及内分泌干扰物(EDCs)暴露对其表达的影响[J].西北农业学报,2011,20(1):35-39.WANG J J,LIU X L,QIN F,et al.Expression of cytochrome P450aromatase genes and effects of EDCs exposure on their expression in rare minnow(Gobiocypris rarus)[J].Acta Agriculturae Boreali-occidentalis Sinica,2011,20(1):35-39.
[36]WANG D S,KOBAYASHI T,ZHOU L Y,et al.Foxl2 upregulates aromatase gene transcription in a female-specific manner by binding to the promoter as well as interacting with ad4binding protein/steroidogenic factor 1[J].Molecular Endocrinology,2007,21(3):712-725.
[37]MARINA S G,MALLE P,PAILHOUX E,et al.Isolation of chicken homolog of the FOXL2,gene and comparison of its expression patterns with those of aromatase during ovarian development[J].Developmental Dynamics,2004,231(4):859-870.
[38]邬国民,练慧英.应用甲基睾丸素诱导莫桑非洲鲫雄性化的研究[J].遗传,1979,1(6):36-39.WU G M,LIAN H Y.Research on the application of methyl testosterone inducing mozambique’s African carp male-like[J].Hereditas,1979,1(6):36-39.
[39]张甫英,胡炜,汪亚平,等.应用雄性激素诱导异育银鲫性转化的研究[J].遗传,2000,22(1):25-27.ZHANG F Y,HU W,WANG Y P,et al.Methyltestosteroneinduction of sex reversal in allogynogenetic crucian carp[J].Hereditas,2000,22(1):25-27.