泥鳅精子和卵子结构及受精过程的细胞学观察
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摘要
研究旨在探讨泥鳅精子受精时限及为其人工繁殖提供基础资料。采用光镜、电镜技术对泥鳅(Misgurnus anguillicaudatus)精子、卵子和不同时间段的受精卵进行了观察。结果显示:泥鳅精子头部无顶体,主要为核占据,核凹窝较浅,中段具不对称的袖套,尾部轴丝为"9+2"结构,无侧鳍。卵子动物极卵膜仅有一受精孔,受精孔为深凹陷、短孔道型。在20—21℃水温条件下,授精后3s,精子开始穿过精孔管或已经进入卵子;授精后5—8s,形成精子星光;授精后70s,卵子处于第二次减数分裂后期;授精后6—8min,第二极体形成,等待排出;授精后20—25min,雌雄原核融合;授精后25—30min,受精卵进入第一次有丝分裂中期;授精后40—45min,第一次有丝分裂结束,二细胞形成。研究表明:成熟卵子精孔管内口径(2.286±0.364)μm,而精子头部直径与其接近,单精受精;入水后150s内可受精。
This study investigated fertilization of spermatozoa of Misgurnus. anguillicaudatus at different time periods using optical microscope and electron microscope technology methods. The results showed that no acrosome was observed in the head of the sperm, which was mainly occupied by the nuclear. The implantation fossa of the sperms was low, and the mid-piece contained asymmetric sleeve. The flagella without lateral fins were organized as a "9+2" microtubular structure by axonemes. There was only one micropyle in shell of egg's animal pole, and the micropyle belongs to the type of concave and short channel. Under the water temperature of 20—21, sperm passed micropylar tube and entered ovum at 3s after insemination; sperm star light was observed at 5—8s; the second meiosis of the ovum occurred at 70 s after insemination; the second polocyte appeared at 6—8min after insemination. Male and female prnucleus were fused at 20—25min after insemination. Zygote was in metaphase of first mitosis at 25—30min after insemination. The first mitosis finished at 40—45min after insemination, and two cells then formed. The inner diameter of micropylar tube of mature egg was(2.286±0.364) μm, similar like the diameter of sperm head, and the monospermy of M. anguillicaudatus. Eggs of M. anguillicaudatus were fertilizable in 150 s after entering to water.
This study investigated fertilization of spermatozoa of Misgurnus. anguillicaudatus at different time periods using optical microscope and electron microscope technology methods. The results showed that no acrosome was observed in the head of the sperm, which was mainly occupied by the nuclear. The implantation fossa of the sperms was low, and the mid-piece contained asymmetric sleeve. The flagella without lateral fins were organized as a "9+2" microtubular structure by axonemes. There was only one micropyle in shell of egg's animal pole, and the micropyle belongs to the type of concave and short channel. Under the water temperature of 20—21, sperm passed micropylar tube and entered ovum at 3s after insemination; sperm star light was observed at 5—8s; the second meiosis of the ovum occurred at 70 s after insemination; the second polocyte appeared at 6—8min after insemination. Male and female prnucleus were fused at 20—25min after insemination. Zygote was in metaphase of first mitosis at 25—30min after insemination. The first mitosis finished at 40—45min after insemination, and two cells then formed. The inner diameter of micropylar tube of mature egg was(2.286±0.364) μm, similar like the diameter of sperm head, and the monospermy of M. anguillicaudatus. Eggs of M. anguillicaudatus were fertilizable in 150 s after entering to water.
引文
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[7]Gao J,Koshio S,Wang W,et al.Effects of dietary phospholipid levels on growth performance,fatty acid composition and antioxidant responses of Dojo loach Misgurnus anguillicaudatus Larvae[J].Aquaculture,2014,426:304-309
[8]Zhou X,Yu Y,Li Y,et al.Comparative analysis of mitochondrial genomes in distinct nuclear ploidy loach Misgurnus anguillicaudatus and its implications for polyploidy evolution[J].Plos One,2014,9(3):e92033
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[11]Kang X J,Liang C G,Guo M S,et al.Ultrastructure of spermatogenesis and spermiogenesis in the half-smooth tongue sole Cynoglossus semilaevis[J].Acta Zoologica Sinica,2008,54(2):356-365[康现江,梁春光,郭明申,等.半滑舌鳎精子发生和精子形成的超微结构.动物学报,2008,54(2):356-365]
[12]You Y L,Lin D J.A study on the fine structure of the spermatozoon of teleost Cyprinus carpio(Linnaeus)[J].Zoological Research,1996,17(4):377-383[尤永隆,林丹军.鲤鱼精子超微结构的研究.动物学研究,1996,17(4):377-383]
[13]Hu J,Zhang Y,Fu C,et al.Ultrastructure of rosy barb Puntius conchonius spermatozoon[J].Acta Zoologica Sinica,2005,51(5):892-897
[14]Li F,Wan Q,Huang X M.Study on the structure of Paramisgurnus dabryanus spermatozoa[J].Journal of Biology,2011,28(4):26-30[李飞,万全,黄鲜明.大鳞副泥鳅精子结构研究.生物学杂志,2011,28(4):26-30]
[15]Zhang Y G,Luo Q S,Zhong M C.Studies on the structure of testis and spermatozoon in Leiocassis longirostris[J].Acta Hydrobiologica Sinica,1993,17(3):246-251[张耀光,罗泉笙,钟明超.长吻鮠精巢及精子结构的研究.水生生物学报,1993,17(3):246-251]
[16]You Y L,Lin D J.The ultrastructure of spermatozoon of the teleost Pseudobagrus fulvidraco[J].Acta Biologiae Experimentalis Sinica,1996,29(3):235-245[尤永隆,林丹军.黄颡鱼(Pseudobagrus fulvidraco)精子的超微结构.实验生物学报,1996,29(3):235-245]
[17]Gwo J C,Kuo M C,Chiu J Y,et al.Ultrastructure of Pagrus major and Rhabdosargus sarba spermatozoa(Perciformes:Sparidae:Sparinae)[J].Tissue and Cell,2004,36(2):141-147
[18]Gwo J C,Yang W T,Kuo M C,et al.Spermatozoal ultrastructures of two marine perciform teleost fishes,the goatfish,Paraupeneus spilurus(Mullidae)and the rabbitfish,Siganus fuscescens(Siganidae)from Taiwan[J].Tissue and Cell,2004,36(1):63-69
[19]Gwo J C,Chiu J Y,Lin C Y,et al.Spermatozoal ultrastructure of four Sparidae fishes:Acanthopagrus berda,Acanthopagrus australis,Lagodon rhomboids and Archosargus probatocephus[J].Tissue and Cell,2005,37(2):109-115
[20]Linhart O,Kudo S.Surface ultrastructure of paddlefish eggs before and after fertilization[J].Journal of Fish Biology,1997,51(3):573-582
[21]Xu Y,Xiong Q M.Cytological study of the fertilization of Acipenser sinensis Gray[J].Acta Zoologica Sinica,1990,36(3):275-278[许雁,熊全沬.中华鲟受精细胞学研究.动物学报,1990,36(3):275-279]
[22]Sun J J,Guo Y G,Li G F,et al.An observation on the early fertilization of Squaliobarbus curriculus by scanning electron microscope[J].Journal of Fishery Sciences of China,2006,13(5):740-744[孙际佳,郭云贵,李桂峰,等.赤眼鳟精子入卵的扫描电镜观察.中国水产科学,2006,13(5):740-744]
[23]Kudo S,Linhart O,Billard R.Ultrastructural studies of sperm penetration in the egg of the European catfish,Silurus glanis[J].Aquatic Living Resources,1994,7(2):93-98
[24]Fu C J,Liu X T,Shan J,et al.Cytological study on maturity of egg in Ctenopharyngodon idllus[J].Freshwater Fisheries,1981,(5):5-8[傅朝君,刘宪亭,单健,等.草鱼卵球成熟细胞学的研究.淡水渔业,1981,(5):5-8]
[25]Huang Y S.Cytological study on fertilization of egg in Tilapia nilotica[J].Acta Zoologica Sinica,1993,39(1):19-22[黄永松.尼罗罗非鱼卵母细胞受精细胞学研究.动物学报,1993,39(1):19-22]
[26]Wang R X,Zhang Y R,Fu C S,et al.The observation on the initial fertilization of freshwater bream by electronic scanning microscope[J].Journal of Fisheries of China,1982,6(4):313-320[王瑞霞,张毓人,傅仓生,等.鲂鱼受精早期精子入卵的扫描电子显微镜观察.水产学报,1982,6(4):313-320]
[27]Li P,Liu X L,Li Y,et al.Cytological study on fertilization process of Myxocyprinus asiaticus[J].Journal of Fishery Sciences of China,2013,20(2):250-259[李萍,刘晓蕾,李勇,等.胭脂鱼受精过程的细胞学观察.中国水产科学,2013,20(2):250-259]
[28]Shen Q Z,Wu K Z,Cai Z Y.Dynamical action of sperm penetration into the egg of Misgurnus anguillicaudatus[J].Zoological Research,1990,11(3):179-184[沈其璋,吴坤璋,蔡振岩.泥鳅精子入卵的动力作用.动物学研究,1990,11(3):179-184]
[29]Gan G M.Studies on Fertilization Biology of Hemibarbus labeo[D].Southwest University,2003[甘光明.唇?受精生物学研究.西南大学,2003]
[30]Wang Y M.Studies on the Fertilization Biology of Gobiocypris rarus[D].Southwest University,2011[王永明.稀有逗鲫受精生物学研究.西南大学,2011]
[31]Yao J H,Zhou P F.Scanning electron microscopic observation on the ultrastructure of the chorion and the process of insemination of Paramisgurnus dabryanus[J].Journal of Shanghai Fisheries University,1998,7(1):65-68[姚纪花,周平凡.大鳞副泥鳅卵子壳膜结构与授精过程的扫描电镜观察.上海水产大学学报,1998,7(1):65-68]
[32]Xu R W.An observation on the lifetime of loach M.anguillicaudatus spermatozoa[J].Journal of Zhejiang College of Fisheries(Natural Science Edition),1992,11(1):17[徐如卫.泥鳅Misgurnus anguillicaudatus Cantor精子寿命的初步观察.浙江海洋学院学报(自然科学版),1992,11(1):17]
[33]Zhang T,Liu L,Zhang J M,et al.Ultrastructure of spermatozoa and fertilized eggs of Anguilla japonica and observation on the fertilization process[J].Acta Hydrobiologica Sinica,2010,34(4):769-778[张涛,柳凌,张洁明,等.日本鳗鲡精卵的超微结构以及受精过程观察.水生生物学报,2010,34(4):769-778]
[2]Huang J,Yang S,Qin Z B,et al.Comparative study about flesh contents and nutrient values in brown bullhead,loach and darkbarbel catfish[J].Acta Hydrobiologica Sinica,2010,34(5):990-997[黄钧,杨淞,覃志彪,等.云斑鲖、泥鳅和瓦氏黄颡鱼的含肉率及营养价值比较研究.水生生物学报,2010,34(5):990-997]
[3]Du Z C.Artificial breeding and seedling culture technique for Misgurnus anguillicaudatus[J].Freshwater Fisheries,2003,33(6):57-58[杜忠臣.泥鳅人工繁殖及苗种培育技术.淡水渔业,2003,33(6):57-58]
[4]Wu P,He Z Z,Wei Y H,et al.Analysis of nutrition in young triploid loach Misgurnus anguillicaudatus[J].Journal of Fisheries of China,2007,31(B9):100-103[吴萍,何珠子,魏育红,等.三倍体泥鳅幼鱼的营养分析.水产学报,2007,31(B9):100-103]
[5]Mcmahon B R,Burggren W W.Respiratory physiology of intestinal air breathing in the teleost fish Misgurnus anguillicaudatus[J].Journal of Experimental Biology,1987,133(1):371-393
[6]Tang J X,Tang Y Y,Sun H X,et al.Effects of Cu2+and Pb2+(single factor and joint toxicity)on DNA damage in Misgurnus anguillicaudatus oocytes[J].Acta Hydrobiologica Sinica,2013,37(3):501-506[唐建勋,唐奕扬,孙红祥,等.重金属Cu2+、Pb2+单因子及联合毒性对泥鳅卵细胞DNA的损伤效应.水生生物学报,2013,37(3):501-506]
[7]Gao J,Koshio S,Wang W,et al.Effects of dietary phospholipid levels on growth performance,fatty acid composition and antioxidant responses of Dojo loach Misgurnus anguillicaudatus Larvae[J].Aquaculture,2014,426:304-309
[8]Zhou X,Yu Y,Li Y,et al.Comparative analysis of mitochondrial genomes in distinct nuclear ploidy loach Misgurnus anguillicaudatus and its implications for polyploidy evolution[J].Plos One,2014,9(3):e92033
[9]Chen D Y.Fertilization Biology:Fertilization Mechanism and Reproductive Engineering[M].Beijing:Science Press.2000,2-12[陈大元.受精生物学:受精机制与生殖工程.北京:科学出版社,2000,2-12]
[10]Du C L,Zhao N,Wang Y M,et al.Studies on the ultra/microscopic structures of spermatozoon in Megalobrama pellegrini[J].Journal of Southwest China Normal University(Natural Science Edition),2012,37(2):72-76[杜长雷,赵娜,王永明,等.厚颌鲂精子的显微及超微结构观察.西南师范大学学报(自然科学版),2012,37(2):72-76]
[11]Kang X J,Liang C G,Guo M S,et al.Ultrastructure of spermatogenesis and spermiogenesis in the half-smooth tongue sole Cynoglossus semilaevis[J].Acta Zoologica Sinica,2008,54(2):356-365[康现江,梁春光,郭明申,等.半滑舌鳎精子发生和精子形成的超微结构.动物学报,2008,54(2):356-365]
[12]You Y L,Lin D J.A study on the fine structure of the spermatozoon of teleost Cyprinus carpio(Linnaeus)[J].Zoological Research,1996,17(4):377-383[尤永隆,林丹军.鲤鱼精子超微结构的研究.动物学研究,1996,17(4):377-383]
[13]Hu J,Zhang Y,Fu C,et al.Ultrastructure of rosy barb Puntius conchonius spermatozoon[J].Acta Zoologica Sinica,2005,51(5):892-897
[14]Li F,Wan Q,Huang X M.Study on the structure of Paramisgurnus dabryanus spermatozoa[J].Journal of Biology,2011,28(4):26-30[李飞,万全,黄鲜明.大鳞副泥鳅精子结构研究.生物学杂志,2011,28(4):26-30]
[15]Zhang Y G,Luo Q S,Zhong M C.Studies on the structure of testis and spermatozoon in Leiocassis longirostris[J].Acta Hydrobiologica Sinica,1993,17(3):246-251[张耀光,罗泉笙,钟明超.长吻鮠精巢及精子结构的研究.水生生物学报,1993,17(3):246-251]
[16]You Y L,Lin D J.The ultrastructure of spermatozoon of the teleost Pseudobagrus fulvidraco[J].Acta Biologiae Experimentalis Sinica,1996,29(3):235-245[尤永隆,林丹军.黄颡鱼(Pseudobagrus fulvidraco)精子的超微结构.实验生物学报,1996,29(3):235-245]
[17]Gwo J C,Kuo M C,Chiu J Y,et al.Ultrastructure of Pagrus major and Rhabdosargus sarba spermatozoa(Perciformes:Sparidae:Sparinae)[J].Tissue and Cell,2004,36(2):141-147
[18]Gwo J C,Yang W T,Kuo M C,et al.Spermatozoal ultrastructures of two marine perciform teleost fishes,the goatfish,Paraupeneus spilurus(Mullidae)and the rabbitfish,Siganus fuscescens(Siganidae)from Taiwan[J].Tissue and Cell,2004,36(1):63-69
[19]Gwo J C,Chiu J Y,Lin C Y,et al.Spermatozoal ultrastructure of four Sparidae fishes:Acanthopagrus berda,Acanthopagrus australis,Lagodon rhomboids and Archosargus probatocephus[J].Tissue and Cell,2005,37(2):109-115
[20]Linhart O,Kudo S.Surface ultrastructure of paddlefish eggs before and after fertilization[J].Journal of Fish Biology,1997,51(3):573-582
[21]Xu Y,Xiong Q M.Cytological study of the fertilization of Acipenser sinensis Gray[J].Acta Zoologica Sinica,1990,36(3):275-278[许雁,熊全沬.中华鲟受精细胞学研究.动物学报,1990,36(3):275-279]
[22]Sun J J,Guo Y G,Li G F,et al.An observation on the early fertilization of Squaliobarbus curriculus by scanning electron microscope[J].Journal of Fishery Sciences of China,2006,13(5):740-744[孙际佳,郭云贵,李桂峰,等.赤眼鳟精子入卵的扫描电镜观察.中国水产科学,2006,13(5):740-744]
[23]Kudo S,Linhart O,Billard R.Ultrastructural studies of sperm penetration in the egg of the European catfish,Silurus glanis[J].Aquatic Living Resources,1994,7(2):93-98
[24]Fu C J,Liu X T,Shan J,et al.Cytological study on maturity of egg in Ctenopharyngodon idllus[J].Freshwater Fisheries,1981,(5):5-8[傅朝君,刘宪亭,单健,等.草鱼卵球成熟细胞学的研究.淡水渔业,1981,(5):5-8]
[25]Huang Y S.Cytological study on fertilization of egg in Tilapia nilotica[J].Acta Zoologica Sinica,1993,39(1):19-22[黄永松.尼罗罗非鱼卵母细胞受精细胞学研究.动物学报,1993,39(1):19-22]
[26]Wang R X,Zhang Y R,Fu C S,et al.The observation on the initial fertilization of freshwater bream by electronic scanning microscope[J].Journal of Fisheries of China,1982,6(4):313-320[王瑞霞,张毓人,傅仓生,等.鲂鱼受精早期精子入卵的扫描电子显微镜观察.水产学报,1982,6(4):313-320]
[27]Li P,Liu X L,Li Y,et al.Cytological study on fertilization process of Myxocyprinus asiaticus[J].Journal of Fishery Sciences of China,2013,20(2):250-259[李萍,刘晓蕾,李勇,等.胭脂鱼受精过程的细胞学观察.中国水产科学,2013,20(2):250-259]
[28]Shen Q Z,Wu K Z,Cai Z Y.Dynamical action of sperm penetration into the egg of Misgurnus anguillicaudatus[J].Zoological Research,1990,11(3):179-184[沈其璋,吴坤璋,蔡振岩.泥鳅精子入卵的动力作用.动物学研究,1990,11(3):179-184]
[29]Gan G M.Studies on Fertilization Biology of Hemibarbus labeo[D].Southwest University,2003[甘光明.唇?受精生物学研究.西南大学,2003]
[30]Wang Y M.Studies on the Fertilization Biology of Gobiocypris rarus[D].Southwest University,2011[王永明.稀有逗鲫受精生物学研究.西南大学,2011]
[31]Yao J H,Zhou P F.Scanning electron microscopic observation on the ultrastructure of the chorion and the process of insemination of Paramisgurnus dabryanus[J].Journal of Shanghai Fisheries University,1998,7(1):65-68[姚纪花,周平凡.大鳞副泥鳅卵子壳膜结构与授精过程的扫描电镜观察.上海水产大学学报,1998,7(1):65-68]
[32]Xu R W.An observation on the lifetime of loach M.anguillicaudatus spermatozoa[J].Journal of Zhejiang College of Fisheries(Natural Science Edition),1992,11(1):17[徐如卫.泥鳅Misgurnus anguillicaudatus Cantor精子寿命的初步观察.浙江海洋学院学报(自然科学版),1992,11(1):17]
[33]Zhang T,Liu L,Zhang J M,et al.Ultrastructure of spermatozoa and fertilized eggs of Anguilla japonica and observation on the fertilization process[J].Acta Hydrobiologica Sinica,2010,34(4):769-778[张涛,柳凌,张洁明,等.日本鳗鲡精卵的超微结构以及受精过程观察.水生生物学报,2010,34(4):769-778]