皱纹盘鲍与西氏鲍杂交受精过程荧光显微观察
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摘要
本研究采用荧光显微技术对皱纹盘鲍(Halilotis discus hannai)♀×西氏鲍(H.gigantea)♂的受精过程及受精卵的早期发育进行了观察,并采用FITC-anti-α-tubulin对精子入卵位置进行标记.结果显示西氏鲍的精子可正常激活皱纹盘鲍的卵子,精子进入卵子2-5 min后受精卵开始第一次卵裂,释放出第一极体.随着受精卵的发育卵子完成第二次减数分裂,排出第二极体,同时形成早期精卵原核.精卵原核膨胀并彼此融合,受精卵开始进行有丝分裂完成第一次卵裂.通过FITC标记的anti-α-tubulin蛋白免疫荧光观察,发现精子在入卵时会在卵膜表面,形成一个类似于受精孔的结构.
The current study was conducted on the fertilization process and the early development of fertilized eggs by the fluorescence microscopy technique.The position of sperm into the eggs was labeled by FITC-anti-α-tubulin.The results showed that sperm of H.gigantea penetrated H.discus hannai eggs.The first cleavage and the first polar body were released in after 2-5min of sperm-egg fusion.With the completion of the second meiotic division of the fertilized egg,the second polar body was released,forming the early male and female pronucleus.Then two pronucleus began to swell and started mutual fusion,and the fertilized eggs began to undergo mitosis to complete the first cleavage.The immunofluorescent observation with the FITC labeled anti-α-tubulin probe detected a pore structure similar to micropylar on the surface of egg membrane.
The current study was conducted on the fertilization process and the early development of fertilized eggs by the fluorescence microscopy technique.The position of sperm into the eggs was labeled by FITC-anti-α-tubulin.The results showed that sperm of H.gigantea penetrated H.discus hannai eggs.The first cleavage and the first polar body were released in after 2-5min of sperm-egg fusion.With the completion of the second meiotic division of the fertilized egg,the second polar body was released,forming the early male and female pronucleus.Then two pronucleus began to swell and started mutual fusion,and the fertilized eggs began to undergo mitosis to complete the first cleavage.The immunofluorescent observation with the FITC labeled anti-α-tubulin probe detected a pore structure similar to micropylar on the surface of egg membrane.
引文
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[18]骆轩,游伟伟,柯才焕,等.西氏鲍与盘鲍杂交育苗的初步研究[J].厦门大学学报(自然科学版),2006,45(5):602-605.
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[24]蔡明夷,柯才焕,游伟伟,等.杂色鲍♀×盘鲍♂杂交受精的细胞学研究[J].厦门大学学报(自然科学版),2007,46(2):239-243.
[25]Vacquier,V.D.,Carner,K.R.Stout,C.D.Species-Specific sequences of Abalone lysin,the sperm protein that creates a hole in the egg envelope[J].Proceedings of the National Academy of Sciences of the United States of America,1990,87(15):5792-5796.
[26]孙振兴,王如才.皱纹盘鲍卵的形态与超显微结构[J].中国水产科学,1997,4(1):68-74.
[27]朱蓝菲,蒋一珪.银鲫不同雌核发育系的生物学特性比较研究[J].水生生物学报,1993,17(2):112-120.
[28]骆轩.西氏鲍与皱纹盘鲍杂交的遗传基础研究[D].厦门:厦门大学,2005.
[29]Shaw,A.,Mcree,D.E.,Vacquier V.D.The crystal structure of lysin,a fertilization protein[J].Science,1993,262(5141):1864-1867.
[30]Lee,Y.H.,Vacquier,V.D.Evolution and systematics in Haliotidae(Mollusca:Gastropoda):Inferences from DNA sequences of sperm lysin[J].Marine Biology,1995.,124(2):267-278.
[31]毕克,包振民,黄晓婷,等.栉孔扇贝♀×华贵栉孔扇贝♂受精及早期胚胎发育过程的细胞学荧光显微观察[J].中国海洋大学学报(自然科学版),2005,35(2):283-286.
[32]Mozingo,N.M.,Vacquier,V.D.,Chandler,D.E.Structural features of the abalone egg extracellular matrix and its role in gamete interaction during fertilization[J].Molecular reproduction and development,1995,41(4):493-502.
[33]Lewis,C.A.,Talbot,C.F.,Vacquier,V.D.A protein from abalone sperm dissolves the egg vitelline layer by a nonenzymatic mechanism[J].Developmental biology,1982,92(1):227-239.
[34]Raven,C.P.Morphogenesis:The analysis of molluscan development[M].Oxford:Pergamon Press,1966.
[2]王仁波,范家春.红鲍人工育苗及其与皱纹盘鲍杂交试验的初步研究[J].大连水产学院学报,1999,14(3):64-66.
[3]燕敬平,张榭令.日本盘鲍与皱纹盘鲍杂交育种技术研究[J].海洋水产研究,1999,20(1):35-39.
[4]Sun Z.,Chang L.,Song Z.Effects of hybridization between Haliotis discus hannaiand Haliotis discus[J].Fisheries Science,2005,24:1-3.
[5]柯才焕,田越,周时强.杂色鲍与皱纹盘鲍,盘鲍杂交的初步研究[J].海洋科学,2000,24(11):39-41.
[6]蔡明夷,柯才焕,游伟伟,等.杂色鲍♀×盘鲍♂杂交受精的细胞学研究[J].厦门大学学报(自然科学版),2007,46(2):239-243.
[7]Cai M.Y.,Ke C.H.,Wang G.Z.,et al.Influence factors on fertilization rate in laboratory hybridization between Haliotisdiversicolor and H.discus discus[J].Journal of Fisheries of China,2006,13:230-236.
[8]Cai M.Y.,Ke C.H.,Luo X.A.,et al.Karyological studies of the hybrid Larvae of Haliotisdisversicolor supertextafemale and Haliotis discus discusmale[J].Journal of Shellfish Research,2010,29(3):735-740.
[9]孙振兴,宋志乐.日本大鲍与皱纹盘鲍杂交的研究[J].齐鲁渔业,2001,18(3):25-27.
[10]Sun Z.,Song Z.,Zhen Z.,et al.Hybridization between two species abalone and growth of their F1 hybrid progeny[C].proceedings of the 5th International Abalone Symposium,China:Qingdao,F,2003.
[11]郑升阳.日本西氏鲍与皱纹盘鲍杂交试验初报[J].福建农业学报,2006,21(3):296-298.
[12]骆轩,游伟伟,柯才焕,等.西氏鲍与盘鲍杂交育苗的初步研究[J].厦门大学学报(自然科学版),2006,45(2):602-605.
[13]Luo X.,Ke C.H.,You W.W.,et al.Factors affecting the fertilization Success in laboratory hybridization between Haliotis discus hannai and Haliotisgigantea[J].Journal of Shellfish Research,2010,29(3):621-625.
[14]Luo X.,Ke C.H.,You W.W.Heterosis analysis on interspcific hybrids between Haliotis discus hannai and H.gigantea[J].Journal of Shellfish Research,2012,31(1):316-316.
[15]范飞龙.绿鲍的引种及与皱纹盘鲍的种间杂交研究[D].厦门:厦门大学,2012.
[16]张国范.海洋贝类遗传育种研究20年[J].厦门大学学报(自然科学版),2006,45(2):190-194.
[17]You W.W.,Ke C.H.,Luo X.,et al.Growth and survival of three small abalone Haliotisdiversicolor populations and their reciprocal crosses[J].Aquaculture Research,2009,40(13):1474-1480.
[18]骆轩,游伟伟,柯才焕,等.西氏鲍与盘鲍杂交育苗的初步研究[J].厦门大学学报(自然科学版),2006,45(5):602-605.
[19]Wang,H.S.,Luo X.,You W.W.,et al.Cytogenetic analysis and chromosomal characteristics of the polymorphic 18S r DNA of Haliotis discus hannai from Fujian,China[J].PLOS ONE,2015,10(2):e0113816.
[20]Chen,D.Y.,Frank,J.L.Sperm nuclear dispersion coordinate with meiotic maturation in fertilized Spisula solidissima eggs[J].Developmental Biology,1983,99(1):217-224.
[21]Longo,F.J.,Mathews,L.,Hedgecock,D.Morphogenesis of maternal and paternal genomes in fertilized oyster eggs(Crassostreagigas):effects of cytochalasin B at different periods during meiotic maturation[J].Biological Bulletin,1993,185(2):197-214.
[22]楼允东,郑德崇.组织胚胎学[M].北京:农业出版社,1981.
[23]孙振兴,谢嘉琳.皱纹盘鲍受精过程的电镜观察[J].动物学研究,1997,18(3):253-257.
[24]蔡明夷,柯才焕,游伟伟,等.杂色鲍♀×盘鲍♂杂交受精的细胞学研究[J].厦门大学学报(自然科学版),2007,46(2):239-243.
[25]Vacquier,V.D.,Carner,K.R.Stout,C.D.Species-Specific sequences of Abalone lysin,the sperm protein that creates a hole in the egg envelope[J].Proceedings of the National Academy of Sciences of the United States of America,1990,87(15):5792-5796.
[26]孙振兴,王如才.皱纹盘鲍卵的形态与超显微结构[J].中国水产科学,1997,4(1):68-74.
[27]朱蓝菲,蒋一珪.银鲫不同雌核发育系的生物学特性比较研究[J].水生生物学报,1993,17(2):112-120.
[28]骆轩.西氏鲍与皱纹盘鲍杂交的遗传基础研究[D].厦门:厦门大学,2005.
[29]Shaw,A.,Mcree,D.E.,Vacquier V.D.The crystal structure of lysin,a fertilization protein[J].Science,1993,262(5141):1864-1867.
[30]Lee,Y.H.,Vacquier,V.D.Evolution and systematics in Haliotidae(Mollusca:Gastropoda):Inferences from DNA sequences of sperm lysin[J].Marine Biology,1995.,124(2):267-278.
[31]毕克,包振民,黄晓婷,等.栉孔扇贝♀×华贵栉孔扇贝♂受精及早期胚胎发育过程的细胞学荧光显微观察[J].中国海洋大学学报(自然科学版),2005,35(2):283-286.
[32]Mozingo,N.M.,Vacquier,V.D.,Chandler,D.E.Structural features of the abalone egg extracellular matrix and its role in gamete interaction during fertilization[J].Molecular reproduction and development,1995,41(4):493-502.
[33]Lewis,C.A.,Talbot,C.F.,Vacquier,V.D.A protein from abalone sperm dissolves the egg vitelline layer by a nonenzymatic mechanism[J].Developmental biology,1982,92(1):227-239.
[34]Raven,C.P.Morphogenesis:The analysis of molluscan development[M].Oxford:Pergamon Press,1966.