管角螺受精过程的荧光显微观察及保育卵形成分析
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宁波大学海洋学院,宁波大学海洋学院,宁波大学海洋学院,宁波大学海洋学院,宁波大学海洋学院

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国家自然科学基金(41206113);浙江省教育厅科技计划(Y201224623);“十二五”农村领域国家科技计划课题(2011BAD13B0903);浙江海洋高效健康养殖协同创新中心


Cytological observation of fertilization and analyse of the nurse eggs in Hemifusus tuba (Gmelin)
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School of Marine Sciences,Ningbo University,School of Marine Sciences,Ningbo University,School of Marine Sciences,Ningbo University,School of Marine Sciences,Ningbo University,School of Marine Sciences,Ningbo University

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    摘要:

    为了研究管角螺受精卵在受精和早期卵裂过程中的核相变化以及保育卵的形成,实验通过Hoechst 33258染色、荧光显微观察方法,对管角螺受精和早期卵裂过程中核行为的细胞学变化进行了详细观察,并对其胚胎发育过程中的数量变化进行统计分析。结果显示,管角螺的成熟未受精卵呈卵圆形,平均卵径(0.33±0.03)mm,核相处于第一次成熟分裂中期,在水温为(26±2)℃条件下,1 h精子入卵,2~3 h受精卵先后排出第1、第2极体,完成第一次和第二次成熟分裂,6 h雌、雄性原核形成,靠近融合,第一次卵裂形成2细胞,12~24 h时2细胞继续分裂生成4细胞。研究表明,管角螺受精卵在成熟分裂和卵裂过程中表现出发育的不同步性,同个卵荚内多个发育阶段共存,受精率为43.14%,而受精卵正常发育概率仅5.70%。卵荚内多数卵子不受精或者受精卵停止发育,成为营养卵(保育卵),为正常可育卵消化利用,这两种来源的保育卵比率约为3:2。管角螺胚胎发育过程中,1~6 d内胚胎数减少明显,担轮幼虫发育率为0.48%,之后数量稳定,直至稚螺出膜。

    Abstract:

    The cytological changes of nuclear behavior during the process of fertilization and early cleavage of Hemifusus tuba (Gmelin) were continually observed by fluorescent microscope. The numbers of eggs and embryos were also calculated for the whole embryonic development stages. The results showed that mature unfertilized eggs of H. tuba were oval and the average diameter was 0.33 ± 0.03 mm. The nuclear phase was at the metaphase of the first maturation division. At water temperature of 27-28℃, sperm penetrated into cytoplasm of egg 1h after fertilization. The fertilized eggs released the first polar body and the second polar body at 2~3 h. The female pronuclei and male pronuclei formed at 6 h, followed by the first cleavage. The second cleavage happened at 12-24 h, formed 4-cells. The study showed that the zygotes of H. tuba were asynchronous during meiosis and cleavage. In total, the fertilization ratio was about 43.14%, but the zygotes which could develop to 4-cell stage were only 5.70%. In a capsule, many eggs were not fertilized, or stopped developing, and both of them were inferred as the sources of nurse eggs which were consumed by developing embryos. The ratio of eggs unfertilized to eggs that stopped developing was 3 to 2. During the embryonic development, the number of eggs reduced significantly during the 1st to 6th day, only 0.48% of eggs developed to trochophore stage, and this ratio maintained until the juveniles left the capsules for natural environment.

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杨泽琴,李荣华,母昌考,宋微微,王春琳.管角螺受精过程的荧光显微观察及保育卵形成分析[J].水产学报,2016,40(11):1674~1682

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  • 收稿日期:2016-01-20
  • 最后修改日期:2016-07-22
  • 录用日期:2016-11-15
  • 在线发布日期: 2016-11-25
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