SCP3作为栉孔扇贝初级精母细胞标记分子的研究
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中国海洋大学,中国海洋大学,中国海洋大学,中国海洋大学,中国海洋大学,中国海洋大学

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Q786

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国家“八六三”高技术研究发展计划(2012AA10A402)


A potential role of SCP3 as a molecular marker of primary spermatocytes in Chlamys farreri
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Ocean University of China,Ocean University of China,Ocean University of China,Ocean University of China,Ocean University of China,Ocean University of China

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    摘要:

    为检验SCP3在无脊椎动物中是否可以标记特定的生殖细胞,实验利用RACE技术克隆了栉孔扇贝SCP3(Cf-SCP3)的全长cDNA序列,结果显示,其长度为1 033 bp,开放阅读框为726 bp,编码241个氨基酸。推导的氨基酸序列中包含SCP3保守的Cor1结构域和卷曲螺旋结构域。制备了Cf-SCP3地高辛标记的RNA探针和Cf-SCP3重组蛋白的多克隆抗体,采用原位杂交和免疫组织化学技术检测其细胞学定位,结果显示Cf-SCP3转录本和Cf-SCP3蛋白均特异性地定位在栉孔扇贝的初级精母细胞和未受精卵中,在精巢的其他类型细胞和卵巢中均未见表达信号。研究表明,Cf-SCP3蛋白可能与脊椎动物的SCP3蛋白一样,作为联会复合体的组成成分参与栉孔扇贝的配子发生,同时可以作为一个栉孔扇贝初级精母细胞的分子标记应用到体外诱导生殖细胞分化的研究。

    Abstract:

    Synaptonemal complex protein 3 (SCP3) is a meiosis-specific component of the synaptonemal complex essential for gametogenesis. In vertebrates, SCP3 protein is specifically localized in primary spermatocytes and many researchers have used it as a molecular marker to identify and separate primary spermatocytes from all types of germ cells. To measure whether SCP3 can mark any specific germ cells in invertebrates, we cloned a 1 033 bp full-length cDNA of Chlamys farreri SCP3 (Cf-SCP3) that contains a 726 bp open reading frame encoding 241 amino acids with one conserved Cor1 domain and two coiled-coil domains. Then we synthesized the DIG-labeled RNA probe, expressed the Cf-SCP3 recombinant protein and generated the anti-Cf-SCP3 polyclonal antibody. In situ hybridization and immunohistochemistry observations showed that Cf-SCP3 transcripts and Cf-SCP3 protein were specifically expressed in primary spermatocytes and unfertilized eggs, while no expression was detected in any other spermatogenic cells or ovarian cells of C. farreri. Thus, it suggested that the Cf-SCP3 protein might participate in meiosis of gametogenesis as a component of the synaptonemal complex like that in vertebrates and could be used as a candidate molecular marker for identifying the primary spermatocytes in germ cells differentiation study of C. farreri.

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马晓世,杨丹丹,季爱昌,梁少帅,马晓茹,张志峰.SCP3作为栉孔扇贝初级精母细胞标记分子的研究[J].水产学报,2017,41(7):1096~1104

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  • 收稿日期:2016-03-17
  • 最后修改日期:2016-12-02
  • 录用日期:2017-03-28
  • 在线发布日期: 2017-07-24
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