Male-specific Dmrt1 is a candidate sex determination gene in spotted scat (Scatophagus argus)
Male-specific Dmrt1 is a candidate sex determination gene in spotted scat (Scatophagus argus)
复制标题
雄性特异性 Dmrt1 是斑点粪便 (Scatophagus argus) 中的候选性别决定基因
DOI：
10.1016/j.aquaculture.2018.06.009
复制
发表时间：
2018-10-01
期刊：
影响因子：
4.5
通讯作者：
Li, Guang-Li
中科院分区：
文献类型：
--
作者：
Mustapha, Umar Farouk;Jiang, Dong-Neng;Li, Guang-Li
D 乌鳢是一种养殖的海洋硬骨鱼，雌性比雄性生长更快、体型更大，因此克隆其性别决定主基因（SD）将有助于其在水产养殖中的性别控制。高度保守的 Dmrt1（双性和 mab-3 相关转录因子 1）基因编码锌指样 DNA 结合基序（DM 结构域），在脊椎动物的性别决定和分化中发挥着关键作用。在这里，两对性别特异性引物 Dmrt3-Marker-F/R 和 Dmrt1-Marker-F/R 分别是从乌鳢中的 Dmrt3 和 Dmrt1 转录本开发的。 Dmrt3-Marker-F/R扩增的DNA上第151和166 bp的单核苷酸多态性(SNP)在女性中均为纯合CC GG基因型，在男性中均为杂合CG GA基因型。 Dmrt1-Marker-F/R 只能在雄性中扩增 3.2 kb 的片段。这两种标记均在中国 5 个群体的 246 只成年乌鳢（113 只雄性和 133 只雌性）中得到证实。这些结果表明乌鳢的性别是由性染色体控制的（XX雌性，XY雄性）。 Dmrt3-Marker-F/R 扩增的 DNA 166 bp 位置处的核苷酸被发现是 Dmrt3 中的供体剪接位点突变 (c.400+ 1G > A)，产生截短的同种型。截短的Dmrt3与男性相关，后被命名为Dmrt3(Delta)-Y，其等位基因是X染色体上的正常Dmrt3。 Dmrt1-Marker-F/R 扩增的 DNA 是正常 Dmrt1 外显子 2 和外显子 3 之间的片段，这意味着 Dmrt1 是男性特异性的并且位于 Y 染色体上。研究发现男性和女性体内均存在截短的 Dmrt1 (Dmrt1b)。 Dmrt1b与Dmrt1高度相似，表明Dmrt1b是位于X染色体上的Dmrt1的等位基因。 RT-PCR显示Dmrt1在睾丸中高表达，而在卵巢中不表达。 Dmrt3(Delta)-Y仅在一些睾丸中微弱表达，而在卵巢中不表达。睾丸特异性表达的Dmrt1是乌鳢Y染色体上的候选SD基因。
D The Scatophagus argus is a cultured marine teleost fish in which females grow faster and larger than males, therefore cloning the master sex-determination (SD) gene will be helpful for its sex control in aquaculture. A well-conserved Dmrt1 (doublesex-and mab-3-related transcription factor-1) gene, encoding zinc finger-like DNA binding motif (DM-domain) plays a critical role in sex determination and differentiation in vertebrates. Here, two pairs of sex-specific primers, Dmrt3-Marker-F/R and Dmrt1-Marker-F/R were developed from Dmrt3 and Dmrt1 transcripts in S. argus respectively. The single-nucleotide polymorphisms (SNP) at position - 151 and 166 bp on the DNA amplified by Dmrt3-Marker-F/R are all homozygous CC GG genotype in females and heterozygous CG GA genotype in the males. The Dmrt1-Marker-F/R could only amplify a 3.2 kb fragment in males. Both Markers were confirmed in 246 adult S. argus (113 males and 133 females) from five populations in China. These results indicate that the sexes of S. argus are controlled by sex chromosomes (XX females, XY males). The nucleotide at position 166 bp of the DNA amplified by Dmrt3-Marker-F/R is found to be a donor splice site mutation (c.400+ 1G > A) in Dmrt3 which generates a truncated isoform. The truncated Dmrt3 is linked to male sex and thereafter named as Dmrt3(Delta)-Y, and its allele is the normal Dmrt3 on the X chromosome. The DNA amplified by Dmrt1-Marker-F/R is a fragment between exon 2 and exon 3 of a normal Dmrt1 implying that Dmrt1 is male-specific and located on the Y chromosome. A truncated Dmrt1 (Dmrt1b) was found to exist in both males and females. The Dmrt1b is with high similarity with Dmrt1 indicating Dmrt1b is the allele of Dmrt1 located on the X chromosome. RT-PCR revealed that Dmrt1 is highly expressed in the testes, while it is not expressed in the ovaries. Dmrt3(Delta)-Y is only expressed weakly in some of the testes, while it is not expressed in the ovaries. The testicular specifically expressed Dmrt1 is the candidate SD gene on the Y chromosome in S. argus.