Identification and characterization of a Masculinizer (Masc) gene involved in sex differentiation in Artemia
Identification and characterization of a Masculinizer (Masc) gene involved in sex differentiation in Artemia
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参与卤虫性别分化的男性化基因 (Masc) 的鉴定和表征
DOI：
10.1016/j.gene.2017.03.010
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发表时间：
2017-05-30
期刊：
影响因子：
3.5
通讯作者：
Yang, Wei-Jun
中科院分区：
文献类型：
--
作者：
Li, Dong-Rui;Ye, Hui-Li;Yang, Wei-Jun
无脊椎动物等相对原始的动物的性别大多是由环境因素和染色体倍性决定的。异形染色体也可能发挥重要作用，如鳞翅目动物的 ZW 系统。然而，这些不同的性别决定系统的机制在很大程度上仍然不明确。在本研究中，在甲壳类动物Artemia franciscana Kellogg 1906中鉴定出Masculinizer基因（Ar-Masc）。序列分析显示，Ar-Masc的1140 bp全长开放阅读框编码一个380-aa的蛋白质，其中包含两个CCCH型锌指结构域，与家蚕Bombyx mori中特征的MASC蛋白具有高度的共享同一性，已确定该蛋白参与其中用于产生雄性特异性剪接变体。此外，尽管在有性贫血和孤雌性贫血的几乎所有阶段都可以检测到 Ar-Masc，但这两种生殖模式之间的表达存在显着差异。首先，qRT-PCR和Western blot分析表明，在整个孵化过程中，有性无节幼体中Ar-Masc mRNA和蛋白质的水平远高于孤雌无节幼体。其次，有性丰年虫和孤雌丰年虫的Ar-Masc水平均随胚胎发育阶段而降低，而有性丰年虫的Ar-Masc表达量较孤雌丰年虫相对较高且稳定。第三，免疫荧光分析确定有性个体在胚胎发育过程中比孤雌个体具有更高水平的Ar-MASC蛋白。最后，对dsRNA的RNA干扰表明，有性毛茛Ar-Masc基因沉默导致子代的雌雄比为2.19:1。这些数据表明 Ar-Masc 参与了 A. franciscana 的性别决定过程，并为有性生物性别决定的进化提供了见解。 (C) 2017 Elsevier B.V. 保留所有权利。
The sex of relatively primitive animals such as invertebrates is mostly determined by environmental factors and chromosbme ploidy. Heteromorphic chromosomes may also play an important role, as in the ZW system in lepidopterans. However, the mechanisms of these various,sex determination systems are still largely undefined. In the present study, a Masculinizer gene (Ar-Masc) was identified in the crustacean Artemia franciscana Kellogg 1906. Sequence analysis revealed that the 1140-bp full-length open reading frame of Ar-Masc encodes a 380-aa protein containing two CCCH-type zinc finger domains having a high degree of shared identities with the MASC protein characterized in the silkworm Bombyx mori, which has been determined to participate in the production of male-specific splice variants. Furthermore, although Ar-Masc could be detected in almost all stages in both sexual and parthenogenetic Anemia, there were significant variations in expression between these two reproductive modes. Firstly, qRT-PCR and Western blot analysis showed that levels of both Ar-Masc mRNA and protein in sexual nauplii were much higher than in parthenogenetic nauplii throughout the hatching process. Secondly, both sexual and parthenogenetic Artemia had decreased levels of Ar-Masc along with the embryonic developmental stages, while the sexual ones had a relatively higher and more stable expression than those of parthenogenetic ones. Thirdly, immunofluorescence analysis determined that sexual individuals had higher levels of Ar-MASC protein than parthenogenetic individuals during embryonic development. Lastly, RNA interference with dsRNA showed that gene silencing of Ar-Masc in sexual A. franciscana caused the female-male ratio of progeny to be 2.19:1. These data suggest that Ar-Masc participates in the process of sex determination in A. franciscana, and provide insight into the evolution of sex determination in sexual organisms. (C) 2017 Elsevier B.V. All rights reserved.