Identification of male-specific amh duplication, sexually differentially expressed genes and microRNAs at early embryonic development of Nile tilapia (Oreochromis niloticus).
Identification of male-specific amh duplication, sexually differentially expressed genes and microRNAs at early embryonic development of Nile tilapia (Oreochromis niloticus).
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DOI：
10.1186/1471-2164-15-774
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发表时间：
2014-09-09
期刊：
影响因子：
4.4
通讯作者：
Ron M
中科院分区：
文献类型：
--
作者：
Eshel O;Shirak A;Dor L;Band M;Zak T;Markovich-Gordon M;Chalifa-Caspi V;Feldmesser E;Weller JI;Seroussi E;Hulata G;Ron M
基因和环境对尼罗罗非鱼性别决定的可能影响表明，性别决定应被视为一个复杂的性状。罗非鱼性别决定基因的检测具有重要的科学意义和经济价值。主要目的是检测在早期胚胎发育中性别差异表达的基因和microRNA。将尼罗罗非鱼XX雌鱼分别与性别逆转的ΔXX雄鱼和基因改造的YY“超级雄鱼”人工受精，分别获得全雌和全雄胚胎。将2、5和9dpf的全雌性和全雄性胚胎池的RNA用作定制Agilent eArray杂交和下一代测序的模板。通过p < 0.05的错误发现率鉴定出性别间差异表达的59个基因。男性中最高表达的基因是amh和tspan 8，女性中最高表达的基因是cr/20β-hsd、gpa 33、rtn 4 ipl和zp 3（p < 1 × 10−9）。在胚胎和性腺中使用qPCR验证基因表达表明tspan 8、gpa 33、cr/20β-hsd和amh的拷贝数变化。amh的测序鉴定了该基因的男性特异性重复，表示为amhy，与amh的序列不同之处在于外显子VII上的233 bp缺失，因此缺乏编码与转化生长因子β受体（TGF-β结构域）结合的蛋白基序的能力。amh和amhy在作图家系中的分离与LG 23上的SD连锁标记完全一致，该标记表示SD的QTL。我们在罗非鱼胚胎中发现了831个microRNA，其中9个具有性二态表达模式，假发现率为p < 0.05。在雄性中上调的microRNA pma-mir-4585的特征在于所有六个预测的靶基因，包括在雄性中下调的cr/20β-hsd。本研究首次在罗非鱼胚胎发育的极早期阶段，即从2 dpf开始，发现了性别差异表达的基因和microRNA。具有性别差异表达模式的基因因拷贝数变异而富集。鉴定了一种新的雄性特异性amh重复，称为amhy，缺乏TGF-β结构域，并定位到SD的LG 23上的QTL区域，从而表明其在SD中的潜在作用。本文的在线版本（doi：10.1186/1471-2164-15-774）包含补充材料，可供授权用户使用。
The probable influence of genes and the environment on sex determination in Nile tilapia suggests that it should be regarded as a complex trait. Detection of sex determination genes in tilapia has both scientific and commercial importance. The main objective was to detect genes and microRNAs that were differentially expressed by gender in early embryonic development. Artificial fertilization of Oreochromis niloticus XX females with either sex-reversed ΔXX males or genetically-modified YY ‘supermales’ resulted in all-female and all-male embryos, respectively. RNA of pools of all-female and all-male embryos at 2, 5 and 9 dpf were used as template for a custom Agilent eArray hybridization and next generation sequencing. Fifty-nine genes differentially expressed between genders were identified by a false discovery rate of p < 0.05. The most overexpressed genes were amh and tspan8 in males, and cr/20β-hsd, gpa33, rtn4ipl and zp3 in females (p < 1 × 10−9). Validation of gene expression using qPCR in embryos and gonads indicated copy number variation in tspan8, gpa33, cr/20β-hsd and amh. Sequencing of amh identified a male-specific duplication of this gene, denoted amhy, differing from the sequence of amh by a 233 bp deletion on exonVII, hence lacking the capability to encode the protein motif that binds to the transforming growth factor beta receptor (TGF-β domain). amh and amhy segregated in the mapping family in full concordance with SD-linked marker on LG23 signifying the QTL for SD. We discovered 831 microRNAs in tilapia embryos of which nine had sexually dimorphic expression patterns by a false discovery rate of p < 0.05. An up-regulated microRNA in males, pma-mir-4585, was characterized with all six predicted target genes including cr/20β-hsd, down-regulated in males. This study reports the first discovery of sexually differentially expressed genes and microRNAs at a very early stage of tilapia embryonic development, i.e. from 2 dpf. Genes with sexually differential expression patterns are enriched for copy number variation. A novel male-specific duplication of amh, denoted amhy, lacking the TGF-β domain was identified and mapped to the QTL region on LG23 for SD, thus indicating its potential role in SD. The online version of this article (doi:10.1186/1471-2164-15-774) contains supplementary material, which is available to authorized users.
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罗非鱼（Oreochromis niloticus）中的TOL2介导的转基因。
影响因子：
4.5
作者：
Fujimura, Koji;Kocher, Thomas D.
通讯作者：
Kocher, Thomas D.
影响因子：
--
作者：
Garcia-Ortiz JE;Pelosi E;Omari S;Nedorezov T;Piao Y;Karmazin J;Uda M;Cao A;Cole SW;Forabosco A;Schlessinger D;Ottolenghi C
通讯作者：
Ottolenghi C
影响因子：
2.7
作者：
Golan, Matan;Levavi-Sivan, Berta
通讯作者：
Levavi-Sivan, Berta
影响因子：
4.8
作者：
Chen, Ting;Li, Meng;Li, Lin
通讯作者：
Li, Lin
影响因子：
7.6
作者：
Cryns, Kim;Shamir, Alon;Moechars, Dieder
通讯作者：
Moechars, Dieder