Proteome analysis of early lineage specification in bovine embryos
Proteome analysis of early lineage specification in bovine embryos
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DOI：
10.1002/pmic.201400251
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发表时间：
2015-02-01
期刊：
影响因子：
3.4
通讯作者：
Arnold, Georg J.
中科院分区：
文献类型：
--
作者：
Demant, Myriam;Deutsch, Daniela R.;Arnold, Georg J.
在哺乳动物胚胎发育过程中，受精卵在输卵管中经历胚胎分裂，并在桑葚胚阶段到达子宫，此时发生压实和早期谱系规范。为了增加对胚胎蛋白库相关变化的了解，我们使用基于 iTRAQ 的方法对体外产生的牛桑葚和胚泡（六个生物重复）进行了全面的蛋白质组分析。总共鉴定了 560 种蛋白质，其中 502 种进行了定量。桑葚和囊胚中140种蛋白质的丰度存在显着差异，其中核磷蛋白（NPM1）、真核翻译起始因子5A-1（EIF5A）、活化蛋白激酶C 1 受体（GNB2L1/RACK1）和膜联蛋白A6（ANXA6）增加，谷胱甘肽S-转移酶mu 3（GSTM3）、过氧化还原蛋白2 (PRDX2) 和醛酮还原酶家族 1 成员 B1 (AKR1B1) 在囊胚中丰度降低。 73% 的丰度改变的蛋白质增加，反映了这一时期翻译活动的增加。参与翻译机制和 ATP 合成的蛋白质丰度的增加进一步支持了这一点。此外，互补的 2D 饱和 DIGE 分析可检测蛋白质亚型，例如GSTM3 和 PRDX2 与哺乳动物发育的这一时期相关，并示例性地验证了 iTRAQ 方法的结果。总之，我们对牛桑葚和胚泡的系统差异蛋白质组分析揭示了牛胚胎发生过程中早期谱系规范和分化事件的新分子相关性。
During mammalian embryo development, the zygote undergoes embryonic cleavage in the oviduct and reaches the uterus at the morula stage, when compaction and early lineage specification take place. To increase knowledge about the associated changes of the embryonic protein repertoire, we performed a comprehensive proteomic analysis of in vitro produced bovine morulae and blastocysts (six biological replicates), using an iTRAQ-based approach. A total of 560 proteins were identified of which 502 were quantified. The abundance of 140 proteins was significantly different between morulae and blastocysts, among them nucleophosmin (NPM1), eukaryotic translation initiation factor 5A-1 (EIF5A), receptor of activated protein kinase C 1 (GNB2L1/RACK1), and annexin A6 (ANXA6) with increased, and glutathione S-transferase mu 3 (GSTM3), peroxiredoxin 2 (PRDX2), and aldo-keto reductase family 1 member B1 (AKR1B1) with decreased abundance in blastocysts. Seventy-three percent of abundance altered proteins increased, reflecting an increase of translation activity in this period. This is further supported by an increase in the abundance of proteins involved in the translation machinery and the synthesis of ATP. Additionally, a complementary 2D saturation DIGE analysis led to the detection of protein isoforms, e.g. of GSTM3 and PRDX2, relevant for this period of mammalian development, and exemplarily verified the results of the iTRAQ approach. In summary, our systematic differential proteome analysis of bovine morulae and blastocysts revealed new molecular correlates of early lineage specification and differentiation events during bovine embryogenesis.