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对虾卵巢发育过程中的比较蛋白质组学分析。

Comparative proteomic profiling during ovarian development of the shrimp Metapenaeus ensis.

机构信息

The Key Laboratory of Geriatrics, Beijing Hospital and Beijing Institute of Geriatrics, Ministry of Health, Da Hua Road, Beijing, 100730, China,

出版信息

Mol Biol Rep. 2014 Jan;41(1):519-28. doi: 10.1007/s11033-013-2887-6. Epub 2013 Dec 6.

Abstract

Two-dimensional electrophoresis and mass spectrometry were used to identify proteins that are differentially expressed during ovarian maturation in Metapenaeus ensis. 87 spots with consistently significant quantitative differences (≥ 1.5-fold for vol%) among stage I, III and V ovaries were chosen for MS/MS analysis. 45 spots were significantly matched to known proteins in the database (Mascot score >40). Half of them were down-regulated, in contrast to 9 out of 45 proteins that were up-regulated as ovarian maturation proceeded. Functionally, these identified proteins could be classified into five major groups, including cytoskeleton (11 %), metabolism (18 %), signal transduction (32 %), gene expression (14 %) and immune response (7 %). Among the differentially expressed reproduction-related proteins, the mRNA expression level of cellular retinoic acid/retinol binding protein in M. ensis (MeCRABP) during ovarian maturation was further characterized by quantitative real-time PCR. It was down-regulated during ovarian maturation. In situ hybridization further revealed that MeCRABP transcript was localized in ooplasm of previtellogenic oocytes but not in vitellogenic oocytes. These results demonstrate the application of proteomic analysis for identification of proteins involved in shrimp ovarian maturation and they provide new insights into ovarian development.

摘要

采用二维电泳和质谱技术鉴定了中国明对虾卵巢发育过程中差异表达的蛋白质。选择在 I 期、III 期和 V 期卵巢之间具有一致显著定量差异(体积%≥1.5 倍)的 87 个斑点进行 MS/MS 分析。45 个斑点与数据库中的已知蛋白质有显著匹配(Mascot 得分>40)。其中一半下调,而在 45 个上调的蛋白质中,有 9 个随着卵巢发育而上调。功能上,这些鉴定的蛋白质可以分为五个主要组,包括细胞骨架(11%)、代谢(18%)、信号转导(32%)、基因表达(14%)和免疫反应(7%)。在差异表达的与生殖相关的蛋白质中,中国明对虾细胞视黄酸/视黄醇结合蛋白(MeCRABP)的 mRNA 表达水平在卵巢发育过程中通过定量实时 PCR 进一步进行了特征描述。在卵巢发育过程中,其表达水平下调。原位杂交进一步揭示 MeCRABP 转录本定位于卵母细胞的卵质中,但不在卵黄发生卵母细胞中。这些结果表明蛋白质组学分析可用于鉴定参与虾类卵巢成熟的蛋白质,并为卵巢发育提供新的见解。

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