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腹足类胚胎发育过程中的蛋白表达。

Protein expression during the embryonic development of a gastropod.

机构信息

Department of Biology, Hong Kong Baptist University, Hong Kong, P R China.

出版信息

Proteomics. 2010 Jul;10(14):2701-11. doi: 10.1002/pmic.200900846.

Abstract

Despite the potential use of gastropod embryos in basic and applied research, little is known about their protein expression. We examined, for the first time, changes in proteomic profile during embryonic development of Pomacea canaliculata from an embryo without a shell (stage II) to an embryo with a fully formed shell (stage III) to understand the roles that proteins play in critical developmental events, such as the formation of shell, operculum and heart, and the differentiation of head and foot. To analyze protein expression during development, we used 2-DE to detect, MS to analyze, and de novo peptide sequencing followed by MS-BLAST to identify the proteins. The de novo cross-species protein identification method was adopted because of a lack of genomic and proteomic data in the whole class of Gastropoda. 2-DE detected approximately 700 protein spots. Among the 125 spots that were abundant, 52% were identified, a marked improvement over the conventional direct MS-BLAST method. These proteins function in perivitelline fluid utilization, shell formation, protein synthesis and folding, and cell cycle and cell fate determination, providing evidence to support that this embryonic period is a period of dynamic protein synthesis and metabolism. The data shall provide a basis for further studies of how gastropod embryos respond to natural and human-induced changes in the environment.

摘要

尽管腹足类胚胎在基础和应用研究中有潜在的用途,但对其蛋白质表达的了解甚少。我们首次研究了在Pomacea canaliculata胚胎发育过程中蛋白质组图谱的变化,从没有壳的胚胎(II 期)到完全形成壳的胚胎(III 期),以了解蛋白质在壳、瓣和心脏形成以及头足分化等关键发育事件中的作用。为了分析发育过程中的蛋白质表达,我们使用 2-DE 进行检测、MS 进行分析、从头测序肽段并通过 MS-BLAST 进行鉴定。由于腹足纲的整个类群缺乏基因组和蛋白质组数据,因此采用了从头跨物种蛋白质鉴定方法。2-DE 检测到大约 700 个蛋白质斑点。在 125 个丰富的斑点中,有 52%被鉴定出来,这比传统的直接 MS-BLAST 方法有了显著的改进。这些蛋白质参与卵黄周围液体的利用、壳的形成、蛋白质的合成和折叠以及细胞周期和细胞命运的决定,为这一胚胎期是一个动态蛋白质合成和代谢的时期提供了证据。这些数据将为进一步研究腹足类胚胎如何应对自然和人为引起的环境变化提供基础。

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