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基于 iTRAQ 的太平洋牡蛎受精卵低盐响应的蛋白质组谱分析。

iTRAQ-based proteome profiling of hyposaline responses in zygotes of the Pacific oyster Crassostrea gigas.

机构信息

Fisheries College, Ocean University of China, Qingdao, Shandong, China.

Fisheries College, Ocean University of China, Qingdao, Shandong, China.

出版信息

Comp Biochem Physiol Part D Genomics Proteomics. 2019 Jun;30:14-24. doi: 10.1016/j.cbd.2018.12.003. Epub 2018 Dec 11.

DOI:10.1016/j.cbd.2018.12.003
PMID:30771561
Abstract

Low salinity treatment is proven to be the practical polyploidy inducing method for shellfish with advantages of lower cost, higher operability and reliable food security. However, little is known about the possible molecular mechanism of hypotonic induction. In this study, isobaric tags for relative and absolute quantitation (iTRAQ) based proteomic profiling was pursued to investigate the responses of zygotes of the Pacific oyster Crassostrea gigas to low salinity. A total of 2235 proteins were identified and 87 proteins were considered differentially expressed, of which 14 were up-regulated and 69 were down-regulated. Numerous functional proteins including ADP ribosylation factor 2, DNA repair protein Rad50, splicing factor 3B, tubulin-specific Chaperone D were significantly changed in abundance, and were involved in various biology processes including energy generation, vesicle trafficking, DNA/RNA/protein metabolism and cytoskeleton modification, indicating the prominent modulation of cell division and embryonic development. Parallel reaction monitoring (PRM) analyses were carried out for validation of the expression levels of differentially expressed proteins (DEPs), which indicated high reliability of the proteomic results. Our study not only demonstrated the proteomic alterations in oyster zygotes under low salinity, but also provided, in part, clues to the relatively lower hatching rate and higher mortality of induced larvae. Above all, this study presents a valuable foundation for further studies on mechanisms of hypotonic induction.

摘要

低盐处理已被证明是贝类多倍体诱导的实用方法,具有成本低、可操作性高和食品安全可靠的优点。然而,低盐诱导的可能分子机制知之甚少。在这项研究中,采用基于等重标记相对和绝对定量(iTRAQ)的蛋白质组学分析方法,研究了太平洋牡蛎(Crassostrea gigas)受精卵对低盐的反应。共鉴定了 2235 种蛋白质,其中 87 种被认为差异表达,其中 14 种上调,69 种下调。大量功能蛋白,包括 ADP 核糖基化因子 2、DNA 修复蛋白 Rad50、剪接因子 3B、微管特异性伴侣 D 等,其丰度显著变化,涉及包括能量生成、囊泡运输、DNA/RNA/蛋白质代谢和细胞骨架修饰在内的各种生物学过程,表明细胞分裂和胚胎发育的明显调控。并行反应监测(PRM)分析用于验证差异表达蛋白(DEPs)的表达水平,表明蛋白质组学结果具有较高的可靠性。本研究不仅证明了低盐胁迫下牡蛎受精卵的蛋白质组学变化,而且部分揭示了诱导幼虫孵化率相对较低和死亡率较高的原因。总之,本研究为低盐诱导机制的进一步研究提供了有价值的基础。

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