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淡水螺卵卵黄前体蛋白组学研究:揭示从水生到陆生产卵的转变。

Egg perivitelline fluid proteome of a freshwater snail: Insight into the transition from aquatic to terrestrial egg deposition.

机构信息

HKBU Institute of Research and Continuing Education, Shenzhen, P. R. China.

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

出版信息

Rapid Commun Mass Spectrom. 2020 Apr 15;34(7):e8605. doi: 10.1002/rcm.8605.

Abstract

RATIONALE

Proteins from the egg perivitelline fluid (PVF) are assumed to play critical roles in embryonic development, but for many groups of animals their identities remain unknown. Identifying egg PVF proteins is a critical step towards understanding their functions including their roles in evolutionary transition in habitats.

METHODS

We applied proteomic and transcriptomic analysis to investigate the PVF proteome of the eggs of Pomacea diffusa, an aerial ovipositing freshwater snail in the family Ampullariidae. The PVF proteins were separated with the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) method, and proteomic analysis was conducted using an LTQ Velos ion trap mass spectrometer coupled with liquid chromatography. Comparison of PVF proteomes and evolution analyses was performed between P. diffusa and other ampullariids.

RESULTS

In total, 32 egg PVF proteins were identified from P. diffusa. They were categorized as PV1-like subunits, immune-responsive proteins, protein degradation, signaling and binding, transcription and translation, metabolism, oxidation-reduction and proteins with unknown function. Interestingly, the proteome includes a calcium-binding protein important in forming the hard eggshell that enabled the terrestrial transition. However, it does not include PV2, a neurotoxic protein that was assumed to be present in all Pomacea species.

CONCLUSIONS

The PVF proteome data from P. diffusa can help us better understand the roles that reproductive proteins played during the transition from underwater to terrestrial egg deposition. Moreover, they could be useful in comparative studies of the terrestrialization in several groups of animals that occurred independently during their evolution.

摘要

原理

卵周质液(PVF)中的蛋白质被认为在胚胎发育中起着关键作用,但对于许多动物群体,其身份仍然未知。鉴定卵周质液蛋白是了解其功能的关键步骤,包括它们在栖息地进化转变中的作用。

方法

我们应用蛋白质组学和转录组学分析方法来研究 Pomacea diffusa(一种 Ampullariidae 科的空中产卵淡水蜗牛)卵的 PVF 蛋白质组。使用十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)方法分离 PVF 蛋白,并使用 LTQ Velos 离子阱质谱仪与液相色谱联用进行蛋白质组分析。比较了 P. diffusa 和其他喇叭蜗牛的 PVF 蛋白质组和进化分析。

结果

总共从 P. diffusa 鉴定出 32 种卵周质液蛋白。它们被归类为 PV1 样亚基、免疫反应蛋白、蛋白降解、信号和结合、转录和翻译、代谢、氧化还原以及功能未知的蛋白。有趣的是,该蛋白质组包括在形成硬蛋壳中起重要作用的钙结合蛋白,这使蜗牛能够进行陆地过渡。然而,它不包括假定存在于所有 Pomacea 物种中的神经毒性蛋白 PV2。

结论

P. diffusa 的 PVF 蛋白质组数据可以帮助我们更好地理解生殖蛋白在从水下到陆地产卵的过渡过程中所起的作用。此外,它们在几个独立进化过程中发生的动物陆地化的比较研究中可能很有用。

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