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斑马鱼早期发育过程中Pou5f1蛋白的表达及翻译后修饰

Pou5f1 protein expression and posttranslational modification during early zebrafish development.

作者信息

Lippok Bernadette, Song Sungmin, Driever Wolfgang

机构信息

Developmental Biology, Institute Biology I, Faculty of Biology, Albert-Ludwigs-University Freiburg, Freiburg, Germany.

出版信息

Dev Dyn. 2014 Mar;243(3):468-77. doi: 10.1002/dvdy.24079. Epub 2013 Nov 21.

Abstract

BACKGROUND

Pou5f1/Oct4 is a transcription factor essential for maintenance of pluripotency in mammals and for control of blastula and gastrula stage gene regulatory networks in zebrafish. Information on Pou5f1 protein distribution was before this study not available for zebrafish. Therefore, we generated polyclonal antibodies that selectively recognize Pou5f1 and analyzed its protein distribution and modification during development.

RESULTS

Pou5f1 protein is present in unfertilized oocytes, and persists in all embryonic and enveloping layer cell nuclei until the end of gastrulation, but is absent from yolk syncytial nuclei. Pou5f1 is subject to multiple developmentally regulated phosphorylations, with the higher phosphorylated forms prevailing in the oocyte and during late gastrulation.

CONCLUSIONS

The developmental protein profile correlates with the stages during which deep cells are not committed to a specific germ layer. The posttranslational modification by phosphorylation opens the possibility that Pou5f1 may be subject to temporal or region specific modulation of its activity or stability by embryonic signaling mechanisms.

摘要

背景

Pou5f1/Oct4是一种转录因子,对于维持哺乳动物的多能性以及控制斑马鱼囊胚和原肠胚阶段的基因调控网络至关重要。在本研究之前,尚无关于斑马鱼Pou5f1蛋白分布的信息。因此,我们制备了能选择性识别Pou5f1的多克隆抗体,并分析了其在发育过程中的蛋白分布及修饰情况。

结果

Pou5f1蛋白存在于未受精的卵母细胞中,并在所有胚胎细胞核和包被层细胞核中持续存在,直至原肠胚形成结束,但卵黄多核体细胞核中不存在该蛋白。Pou5f1会经历多种发育调控的磷酸化过程,在卵母细胞和原肠胚后期,磷酸化程度较高的形式占主导。

结论

发育过程中的蛋白谱与深层细胞尚未确定为特定胚层的阶段相关。磷酸化的翻译后修饰表明,Pou5f1的活性或稳定性可能会通过胚胎信号机制受到时间或区域特异性的调控。

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