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Identification of maturation and protein synthesis related proteins from porcine oocytes during in vitro maturation

机译:鉴定猪卵母细胞体外成熟过程中成熟和蛋白质合成相关蛋白

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Background In vitro maturation (IVM) of mammalian oocytes is divided into the GV (germinal vesicle stage), MI (metaphase I stage) and MII (metaphase II stage) stages, and only fully mature oocytes have acquired the ability to be fertilized and initiate zygotic development. These observations have been mostly based on morphological evaluations, but the molecular events governing these processes are not fully understood. The aim of the present study was to better understand the processes involved in the molecular regulation of IVM using 2-DE analysis followed by mass spectrometry to identify proteins that are differentially expressed during oocyte IVM. Result A total of 16 up-regulated and 12 down-regulated proteins were identified. To investigate the IVM process, we specifically focused on the proteins that were up-regulated during the MII stage when compared with the GV stage, which included PRDX 2, GST, SPSY, myomegalin, PED4D, PRKAB 1, and DTNA. These up-regulated proteins were functionally involved in redox regulation and the cAMP-dependent pathway, which are essential for the intracellular signaling involved in oocyte maturation. Interestingly, the PDE4D and its partner, myomegalin, during the MII stage was consistently confirmed up-regulation by western blot analyses. Conclusion These results could be used to better understand some aspects of the molecular mechanisms underlying porcine oocyte maturation. This study identified some regulatory proteins that may have important roles in the molecular events involved in porcine oocyte maturation, particularly with respect to the regulation of oocyte meiotic resumption, MII arrest and oocyte activation. In addition, this study may have beneficial applications not only to basic science with respect to the improvement of oocyte culture conditions but also to mammalian reproductive biotechnology with potential implications.
机译:背景技术哺乳动物卵母细胞的体外成熟(IVM)分为GV(生殖小泡阶段),MI(中期I期)和MII(中期II期)阶段,只有完全成熟的卵母细胞才具有受精和启动的能力。合子发育。这些观察主要基于形态学评估,但对这些过程的分子事件尚不完全了解。本研究的目的是使用2-DE分析和质谱法更好地了解IVM分子调控中涉及的过程,以鉴定在卵母细胞IVM中差异表达的蛋白质。结果共鉴定出16种上调蛋白和12种下调蛋白。为了研究IVM过程,我们特别关注与GV阶段相比在MII阶段上调的蛋白质,包括PRDX 2,GST,SPSY,肌巨蛋白,PED4D,PRKAB 1和DTNA。这些上调的蛋白在功能上参与了氧化还原调节和cAMP依赖性途径,这对于参与卵母细胞成熟的细胞内信号传导至关重要。有趣的是,蛋白印迹分析一直一致地证实MII阶段的PDE4D及其伴侣肌巨蛋白。结论这些结果可用于更好地了解猪卵母细胞成熟的分子机制的某些方面。这项研究确定了一些调节蛋白,这些蛋白可能在与猪卵母细胞成熟有关的分子事件中起重要作用,特别是在卵母细胞减数分裂恢复,MII阻滞和卵母细胞活化方面。此外,这项研究不仅对改善卵母细胞培养条件的基础科学具有有益的应用,而且对具有潜在影响的哺乳动物生殖生物技术也有有益的应用。

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