Oct 4在牦牛卵母细胞及早期胚胎发育过程中的表达

发布时间：2018-03-30 16:24

本文选题：Oct　切入点：牦牛　出处：《畜牧兽医学报》2017年03期

【摘要】：旨在通过检测转录因子Oct 4(Octamer-binding transcription factor 4)在牦牛(Bos grunneins)卵母细胞成熟及早期胚胎发育过程中的表达,分析Oct 4在卵母细胞与体外受精胚胎发育过程中的作用。本研究收集牦牛卵丘-卵母细胞复合体(Cumulus-oocyte complexes,COCs)对其进行体外成熟培养,生产体外受精(In vitro fertilization,IVF)胚胎,评估各时期胚胎发育能力。采用实时荧光定量PCR(Quantitative real-time PCR,qRT-PCR)和间接免疫荧光染色方法从基因和蛋白水平检测COCs和胚胎各时期Oct 4的表达与分布。结果表明,1)2~4-细胞胚胎、5~8-细胞胚胎、9~16-细胞胚胎和囊胚的发育率分别为(64.66±1.76)%、(43.33±1.76)%、(24.66±2.40)%和(9.33±0.67)%。2)COCs和胚胎各发育时期均可检测到Oct 4基因,其中9~16-细胞胚胎表达最高,依次为囊胚、2~4-细胞胚胎和5~8-细胞胚胎,未成熟COCs和成熟COCs中表达最低。3)免疫荧光染色结果显示,OCT 4蛋白主要分布在COCs和体外受精胚胎的细胞质中,在细胞核中表达较弱。免疫荧光IOD值分析可得,9~16-细胞胚胎IOD值最高,依次为囊胚、2~4-细胞胚胎,5~8-细胞胚胎最低。综上表明,Oct 4在牦牛卵母细胞和早期胚胎发育过程中的各个时期均有表达,且不同时期该基因的表达量存在差异。由此说明Oct 4在9~16-细胞胚胎时期可能参与胚胎合子型基因转录激活的调控过程,在囊胚时期可能参与维持内细胞团全能性的过程。
[Abstract]:The aim of this study was to detect the expression of Oct 4(Octamer-binding transcription factor 4 in the maturation and early embryo development of Bos grunneinsoocytes, and to analyze the role of Oct 4 in the development of oocytes and embryos during in vitro fertilization.In this study, yak cumulus-oocyte complexes (Cumulus-oocyte complex COCs) were collected and cultured in vitro to produce in vitro fertilization (IVF) embryos and to evaluate the development ability of embryos.The expression and distribution of COCs and Oct 4 were detected at gene and protein levels by real-time fluorescence quantitative PCR(Quantitative real-time PCR qRT-PCR and indirect immunofluorescence staining.Immunofluorescence IOD analysis showed that the highest IOD value was found in the 16-cell blastocyst embryos, followed by the lowest IOD values in the blastocyst embryos and the blastocyst 4- cell embryos.The results showed that Oct4 was expressed in yak oocytes and early embryos at all stages, and the expression of Oct4 was different in different stages.It is suggested that Oct _ 4 may be involved in the regulation of zygote type gene transcriptional activation at the embryo stage and in the blastocyst stage during the blastocyst stage.
【作者单位】：甘肃农业大学动物医学院甘肃省牛羊胚胎工程技术研究中心;
【基金】：国家自然科学基金(31272616;31472244)
【分类号】：S823.85

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