Oct 4在牦牛卵母细胞及早期胚胎发育过程中的表达  

The Expression of Octamer-binding Transcription Factor 4 Gene(Oct 4)in Yak(Bos grunneins)Oocytes and Preimplantation Embryos Produced by in vitro Fertilization

作　　者：赵天[1] 李谷月 潘阳阳[1] 李月琴[1] 陈平[1] 温泽星[1] 崔燕[1] 余四九[1] 

机构地区：[1]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,兰州730070

出　　处：《畜牧兽医学报》2017年第3期577-584,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31272616;31472244)

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。The aim of this study was to explore the effect of octamer-binding transcription factor 4（Oct 4）during yak oocytes maturation and embryos development,which was achieved by examined the expression of transcription factors Oct 4at different stages.In this study,yak cumulusoocyte complexes（COCs）were collected and matured in vitro,the embryos were produced by the method of in vitro fertilization（IVF）and the development competence of oocytes and embryos at different stages were evaluated.The expression and distribution of Oct 4in the level of mRNA and protein in COCs and embryos at different stages were detected by quantitative real-time PCR（qRT-PCR）and indirect immunofluorescence.The results showed as follows：1）The rates of embryos developed to stages of 2-4-cells,5-8-cells,9-16-cells and blastocyst were（64.66±1.76）%,（43.33±1.76）%,（24.66±2.40）%,（9.33±0.67）%,respectively.2）The expression of Oct 4gene could be detected at all stages during yak oocytes maturation and embryos de-velopment,which was highest in 9-16-cells embryos,followed by blastocyst,2-4-cells embryos and 5-8-cells embryos,while it were lowest in immature COCs and mature COCs.3）The result of indirect immunofluorescence method showed that the protein of Oct 4was mainly distributed in the cytoplasm of COCs and embryos,which was weaker in the nucleus.Analyzing by immunofluorescence IOD（integrated optical density）showed that the IOD of 9-16-cells embryos was highest,followed by blastocyst,2-4-cells embryos,which was lowest in 5-8-cells embryos.The results showed that Oct 4could express in various periods of yak oocytes and embryo development,while the expression levels of Oct 4were different,which indicated that Oct 4might participate in the embryonic genetic transcription activation at stages of 9-16-cells,which might take part in maintaining tipotency of inner cell mass in blastocyst.

关 键 词：OCT 4 牦牛 体外受精 胚胎发育 

分 类 号：S823.852[农业科学—畜牧学]