摘要
为详细了解玻璃化冷冻对绵羊卵母细胞中母源基因mRNA表达量的影响,分别玻璃化冷冻GV期和IVM期(18、24h)绵羊卵母细胞,解冻后进行体外培养。GV期和IVM期卵母细胞经过冷冻-解冻后,卵裂率(10.37%、23.17%、33.07%)均极显著低于对照组(82.96%,P<0.01),且冷冻-解冻后的GV期卵母细胞卵裂率极显著低于冷冻-解冻后的IVM期卵母细胞(P<0.01)。本试验利用荧光实时定量PCR技术检测4种母源基因:Gdf9(生长分化因子-9)、Zar1(合子阻泄因子)、Mater(胚胎必要的母体抗原)、Dnmt1(DNA甲基化转移酶1)在不同处理后的绵羊卵母细胞中的mRNA含量。结果表明,4个基因的mRNA在GV期的表达量均高于IVM期的卵母细胞(P<0.01);经玻璃化冷冻处理后,4个基因的mRNA表达量升高,其中GV期含量最高(P<0.01)。结果提示,绵羊GV或IVM期卵母细胞玻璃化冷冻导致母源基因mRNA表达量升高,可能会对胚胎发育产生负面影响。
To investigate the effect on mRNA expression of maternal genes in ovine oocytes by vitrified freezing,the ovine oocytes at stage GV and IVM（18,24 h） were disposed with the method of vitrified freezing and cultured in vitro under normal conditions after defrosting,and the result showed that the vitrified oocytes at stage GV and IVM both had lower cleavage rate（10.37%,23.17%,33.07%） than that in control group（82.96%,P0.01）.In addition,vitrified oocytes at stage GV had lower cleavage rate than that at stage IVM（P0.01）.The content of mRNA of four maternal genes,Gdf9（growth differentiation factor-9）,Zar1（zygote arrest factor-1）,Mater（maternal antigen that embryo requires）,Dnmt1（DNA methyltransferase-1） in ovine oocytes treated differently were detected by fluorescent quantitative real-time PCR in this experiment.The results showed that the content of the four genes mRNA increased using vitrification,especially that in oocytes at stage GV was the highest（P0.01）.The result indicate that the bovine oocytes at stage GV or IVM with higher transcripts by vitrification,may have negative effect on embryonic development.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2011年第9期1233-1238,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
新疆自治区自然科学基金(200821182)
新疆自治区科技攻关(200841122)
新疆自治区科技计划项目(200711104)
国家支撑计划(2008BADB2B05-10)
国家"十一五"863计划(2008aa101005)
新疆自治区动物生物技术重点开放实验室开放课题(XJYS0301-2009-01)
关键词
母源基因
玻璃化冷冻
荧光实时定量PCR
maternal genes
vitrified freezing
fluorescent real-time quantitative PCR
