CAJ | 학술논문

为了更全面的对四倍体胚胎的发育能力进行评估,本研究利用原位末端标记(TUNEL)法检测了电融合生产的小鼠四倍体囊胚细胞凋亡情况,同时用实时定量PCR方法检测了促凋亡基因Bax和抑凋亡基因Bcl-2在四倍体囊胚中的表达.结果显示:四倍体与体外二倍体胚胎发育到囊胚的比率没有显著差别(P>0.05);四倍体囊胚无论是内细胞团细胞数、滋养层细胞数还是内细胞团/总细胞数比率都显著低于体内与体外二倍体囊胚(P<0.05);四倍体囊胚细胞凋亡指数显著高于体内与体外二倍体囊胚细胞凋亡指数(P<0.05);促凋亡基因Bax在四倍体囊胚中的表达显著高于体内与体外二倍体囊胚中的表达(P<0.05),而抑凋亡基因Bcl-2在四倍体囊胚中的表达同体内与体外二倍体囊胚中的表达没有显著差异(P>0.05).因此,四倍体囊胚质量比体内与体外二倍体囊胚质量都差,这可能是四倍体胚胎不能发育到足月的原因之一.
위료경전면적대사배체배태적발육능력진행평고,본연구이용원위말단표기(TUNEL)법검측료전융합생산적소서사배체낭배세포조망정황,동시용실시정량PCR방법검측료촉조망기인Bax화억조망기인Bcl-2재사배체낭배중적표체.결과현시:사배체여체외이배체배태발육도낭배적비솔몰유현저차별(P>0.05);사배체낭배무론시내세포단세포수、자양층세포수환시내세포단/총세포수비솔도현저저우체내여체외이배체낭배(P<0.05);사배체낭배세포조망지수현저고우체내여체외이배체낭배세포조망지수(P<0.05);촉조망기인Bax재사배체낭배중적표체현저고우체내여체외이배체낭배중적표체(P<0.05),이억조망기인Bcl-2재사배체낭배중적표체동체내여체외이배체낭배중적표체몰유현저차이(P>0.05).인차,사배체낭배질량비체내여체외이배체낭배질량도차,저가능시사배체배태불능발육도족월적원인지일.
The purpose of this study was to investigate the developmental potential of the mouse tetraploid embryos by the detection of the cell apoptosis and the expression of Bax and Bcl-2 genes in the tetraploid blastocysts.The results indicated that the rate of the blastocyst formation was no significant difference between in the tetraploid embryos and in the diploid embryos in vitro.The result of the differential staining showed that the cell numbers of ICM and TE,the ratio of ICM/Total cells in the tetraploid blastocysts were lower than that in the diploid blastocysts in vitro or in vivo.At the same time,the detection of the cell apoptosis by TUNEL suggested that the index of the cell apoptosis in the tetraploid blastocysts was higher than that in the diploid blastocysts in vitro or in vivo.In addition,the expression of Bax gene in the tetraploid blastocysts was higher than that in the diploid blastocysts in vitro or in vivo,whereas there was no significant different expression of Bcl-2 gene among them.The study demonstrated that the quality of the mouse tetraploid blastocysts was much worse than that of the diploid blastocysts in vitro or in vivo,which may cause the abnormal development of the mouse tetraploid embryos.