鸡与鹌鹑属间杂交早期胚胎bcl-2、P53基因表达的差异及发育性变化

 【目的】探讨细胞凋亡因子bcl-2、P53对鸡与鹌鹑属间杂交早期胚胎发育的影响。【方法】人工授精获得鸡（♂）与鹌鹑（♀）杂交种蛋，按照鸡标准孵化条件同批入孵，随机采集66、72、78、84、90、96、102、108、114、120 h活胚。采用RT-PCR方法，用Wpkci和β-actin引物进行多重PCR鉴定胚胎样本性别，后选取各时间点雌、雄胚胎各4枚，以β-actin为内标，测定bcl-2、P53的mRNA相对丰度。【结果】（1）雄性胚胎66～114 h bcl-2 mRNA表达维持在较低水平，96 h 有所降低，随后回升到原水平，120 h 极显著升高（P＜0.01），达到峰值；雌性胚胎66～114 h bcl-2 mRNA表达维持在较低水平，114 h 极显著升高（P＜0.01），此后维持在较高水平，120 h 达到峰值。不同日龄之间的比较，114 h雌性表达显著高于雄性（P＜0.05）；（2）雌、雄胚胎P53 mRNA发育性表达模式基本一致。66 h表达水平较高，72 h表达降低，达到低谷，雄性达到显著（P＜0.05），雌性整体差异不显著，随后上升维持在原水平；不同日龄之间表达差异不显著。【结论】114 h雌性胚胎bcl-2 mRNA表达高峰点与其P53 bcl-2表达最低水平相吻合；72 h雌、雄胚胎 P53 mRNA表达与P53 bcl-2表达水平一致，处于低谷点；72、114 h早期杂交胚胎bcl-2、P53基因表达出现异常。

The Expresstional Differences and Developmental Changes of bcl-2,P53 Gene in Early Embryos of Generic Hybrids of Chicken-Quail

Abstract：【OBJECTIVE】To investigate the influences of apoptotic factors bcl-2、P53 on early embryos of generic hybrids;【METHOD】we acquired the cross bred eggs of chicken(♂)-quail(♀)with artificial insemination,hatched the eggs in one batch according to the standard condition of chicken,collected the early living-embryos of 2.75,3.0,3.25,3.50,3.75,4.0,4.25,4.5,4.75 and 5.0 d at random.Adopted the method of RT- PCR,used multi-ply PCR to identify the embryo sex with primers of Wpkci and β-actin,then selected four embryos of female and male respectively of each period,taken β-actin for internal standard to determine the relative quantity of bcl-2、P53 mRNA of embryos;【RESULTS】1)The bcl-2 mRNA expression of 2.75～4.75 d of male embryos maintained lower level,declined on 4.0 d,then reached the initial level,rose distinctly on 5.0 d(P<0.01)and reached the peak;the bcl-2 mRNA expression of 2.75～4.5 d of male embryos maintained lower level,rose distinctly on 4.75 d(P<0.01),then maintained higher level and reached the peak on 5.0 d;the female bcl-2 mRNA expression of 4.75 d were higher distinctly than male of the same days(P<0.05); 2)The patterns of P53 mRNA expression of male and female embryos were basically consistent.They were higher on 2.75 d,declined on 3.0 d(P<0.05)and reached the lowest level,declined distinctly in male,but the whole differences weren’t significant in female,then reached and maintained the initial level.Comparing the expression between different sex, the differences weren't significant.【CONCLUSION】We found two abnormal points in gene expresstional periods in female(3.0 and 4.75 d),but one point in male(3.0 d).The disorder regulated by key factors of apoptosis may induce embryo death.This conclusion should be confirmed further after the research of bcl-2、P53 expression of chicken and quail's embryos of the same period.