雌激素受体在鸡与鹌鹑属间杂交早期胚胎中的差异表达及其作用

Effect and Different Expression of ER Gene in Early Embryos of Generic Hybrids of Chicken-quail

旨在研究雌激素受体（ER）与鸡-鹌鹑属间杂交早期胚胎发育、性别分化的关系。人工授精获得鸡（♂）-鹌鹑（♀）杂交种蛋，按照鸡的孵化标准同批入孵，连续采集早期活胚（2．75、3．00、3．25、3．50、3．75、4．00、4．25、4．50、4．75、5．00d），采用RT—PCR法，用Wpkci和β-actin引物进行多重PCR，鉴定早期胚胎性别；之后选取各时间点雌、雄胚胎各4枚，以β-actin为内标，测定ER mRNA相对表达丰度。雌、雄胚胎ER mRNA的发育性变化趋势基本一致，2．75～3．00d表达水平均较高；3．25d雄性表达水平显著降低（P〈0．05），雌性则极显著降低（P〈0．01）；3．50d雌、雄表达水平又回复至较高水平；3．75d二者均极显著降低（P〈0．01），此后一直维持在较低水平。不同日龄间比较，2．75～3．50d雌性ER表达高于雄性，差异极显著（P〈0．01）。杂交胚胎性分化时间大致开始于胚胎发育的第2．75～3．50d，性分化期ER的异常表达可能与胚胎死亡、杂种不育有关，这一结论有待于进一步研究发育同期鸡、鹌鹑胚胎ER的表达及其功能后证实。

To investigate the effect of estrogen receptor（ER） on early embryos of generic hybrids of chicken-quail, we acquired cross bred eggs of chicken（ ♂ ）-quail（ ♀ ） by artificial insemination, hatched the eggs in one batch according to the standard condition of chicken, collected early livingembryos of 2.75,3.00,3.25,3.50,3.75,4.00,4.25,4.50,4. 75 and 5.00 d at random. Adopting RT-PCR and multi-ply PCR, we identified the sex of embryos with primers of Wpkci and β-actin,then selected four female and male embryos respectively from each period taken β-actin as internal standard to determined the relative quantity of ER mRNA. The patterns of ER mRNA expression of male and female were basically consistent. The expressive level were all higher in 2.75 ～3.00 d,declined distinctly in male（P〈0.05） and very significantly in female（P〈0.01） in 3.25 d,then ,all reached initial level in 3.50 d and declined great significantly in 3.75 d （P〈0.01）, then maintained the lower level. Compared the expression between different sex of the same periods, ER mRNA expression of female were higher than male in 2.75～3.50 d ,and the difference was very significant （P〈0.01）. We found 2.75～3.50 d were the gonadal differentiation period. The abnormal ER expression may induced embryos death and hybrid infertility in the periods of gonadal differentiation. Those conclusion should be confirmed after further research on ER ex pression and function in chicken and quail＇s embryos respectively of the same period.