鹌鹑早期胚胎生成过程中性别分化相关基因的表达分析

摘要

In this study the expression profiles of 3β-HSD, P-450cl7, P450arom and Estrogen receptor (ER) genes were analyzed to understand the potential influence of 3β-HSD, P-450c17, P450arom and Estrogen receptor (ER) on the early embryonic development and sex differentiation in quail embryo. The sexes of quail embryos were identified by PCR amplification of Wpkci gene, then 10 female and 10 male embryos at every time point (day 3, 4, 5, 6, 7 and 8 of incubation) of the early development stage were sampled. Real-time PCR was applied to analyze the expression profiles of 3β-HSD、P-450c17、P450arom and Estrogen receptor (ER) genes at different time points of the early development stage (day 3-8 of incubation) in quail embryos. Results showed that the expression levels of P-450c17, P450arom and ER were all higher in female than that in male on day 4 of incubation: for P450arom gene, it was detected only in female embryos throughout the incubation period and displayed the peaking level on day 4 of incubation (P< 0. 01) ; for ER gene, the highest expression levels were detected on day 4 of incubation both in female and male, but the expression level in female was significantly higher than that in male (P< 0.05); for P-450c17, the expression level was higher in female than in male, but the difference was not significant(P>0. 05). The expression level of 3β-HSD gene was significantly higher in female than in male on day 5 of incubation(P<0. 01). These results indicate that 3β-HSD, P-450cl7, P450arom and ER genes play an important role in the process of sex differentiation during the early development stage of quail embryo, especially at the day 4 to day 5 of incubation.%旨在对影响鹌鹑早期胚胎发育及性别分化的4个基因的表达进行研究.采用Wpkci基因和多重PCR鉴定鹌鹑早期胚胎的性别,在此基础上,选取早期胚胎发育不同时间点(3、4、5、6、7、8 d)雌雄鹌鹑胚胎共120枚,其中雌雄各60枚,每个时间点10枚.采用Real-time PCR方法检测鹌鹑雌雄胚胎发育过程中不同时间点性别分化相关基因(3β-HSD、P-450c17、P450arom和ER)的表达,探讨了3β-HSD、P-450c17、P450arom和ER基因对鹌鹑早期胚胎发育及性别分化的影响.结果表明,鹌鹑胚胎P-450c17、P450arom和ER基因在鹌鹑胚胎发育的第4天雌性均高于雄性,其中P450arom基因仅在雌性胚胎中检测出有表达,且表达量在第4天达到最高峰(P＜0.01),雄性胚胎整个发育过程中都无P450arom基因的表达；ER基因在鹌鹑胚胎发育第4天雌性显著高于雄性(P＜0.05)；P-450c17在鹌鹑胚胎发育第4天虽雌性高于雄性,但无显著性差异(P＞0.05).另外,3β-HSD在鹌鹑胚胎发育第5天达到其表达最高值,且雌性极显著高于雄性(P＜0.01).研究结果表明,3β-HSD、P-450c17、P450arom和ER基因在鹌鹑早期胚胎性别分化过程中起着重要的作用,尤其是在胚胎发育4～5 d.