Develomental Research Of Bcl-2,P53,ER In Early Embryos Of Hybrids Of Chicken-quail

1. Adopted gene Wpkci to identify the sex of early embryos from generic hybrids of chicken-quail We researched the sex of early embryos of hybrids with multiply PCR, made known sex quail as external control and takenβ-actin for internal standard, used primers that were designed according to the conservative area of gene Wpkci of quail.Results: 1) A simple,fast,credible and stable method to identify the sex of earlier hybrids embryos had been established by using primer Wpkci, 2) In earlier embryos,the death rate of female was higher than male and there was no change peak.There were more male than female in earlier embryos,the sexual proportion of earlier embryos compared with academic numerical value was significantly different (P<0.01),there was no difference between different periods(P>0.05).2. The expresstional differences and developmental changes of bcl-2 and P53 in early embryos of generic hybrids of chicken-quailbcl-2 and P53 are the key genes of apoptosis, at present there is no reports about the expressive pattern of bcl-2 and P53 of each period of early embryos.We had collected early living-embryos of 66,72,78,84,90,96,102,108,114 and 120 h at random.Adopted the method of RT-PCR, takenβ-actin as internal standard to determined the relative quantity of bcl-2 and P53 mRNA on the foundation of embryos sex.Results: 1)At 114 h the expressional peak of bcl-2 mRNA of female embryos coincide with its P53?bcl-2 lowest expressional level, 2) At 72 h P53 mRNA expressional quantity of male and female embryos were consistent with their P53?bcl-2 expressional level, both were at low level, 3)At 72,114 h the bcl-2, P53 mRNA expression of early embryos of hybrids were abnormal.3. The effect and different expression of ER in early embryos of generic hybrids of chicken-quail The research indicated that estrogen receptor have an eternal effect on avian sex differentiation.Therefore we adopted the method of RT- PCR to study the relative quantity of embryo ER mRNA.Results: We found the gonad differentiation of hybrids was 66～84 h of embryos development. ER expressed abnormally and the female ER mRNA expression were significantly higher than male in the periods of gonad differentiation (P<0.01), then both abnormally declined at 78 h (Male: P<0.05, Female: P<0.01).