Expression Of Mice Ovum Mutant Gene In Early Embryonic Development

In order to study the mutant gene (ovum mutant,Om) of DDK females with other inbred strains showed mating male sterility phenomenon of polarity, that is, (DDK♀x alien♂) F1-generation in the developing embryo to pregnancy 3-5 day obstacles due to the death of the blastocyst formation process. In this study, carried out by lethal hybrid embryos (DDK♀x BALB/C♂) as the main study, while the use of normal embryos of the cross (DDK♀x DDK♂) for control of the research work. Through the normal pre-implantation zygotes in mating collection observation, found that the death of hybrid embryos from the 8 cell stage also after the beginning of morula apoptosis occurred; through semi-quantitative RT-PCR technology to compare expression of candidate genes in different combinations of mating pre-implantation fertilized egg.This study is divided into two parts:Test-ⅠCombination of different mating mice pre-implanted fertilized egg collection and observation1. Experiment with the establishment of mating combinations.Om gene in mice resulted embryonic death phenomenon is the strict combination of mating, (DDK♀x BALB/c♂) can be embryonic lethal gene that is the role of Om, and DDK strains obtained from the mating of the F1 cross to the normal development of mouse embryos, without any abnormal development of the situation. Therefore, the choice of the cross as a control, the pre-fertilized egg implants and the development of candidate gene expression in normal embryos can be used as reference standard.2. Pre-implantation of the fertilized eggs during the collection and observationSelect the 60-day-old mice, super-exclusive deal with the normal female female ratio of 1:1, respectively, in accordance with the way male and female mating cage together. According to embryonic development time, microscope observation, collecting eggs at different stages. The results show that DDK ultra-exclusive deal with a large number of female embryos at the 2-cell developmental stagnation and can not conduct a follow-up test. Mating the normal mode hybrid embryos died after 8-cell apoptosis, identified as dead cells. Acquisition and development morphology were normal on 2 cells to 8-cell embryos of various developmental stages 80-100 pieces.Test-ⅡSemi-quantitative RT-PCR detection expression of candidate genes in the different mating combinations in mice pre-implanted fertilized eggs.1. The choice of candidate genesCurrently in the process of early embryonic development in mice, the main function of gene expression of proto-oncogenes, growth factors, growth and apoptosis genes, such as leukocyte inhibitory factor categories, including various functional changes in gene expression caused by different effects. Therefore, the test to select differentially expressed due to their non-normal development of embryos and embryonic Om gene triggered by the phenomenon of death in the embryology and physiology is very similar to the eight functional genes as a candidate gene, hybrid-type embryo death settlement with the normal development of embryos in the process of of candidate differentially expressed gene fragment can be considered as a direct result of fetal death.2. Semi-quantitative RT-PCR detectionThrough semi-quantitative PCR detection comparison in eight candidate genes with the hybrid embryos to death to pay the normal 2-cell embryos,4 cell,8 cell stage fertilized eggs in the relative content of mRNA. The results showed that the test carried by the DDK mouse gene Om hybrid fertilized preimplantation embryos died during development of observation and the impact of a series of pre-implantation embryos in the normal development of the functional differences in gene expression studies, analysis of gene Om triggered by the process of embryonic death, Om revealed a class of genes is affected embryonic development and, indirectly, the functional genes in mouse embryos resulted in the phenomenon of death. To further clarify the role of egg mutation mechanism, and improve the process of mouse blastocyst formation of molecular control theory to lay a certain foundation.