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| Screening and Quantitative Analysis of Differential Expression Genes in In vitro Fertilized Bovine Pre-implantation Embryos |
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Abstract The large capacity of differential expression genes during oocyte maturation and the transition from maternal to zigotic control makes it possible to screen genes playing crucial roles in embryonic development by mRNA differential display reverse transcription PCR(DDRT-PCR). In order to explore the gene expression mechanism of in vitro maturation (IVM) of oocytes and in vitro development of pre-implantation embryos derived from in vitro fertilization (IVF) in different developmental stages in cattle, mRNA DDRT-PCR was employed to screen differential expression genes in germinal vesicle (GV) stage oocytes, 8-cell stage embryos and blastocysts. Real-time quantitative PCR (Q-PCR) was used to analyze the expression abundance of the differential expression genes in the three stages and in vitro matured MⅡ oocytes. Five differential displayed straps were discovered by DDRT-PCR. The results of sequencing and database analysis showed that the 5 fragments were corresponding to pyrophosphatase (inorganic) 1(PPA1), erbb2 interacting protein (ERBB2IP), restin-like (CEP350), ribosomal protein L27a (RPL27A) and IK cytokine, down-regulator of HLAⅡ (IK), respectively. Q-PCR and SPSS statistical analysis indicated that the mRNA level of PPA1, ERBB2IP and CEP350 decreased along with the procedure of embryonic development, whereas that of RPL27A turned to increase till blastocyst stage. The mRNA level of IK fluctuated before blastocyst stage and then decreased. Overall, the results of this research provide more information on effects of oocyte maturation on the following embryonic development and the mechanism of zygotic gene activation.
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Received: 02 September 2011
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