关键词: 猪, 孤雌激活胚, 组蛋白乙酰化, 实时定量PCR, 组蛋白去乙酰化酶(HDAC1), mRNA表达

Abstract:

【Objective】 The aims of this study were to investigate the relationship between the levels of histone acetylation (AcH4K5) and mRNA expression level of histone deacetylase 1(HDAC1) during early development of porcine parthenogenetic(PA) embryos in vitro. 【Method】 Porcine PA embryos (2-cell, 4-cell, morula and blastocyst) were collected, histone acetylation levels of AcH4K5 of four states’ embryos were assessed using immunofluorescence and laser confocal microscope by detecting the fluorescence intensity. Real-time PCR method was used for quantitative analysis of HDAC1 mRNA level in PA embryos. 【Result】The results showed that AcH4K5 expressed in nucleolus of all embryos from 2-cell to blastocyst, and the levels of AcH4K5 expression increased gradually, reached the highest in blastocyst stage, and the expression at each stage was different significantly(1124.77±127.78, 1305.81±184.23, 1795.19±318.67, and 2149.63±529.47, P＜0.05, respectively). There were dynamic changes of HDAC1 mRNA level among four states’embryos (2-cell, 4-cell, morula and blastocyst of PA embryos), and mRNA expression patterns of HDAC1 genes decreased from 2-cell to blastocyst stage (1.00±0, 0.91±0.009, 0.85±0.008, 0.67±0.006, P＜0.05, respectively). 【Conclusion】 These results indicated that there is an inverse correlation between the levels of AcH4K5 and mRNA expression of HDAC1 genes from 2-cell to blastocyst.

Key words: porcine, parthenogenetic embryos, histone acetylation, real-time fluorescence quantitative PCR, histone deacetylase 1 (HDAC1), mRNA expression