为了明确ghrelin是否参与了卵母细胞成熟及胚胎早期发育进程,本研究利用免疫荧光技术和实时定量RT-PCR技术检测了绵羊卵母细胞和体内早期胚胎中ghrelin蛋白的表达定位和ghrelin mRNA水平相对表达变化规律。免疫荧光染色结果表明,ghrelin蛋白主要分布于卵母细胞胞质内;实时定量RT-PCR结果揭示绵羊卵母细胞和早期胚胎ghrelin mRNA的相对表达量依据发育阶段的不同而呈现一定变化规律,即在成熟卵母细胞,2细胞胚胎期和8细胞胚胎期显著高于未成熟卵母细胞和4细胞胚胎期(P〈0.05),囊胚期表达量最高。卵母细胞和早期胚胎中ghrelin蛋白的表达及ghrelin mRNA特定的表达模式,揭示这一新型分子在绵羊卵母细胞成熟以及胚胎早期发育过程中具有潜在的调控作用。
The possibility of ghrelin expressed in sheep oocytes and pre-implantation embryos produced in vivo was investigated in this study. The observed ghrelin immunoreactivity of oocytes and embryos at all stages was predominantly in the cytoplasm. Relative real-time reverse-transciptase (RT)-PCR experiments confirmed that the ghrelin mRNA levels varied depending on the developmental stage, with the highest expression in the blastocyst, metaphase Ⅱ (MⅡ) oocytes, 2- and 8-cell stages had a significantly higher expression in the germinal vesicle (GV) oocytes, 4-stage and morula. Dynamic changes and the persistent presence of the ghrelin signaling system within oocytes and pre-implantation embryos opens up the possibility of a potential regulatory role of this novel molecule during oocyte maturation and embryonic development.