Leptin对陕北白绒山羊体细胞核移植的影响

Influence of Leptin to Somatic Cell Nuclear Transfer in Shanbei White Cashmere Goat

Leptin的功能较为复杂,主要调控机体能量代谢。有研究表明Leptin在卵母细胞成熟及胚胎发育过程中也具有重要作用。本研究在卵母细胞体外成熟基础培养液中添加了不同浓度的Leptin,其中未添加Leptin的设为Ⅰ组,添加10ng/mL和50ng/mL Leptin的分别设为Ⅱ组和Ⅲ组。以陕北白绒山羊皮肤成纤维细胞为供体细胞,三组体外培养成熟的卵母细胞作为受体,利用显微操作方法对成熟卵母细胞进行去核操作,然后将供体细胞注射到卵周隙内,经电融合后形成体细胞克隆胚。根据卵母细胞体外成熟率、核移植效率以及克隆胚囊胚发育率分析Leptin在山羊核移植中的作用。结果表明Ⅰ组山羊卵母细胞体外成熟率和核移植效率显著高于其他两组(P<0.05),三组克隆囊胚发育率无显著差异(P>0.05)。Leptin降低了山羊卵母细胞体外成熟和核移植效率,对克隆胚发育能力无影响。

The roles of Leptin are very complex. One of the important roles is regulation of energy metabolism. Some in- vestigations showed that Leptin also had crucial role during the oocyte maturation and embryo development. Basic oocyte maturation medium with different concentrations of Leptin （0, group I ; 10 ng/mL, group II and 50 ng/mL, group III ） were used to mature Shanbei white cashmere goat oocytes. Skin fibroblasts from Shanbei white cashmere goat were used as donor cells, and enucleated matured oocytes from three groups were used as recipient ooplasm. Donor cells were introduced into perivitelline space of recipient ooplasm by microinjeetion, and the somatic cell cloned embryos were constructed after e- lectric fusion. Subsequently, the oocyte maturation rate in vitro, efficiency of somatic cell nuclear transfer and blastocyst rate of cloned embryos were analyzed to investigate the role of Leptin in nuclear transfer of goat. The results showed that the percentage of oocyte maturation in vitro and efficiency of somatic cell nuclear transfer were significantly higher in group I than the other two groups （P 〈 0.05 ）, and the percentage of blastocyst were no statistic different among the three groups （P 〉 0. 05 ）. These results indicated that Leptin reduced maturation rate of goat oocyte in vitro and the efficiency of somatic cell nuclear transfer. However, Leptin had no effect on the developmental ability of cloned embryo.