不同浓度Leptin对猪卵母细胞体外成熟和早期孤雌发育的影响

Effects of Leptin Supplementation in in vitro Maturation Medium on Meiotic Maturation of Oocytes and Preimplantation Development of Parthenogenetic Embryos in Pigs

[目的]系统比较了在成熟培养液中添加Leptin对猪卵母细胞体外成熟和孤雌激活胚的发育率和囊胚质量,同时比较了成熟培养液和胚胎培养液添加Leptin对孤雌激活胚发育的影响。[方法]成熟培养液中添加不同浓度Leptin对猪卵母细胞体外成熟和早期孤雌发育能力的影响及囊胚的内细胞团细胞数目,并且研究了成熟培养液和胚胎培养液单独或联合添加Leptin对猪早期孤雌胚胎发育能力的影响及囊胚的内细胞团细胞数目。[结果]成熟培养液中添加不同浓度Leptin(0,10,50,100和200 ng/ml),体外成熟44 h,卵母细胞核成熟率统计分析差异不显著(P>0.05);将核成熟卵母细胞进行化学激活,第2天胚胎卵裂率统计分析差异不显著(P>0.05),但100ng/ml组与其他组第7天的囊胚率及囊胚的总细胞数差异显著(P<0.05);成熟液添加Leptin 100 ng/ml、胚胎培养液添加Leptin 10 ng/ml与其他组第7天的囊胚率及囊胚的内细胞团细胞数差异显著(P<0.05)。[结论]Leptin对卵母细胞成熟和孤雌胚胎发育能力上有相辅相成的作用。

[ Objective] The study was designed to examine the effects of leptin addition in in vitro maturation medium on nuclear maturation of porcine oocytes and to evaluate the quality of porcine parthenogenetic activation （PA） blastocysts. In addition, the effects of leptin addition in both IVM and in embryo culture medium on the developmental potential on the porcine PA embryos were investigated. [Method] The effects of leptin addition in in vitro maturation medium on nuclear maturation of porcine oocytes,preimplantation development of parthenogenesis and the total cell number of each blastocyst were studied. And the effects of leptin addition in IVM medium or in embryo culture medium alone or in the meanwhile on preimplantation development of parthenogenesis and the total cell number of each blastocyst were investigated. [Result] Porcine oocytes were matured in IVM medium contained different concentrations of leptin（0,10,50,100 and 2130 ng/ml） for 44h. The rate of nuclear mature in various groups showed no significant difference （ P 〉 0.05）. Oocytes with the first polar body were parthenogenetically activated in vitro by 5 μmol/L ionomycin for 5 min and 2 mmol/L 6-dimethylaminopurine （DMAP） for 4 h.The rate of cleavage on day 2 in different groups showed no significant difference （ P 〉0.05）.Furthermore,the rate of blastocyst fonuation on day 7 was significantly higher in the leptin 100 ng/ml group than that in the other groups （ P 〈0.05）.The total cell number of each blastocyst matured in the leptin 100 ng/ml group and leptin 10 ng/ml/n vitro embryo culture in the same time, and other groups on day 7 showed a significant difference （P 〈 0.05）. [Conclnsion]Leptin had a synergistic role on both oocyte maturation and preimplantation embryo development.