猪早期孤雌胚胎脂滴含量和脂肪酸代谢相关基因表达的初步研究  

Preliminary Study on Lipid Droplets Content and Fatty Acid Metabolism Related Gene Expression in Porcine Parthenogenetic Embryos

作　　者：刘芳 邢颖 相金柱 李雪玲 LIU Fang;XING Ying;XIANG Jin-zhu;LI Xue-ling(Inner Mongolia University State Key Laboratory for Reproductive Regulation and Breeding of Grassland Livestock,Hohhot 010070,China)

机构地区：[1]内蒙古大学省部共建草原家畜生殖调控与繁育国家重点实验室

出　　处：《内蒙古大学学报（自然科学版）》2019年第4期387-396,共10页Journal of Inner Mongolia University：Natural Science Edition

基　　金：国家自然科学基金(No.81471001);动植物转基因材料价值评估项目(2016ZX08010005)资助项目

摘　　要：作为能量来源,脂质在卵母细胞成熟、胚胎发育和干细胞增殖中发挥着重要作用,猪卵母细胞和早期胚胎中存在丰富的脂质,但是其脂肪酸代谢的作用机制还知之甚少.本研究通过统计卵母细胞的成熟率和体外受精的囊胚率,确定添加10%卵泡液是最优的卵母细胞成熟体系.通过检测电激活、电激活结合CB和9%乙醇分别作用3min、8min、11min等对卵裂率和囊胚率的影响,确定电激活(卵裂率为81.69%±0.41%,囊胚率为26.45%±0.28%)方式是最优的孤雌激活方案.免疫荧光染色显示孤雌囊胚的细胞核数目明显少于体外受精囊胚,对孤雌胚胎进行脂滴染色和脂滴含量检测的结果显示,脂滴定位于核周围,H2Av只存在于细胞核内,不存在于脂滴中,8-细胞时期脂滴直径达到最大,而桑椹胚中脂滴含量最高.对脂肪酸代谢相关基因的RT-PCR结果显示,与卵母细胞相比,FAS等基因表达量呈在胚胎发育各个时期呈上升趋势,ACADM等基因表达量呈下降趋势.本研究为深入了解脂肪酸代谢在早期胚胎发育过程中的作用机制奠定了基础.As an energy source,lipids play an important role in oocyte maturation,embryonic development and stem cell proliferation.There are abundant lipids in porcine oocytes and early embryos compared to other cells,but the mechanism of fatty acid metabolism during porcine early embryonic development is poorly understood.In this study,the oocyte maturation medium which contain10% porcine follicular fluid was used.By comparing the cleavage and blastocyst rate of parthenogenetic embryos derived from electro-activation only,electro-activation plus CB or 9% ethanol for3 min,8 min and 11 min,respectively,it was confirmed that the electro-activation method was the best method to obtain parthenogenetic embryos(81.69%±0.41% of cleavage rate and 26.45%±0.28%of blastocyst rate).Immunofluorescence staining showed that the cell number of parthenogenetic blastocysts was significantly lower than in vitro fertilized blastocysts.The lipid droplets were found located around the nucleus according to BODIPY 493/503 staining.However,H2 Av was only found in the nucleus but not in lipid droplets.Moreover,the lipid droplet content in morula was the highest when compared with other developmental stages,and the diameter of lipid droplet reached the maximum in the 8-cell stage.RT-PCR results showed that the expression of FAS and other genes increased in early embryos compared to oocytes.The expression of ACADM and other genes decreased.This study laid a foundation for further understanding the mechanism of fatty acid metabolism in porcine early embryonic development.

关 键 词：猪 孤雌胚胎 脂滴 

分 类 号：S814.6[农业科学—畜牧学] Q95[农业科学—畜牧兽医]