摘要
本试验旨在探究添加低密度脂蛋白(low density lipoprotein,LDL)对玻璃化冷冻解冻后MⅡ期猪卵母细胞发育率、线粒体膜电位及透明带泛素化水平的影响。采用线粒体膜电位特异性探针JC-1检测各处理组线粒体膜电位,并采用SDS-PAGE及Western blotting方法分析不同处理组MⅡ期猪卵母细胞透明带蛋白泛素化水平。结果显示,玻璃化冷冻法处理中,10mg/mL LDL处理组MⅡ期卵母细胞正常率和成熟率(71.92%和69.86%)显著高于对照组(58.26%和54.55%)(P<0.05),最接近未冷冻组。10mg/mL LDL处理组MⅡ期卵母细胞线粒体膜电位显著高于对照组、1和20mg/mL LDL组(P<0.05)。免疫印迹结果显示,未冷冻组和LDL处理组MⅡ期卵母细胞ZP1、ZP2及ZP3蛋白分别在61、80、106ku发生不同程度的泛素化标记;10 mg/mL LDL处理组ZPs(ZP1、ZP2、ZP3)蛋白泛素化水平显著高于1和20 mg/mL LDL组(P<0.05),但显著低于非冷冻组(P<0.05)。结果表明LDL可改善玻璃化冷冻解冻培养后MⅡ期猪卵母细胞成熟率、线粒体膜电位及透明带泛素化水平。
The purpose of this study was to determine the effect of LDL addition on oocyte developmental rate,mitochondrial membrane potential and ZP protein ubiquitination level in MⅡoocytes from vitrifiedwarmed GV oocytes.Mitochondrial membrane potentials were labeled with specific probe JC-1,and ZP protein samples of different treatment groups in MⅡoocytes were analyzed by SDS-PAGE and Western blotting methods.The results showed that normal rate and in vitro maturation of MⅡoocytes in the10mg/mL LDL group（71.92% and 69.86%）were significantly higher than those in control group（58.26%and 54.55%）（P〈0.05）,that were nearest to non-vitrified group.The mitochondrial membrane potential of MⅡoocytes in the 10mg/mL LDL group was significantly higher than those in control group and 1,20mg/mL LDL groups（P〈0.05）.The ubiquitinated ZP1,ZP2 and ZP3of MⅡoocytes in non-vitrified group and LDL treated groups were labeled at 61,80 and 106ku,respectively.The ubiquitinated ZPs（ZP1,ZP2 and ZP3）level of MⅡoocytes in the 10mg/mL LDL group was significantly higher than those in 1and 20mg/mL LDL groups（P〈0.05）,while significantly lower than that of non-vitrified group（P〈0.05）.In conlusion,LDL could improve the maturation rate,mitochondrial membrane potential and ubiquitinated ZPs level of MⅡoocytes from vitrified-warmed GV oocytes.
出处
《中国畜牧兽医》
CAS
北大核心
2017年第7期1961-1966,共6页
China Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金资助项目(31260529)
吉林省科技厅重点科技攻关项目(20150204079NY)
关键词
卵母细胞
玻璃化冷冻
低密度脂蛋白
线粒体膜电位
透明带泛素化
oocytes
vitrification
low density lipoprotein（LDL）
mitochondrial membrane potential
ZP protein ubiquitination
