摘要
本研究旨在探讨3种冷冻方法对猪MII期卵母细胞玻璃化冷冻后线粒体的分布和超微结构变化的影响。通过透射电镜、Rhodamine-123(R-123)荧光染色和体外发育观察,结果显示,(1)无论是FDA-DAPI复染存活率,还是孤雌激活卵裂率,CLV(Cryoloop vitrification)法(72.00%,7.22%)效果最好,OPS(Open Pulled Straw)法(60.00%,4.85%)次之,straw法(42.22%,0%)最低;(2)CLV法经R-123染色后,卵母细胞线粒体的正常分布率(52.24%)要比其它2组要高(OPS,48.65%;straw,37.68%),但三者之间没有显著差异(P>0.05);(3)透射电镜超微结构表明,冻后卵母细胞线粒体变得粗糙和模糊,有的线粒体嵴减少甚至消失。结果表明,冷冻过程中卵母细胞线粒体的分布和形态损伤严重,CLV法可提高冷冻速率,增强冻后卵母细胞的发育能力,降低冷冻造成的卵母细胞线粒体异常分布比例。
The purpose of this study was to detect the mitochondria distribution and ultrastructure changes that resulted from vitrified porcine MⅡ-stage oocytes with three vitrification techniques. Through transmission electron microscopy （TEM）, R-123 （Rhodamine-123） staining and in vitro development, mitochondria distribution and ultrastructure were observed. The result showed that： （1） Given the survival rate of FDA （fluorescein diacetate）-DAPI （4r,6r-diamidino-2-pheny- lindole dihydrochloride） staining and the cleavage rate after parthenogenetic activation, CLV （Cryoloop vitrification） method （72. 00%, 7. 22%） got the best results. OPS （Open Pulled Straw,60.00%, 4.85%） and straw （42.22%, 0%） methods were followed by; （2） The rate of normal mitochondria distribution in CLV group （52.24%） was higher than that in the two others groups （OPS, 48.65%‘ straw, 37.68%） , but there was no significant differences among them （P〉0.05） ; （3） Vague mitochondria in thawed oocytes were observed in the ultrastructure of vitrifled oocytes from TEM, and some mitoehondrial ridges were reduced or even disappeared. The result indicated that mitochondria was damaged greatly in the freezing process and CLA method could improve the freezing speed and minimize such damages.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2012年第10期1525-1530,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
上海市科委农业科技成果转化项目(103919N1800)
上海市科技兴农推广项目(2007-3-7)
上海市农科院青年科技发展基金(2011-07)
国家转基因生物新品种培育科技重大专项(2008ZX08006-005
2009ZX08006-014B)
