Hendricks K E M, Hansen P J
Department of Animal Sciences, University of Florida, Gainesville, FL 32611-0910 USA.
Aust Vet J. 2010 Aug;88(8):307-10. doi: 10.1111/j.1751-0813.2010.00585.x.
To determine whether oxidative damage of ejaculated frozen-thawed sperm prior to oocyte insemination in vitro affects the competence of the resultant embryo to develop to the blastocyst stage.
Extended frozen semen from bulls was thawed, subjected to Percoll gradient purification to obtain motile spermatozoa and mixed with medium containing the pro-oxidants menadione or tert-butyl hydroperoxide. After 3 h at 38.5 degrees C, the sperm were washed and used to inseminate oocytes in vitro. Embryo development proceeded until 8 days after insemination.
Treatment of sperm with 15 or 30 micromol/L menadione reduced the proportions of oocytes that cleaved and those that developed to the blastocyst stage; 30 micromol/L menadione reduced the proportion of cleaved embryos that developed to the blastocyst stage at day 8 after insemination. Oocytes inseminated with sperm treated with 150 or 300 micromol/L tert-butyl hydroperoxide had lower proportions of cleavage and blastocyst development, and the proportion of cleaved embryos becoming blastocysts was also reduced.
Oxidative damage to ejaculated sperm can compromise the ability of the sperm to cause oocyte cleavage and leads to formation of embryos with reduced competence for development.
确定体外卵母细胞授精前冷冻解冻的射出精子的氧化损伤是否会影响所得胚胎发育至囊胚期的能力。
将公牛的冷冻精液解冻，经Percoll梯度纯化以获得活动精子，并与含有促氧化剂甲萘醌或叔丁基过氧化氢的培养基混合。在38.5℃下孵育3小时后，洗涤精子并用于体外授精卵母细胞。胚胎发育持续至授精后8天。
用15或30μmol/L甲萘醌处理精子会降低卵母细胞的分裂比例以及发育至囊胚期的比例；30μmol/L甲萘醌降低了授精后第8天分裂胚胎发育至囊胚期的比例。用150或300μmol/L叔丁基过氧化氢处理的精子授精的卵母细胞，其分裂和囊胚发育的比例较低，并且分裂胚胎形成囊胚的比例也降低。
射出精子的氧化损伤会损害精子导致卵母细胞分裂的能力，并导致发育能力降低的胚胎形成。
